Discuss the potential role of genetic counseling in managing inherited cardiac conditions and its limitations.

Genetic counseling is vital in managing inherited cardiac conditions by educating about genetic risks, organizing screenings, and discussing genetic testing and treatment options. It aims to alleviate psychological impacts of uncertainty and familial risk. However, limitations include the complexity of emotional communication and providing effective preventive strategies, especially for unpredictable conditions like sudden cardiac death .

What challenges are associated with using SNP testing in understanding complex traits, and how might they be addressed?

Testing single SNPs identifies loci with significant associations, but these explain limited phenotypic variance. Challenges include identifying causal variants, understanding the impact of rare variants, SNP-SNP and SNP-environment interactions, and improving algorithms to handle large, noisy datasets. Addressing these may involve leveraging large-scale data integration and developing more comprehensive computational models .

Evaluate the contributions of conserved non-coding elements (CNEs) to gene desert regions and their functional implications.

CNEs prevalent in gene deserts are crucial for regulating distant, significant developmental genes through transcriptional control. They enable complex expression patterns necessary for developmental processes. Alterations in their cis-regulatory functions can result in specific phenotypes linked to malformations .

What role do epigenetic regulators play in the transition from pluripotent ESCs dealing with neuronal differentiation?

Epigenetic regulators modulate gene expression to maintain ESCs' pluripotency and facilitate differentiation. Transitions involve increased repressive regulator expression and decreased activators, with altered microRNA targeting early differentiation genes. These changes sustain ESC epigenotypes while allowing differentiation capacities .

In what ways are chromatin regulators crucial for human disease study and what insights have been gained from model organisms?

Chromatin regulators are central to transcriptional control and disease mechanisms. Studies in model organisms like C. elegans help unravel chromatin complexes' functions, providing insights into genome-wide target regulation in multicellular organisms. This knowledge is crucial for understanding human diseases linked to chromatin dysregulation .

What is the significance of conserved non-coding elements (CNEs) in relation to congenital malformations?

CNEs, acting as gene expression regulators, are critical in proper developmental signaling. Mutations in these regions can alter gene expression without affecting coding regions, leading to dosage-dependent malformations such as brachydactyly, syndactyly, and others. This underscores the importance of regulatory elements in congenital anomalies .

How has the understanding of the genetic basis of cardiomyopathies advanced clinical practice?

Advancements in the molecular genetics of cardiomyopathies, such as identifying desmosomal gene mutations, have improved clinical management by enabling genetic risk assessment, guiding cardiac screenings, and informing family members. This integration into clinical practice enhances patient care and informs therapeutic strategies .

How do epigenetic maps of chromatin provide insights into the regulation of adipogenesis?

Genome-wide chromatin state maps and PPARγ/CTCF localization maps reveal dynamic chromatin remodeling during adipogenesis, identifying pre-adipocyte and adipocyte-specific cis-regulatory elements. Variations in the location of these elements between models, driven by evolutionary transcription factor motif turnover, imply multiple distal regulatory elements' importance in adipogenesis-dependent loci .

How can genetic counseling and testing impact the lives of patients with hereditary conditions and their families?

Genetic counseling helps by providing risk assessments, educating about hereditary conditions, and guiding testing and management options. It supports coping mechanisms within families, helping them navigate uncertainties and formulate preventive and therapeutic strategies .

What role do conserved non-coding elements (CNEs) play in gene regulation, and what are the implications of mutations in these elements?

CNEs serve as essential regulators of gene expression, often located far from the genes they regulate. They promote gene regulation through transcription factor binding and subsequent loop formation between the CNE and the gene's promoter. Mutations affecting the regulatory function of CNEs can result in altered gene expression, contributing to conditions such as brachydactyly and Laurin-Sandrow syndrome due to disrupted dosage-dependent signaling .

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ESHG2010 Abstracts

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ESHG2010 Abstracts

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Volume 18 Supplement 1

June 2010

www.nature.com/ejhg

European Human Genetics Conference 2010 June 12 15, 2010 Gothenburg, Sweden Abstracts

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For In Vitro Diagnostic Use Only. The 3500 Dx and 3500xL Dx Genetic Analyzers and system accessories meet the requirements of the In Vitro Diagnostic Medical Devices Directive (98/79/EC). Fragment analysis is for research use only, and is not intended for human or animal diagnostic or therapeutic procedures. The 3500 Dx Series Systems are for distribution and use in selected countries only. Not for sale in the United States of America. 10-LFT-003 IVD Efficiency Xpert Ad_FINAL.indd 1 All rights reserved. The trademarks mentioned herein are the property of Life Technologies Corporation or their respective owners. 4/29/10 5:13 2010 Life Technologies Corporation.

European Society of Human Genetics

EUROPEAN HUmAN GENEtics cONFERENcE 2010

in conjunction with the

European meeting on Psychosocial Aspects of Genetics

swedish Exhibition & congress centre Gothenburg, sweden saturday, June 12 tuesday, June 15, 2010

Abstracts
www.eshg.org/eshg2010

Committees Board Organisation

European society of Human Genetics ESHG Office European Society of Human Genetics Ms. Karin Knob c/o Vienna Medical Academy Alserstrasse 4 1090 Vienna Austria Tel: +43 1 405 13 83 20 Fax: +43 1 405 13 83 23 Email: [email protected] Website: www.eshg.org Executive Board 2009-2010 President Dian Donnai, UK Vice-President Jean-Jacques Cassiman, BE President-Elect Milan Macek Jr., CZ Secretary-general Helena Kriinen, FI Secretary-general-elect Gunnar Houge, NO Treasurer Andrew Read, UK Observer Jerome del Picchia, AT Scientific Programme Committee Co-Chairs Brunhilde Wirth; DE Han Brunner, NL Members Kristiina Aittomki, FI Corinne Antignac, FR The-Hung Bui, SE Niklas Dahl, SE, Local Host Manolis Dermitzakis, CH Peter Heutink, NL Gunnar Houge, NO Thomas Jensen, DK Helena Kriinen, FI Batsheva Kerem, IL Mark McCarthy, UK Carla Oliveria, PT Olaf Riess, DE Mariano Rocchi, IT Pete Scambler, UK Michael Speicher, AT Eduardo Tizzano, ES Draga Toncheva, BG Mikka Vikkula, BE Cisca Wijmenga, NL Liaison members Segolne Aym, FR Jacqueline Schoumans, CH Han Brunner, NL Brunhilde Wirth, DE Martina Cornel, NL Peter Farndon, UK Ros Hastings, UK Ulf Kristoffersson, SE Heather Skirton, UK

Board members Karen B. Avraham, IL Agnes Bloch-Zupan, FR John Burn, UK Anne Cambon-Thomsen, FR Francoise Clerget-Darpoux, FR Domenico Coviello, IT Gerry Evers-Kiebooms; BE Peter Heutink, NL Klaus W. Kjaer; DK Vaidutis Kuinskas; LT

Jan Lubinski, PL Giuseppe Novelli; IT Tayfun Ozcelik, TR Borut Peterlin, SI Franco Pignatti, IT Alexandre Reymond, CH Jorge Sequeiros, PT Maria Soller, SE Silke Sperling, DE

Further information on structure and organisation can be found on the website www.eshg.org

Future European Human Genetics conferences

European Human Genetics conference 2011 May 28 - 31, 2011 Amsterdam, The Netherlands European Human Genetics conference 2012 June 2 - 5, 2012 Nrnberg, Germany

Table of Contents

EsHG spoken Presentations Plenary Lectures.............................................................................................................................PL1.1 PL5.1 ........................... 4 Concurrent Symposia .....................................................................................................................S01.1 S15.3 .......................... 7 Educational Sessions .....................................................................................................................ES1.1 ES8.2 ....................... 16 Concurrent Sessions ......................................................................................................................C01.1 C16.6 ....................... 18 EsHG Posters P01. Genetic counseling, including Psychosocial aspects, Genetics education, Genetic services, and Public policy ................................................................................................P01.01 P01.68 .................... 43 P02. Clinical genetics and Dysmorphology ....................................................................................P02.001 P02.203 ................ 58 P03. Cytogenetics ..........................................................................................................................P03.001 P03.136 .............. 103 P04. Reproductive genetics............................................................................................................P04.01 P04.44 .................. 134 P05. Prenatal and perinatal genetics..............................................................................................P05.01 P05.70 .................. 144 P06. Cancer genetics .....................................................................................................................P06.001 P06.158 .............. 160 P07. Cancer cytogenetics...............................................................................................................P07.01 P07.27 .................. 195 P08. Statistical genetics, includes Mapping, linkage and association methods .............................P08.01 P08.58 .................. 201 P09. Complex traits and polygenic disorders .................................................................................P09.001 P09.142 .............. 215 P10. Evolutionary and population genetics, and Genetic epidemiology .........................................P10.01 P10.64 .................. 252 P11. Genomics, Genomic technology including bioinformatics methods, gene structure and gene product function and Epigenetics. ...................................................................P11.001 P11.142 .............. 269 P12. Molecular basis of Mendelian disorders .................................................................................P12.001 P12.223 .............. 300 P13. Metabolic disorders ................................................................................................................P13.01 P13.57 .................. 350 P14. Therapy for genetic disorders ................................................................................................P14.01 P14.18 .................. 362 P15. Laboratory and quality management......................................................................................P15.01 P15.16 .................. 366 EmPAG spoken Presentations Plenary Lectures.............................................................................................................................EPL1.1 EPL 7.5 ................ 371 Concurrent Sessions ......................................................................................................................EPC5.1 EPC5.6 ................ 380 Workshops......................................................................................................................................EWS1.1 EWS3.2 .............. 382 EmPAG Posters EP01. Reproductive issues in genetics, prenatal and preimplantation genetic diagnosis ..............EP01.01 EP01.05 ............. 383 EP02. Prenatal and newborn screening .........................................................................................EP02.01 EP02.06 ............. 384 EP03. Risk perception and genetic testing .....................................................................................EP03.01 EP03.03 ............. 385 EP04. Access to genetic services (challenges in Europe)..............................................................EP04.01 EP04.04 ............. 386 EP05. Lay beliefs and public understanding of genetics ................................................................EP05.01 EP05.03 ............. 387 EP06. Predictive testing: process and impact ................................................................................EP06.01 EP06.03 ............. 387 EP07. Psycho-social issues in cancer genetics .............................................................................EP07.01 EP07.06 ............. 388 EP08. Psychosocial issues in cardiac genetics ..............................................................................EP08.01 EP08.02 ............. 389 EP09. Predisposition to common diseases: genetic testing and preventive behaviour .................EP09.01 EP09.02 ............. 390 EP10. Genetic counselling: communicating genetic information ....................................................EP10.01 EP10.10 ............. 390 EP11. Strategies to facilitate decision making in genetics..............................................................EP11.01 EP11.02 ............. 393 EP12. Family dynamics and genetic conditions .............................................................................EP12.01 EP12.07 ............. 393 EP13. Living with genetic disease ..................................................................................................EP13.01 EP13.05 ............. 395 EP14. Evaluation of psycho-social interventions in genetics..........................................................EP14.01 ............................... 396 EP15. Other relevant psychological and social topics in genetics..................................................EP15.01 EP15.10 ............. 396 Author Index ................................................................................................................................................................................ 399 Keyword Index ............................................................................................................................................................................. 443

Abstracts marked with * are talks of Young investigator Candidates Abstracts marked with ** are ESHG Poster Award Candidates

Plenary Lectures Abstracts of EsHG Plenary sessions

PL1.1 Peopling of the New World high-Arctic: A Genetic Perspective PL1.4 Genetics of cancer predisposition
L. Aaltonen; University of Helsinki, Molecular and Cancer Biology Program, Helsinki, Finland.

M. Raghavan1, E. Willerslev2; 1 Centre for GeoGenetics, Natural History Museum of Denmark, University of Copenhagen, Copenhagen, Denmark, 2Centre for GeoGenetics,Natural History Museum of Denmark, Copenhagen, Denmark.

No abstract reiceved as per date of printing, please check the abstract section on the conference website www.eshg.org/eshg2010 for possible updates. PL2.1* the Effect of translocation-induced Nuclear Reorganization on Gene Expression

Distinct cultural waves swept through the New World high-Arctic (Canada and Greenland), leaving behind well-preserved material and biological traces in the permafrost. This talk will focus on a major paleogenetic endeavor aimed at determining the genetic signatures of the three ancient high-Arctic cultures - Saqqaq, Dorset and Thule and ascertaining any genetic relationships between them by analyzing bone, hair and teeth samples from individuals excavated from sites across the Canadian Arctic and Greenland. Current work constitutes the use of state-of-the-art high throughput sequencing to identify genome-wide markers that would help resolve the phylogenetic relationships of the Saqqaq, Dorset and Thule with respect to each other as well as to modern Inuit and Native American populations. Results from this analysis will help disentangle issues surrounding the origins of the first humans in the region, the timing of these migrations, and provide some perspective on the extent to which they have individually contributed to the genetic history of the New World Arctic. PL1.2 monogenic diabetes: the success of molecular genetics for improved diagnosis and treatment
P. Njlstad; Department of Clinical Medicine, University of Bergen, and Department of Pediatrics, Haukeland University Hospital, Bergen, Norway.

L. Harewood1, F. Schtz1,2,3, S. Boyle4, P. Perry4, M. Delorenzi3,2, W. A. Bickmore4, A. Reymond1; 1 Center for Integrative Genomics, Lausanne, Switzerland, 2Swiss Institute of Bioinformatics (SIB), Lausanne, Switzerland, 3National Center of Competence in Research (NCCR) Molecular Oncology, Lausanne, Switzerland, 4MRC Human Genetics Unit, Institute of Genetics and Molecular Medicine, Edinburgh, United Kingdom.

Monogenic diabetes results from mutation in a single gene primarily affecting pancreatic beta or acinar cell function. The prevalence is 1-2 % of all diabetes. Monogenic diabetes is, however, frequently misdiagnosed as type 1 or type 2 diabetes. Knowledge of the genetic etiology of monogenic diabetes has proved essential for improved treatment, prediction of prognosis, genetic counseling and identification and screening of additional family members. Monogenic diabetes can be divided in two main groups; neonatal diabetes and maturity-onset diabetes of the young (MODY). Neonatal diabetes is commonly used when diabetes occurs before the age of six months. Mutations in several beta cell genes can cause neonatal diabetes, the most important being KCNJ11 and ABCC8 that encode the Kir6.2 and SUR1 subunits of the ATP-sensitive potassium channel, respectively. Children having a mutation in either of these genes can be treated with sulfonylurea rather than insulin injections and with better glycemic control. Some ten genes can cause MODY. It is important to diagnose GCKMODY (MODY2) since this is a mild form for diabetes that seldom requires treatment and in which late-diabetes complications are rare. Two other common forms are due to mutations in the transcription factor genes HNF4A (MODY1) and HNF1A (MODY3). These are clinically nearly indistinguishable and are commonly misdiagnosed as type 1 or type 2 diabetes. Sulfonylurea sensitivity is retained why these forms can successfully be treated with low doses of sulfonylureas. Subjects with mutations in HNF1B (MODY5) or CEL (MODY8) often develop both exocrine and endocrine dysfunction, and hence require both pancreatic enzyme supplement and insulin. Monogenic diabetes should be suspected in subjects with diabetes and an unusual presentation or development. Most will have a positive family history of diabetes, a primary beta cell dysfunction and be negative for antibodies associated with type 1 diabetes. PL1.3 A human protein atlas to study human genetics

Translocations are known to affect expression of genes at the breakpoints and, in the case of unbalanced translocations, alter gene copy number. However, a comprehensive understanding of the functional impact of this class of variation is lacking. We have studied the effect of balanced chromosomal rearrangements on gene expression by comparing transcriptomes of cell lines from controls and individuals with the t(11;22)(q23;q11) translocation. The number of differentially expressed transcripts between translocation carrying and control cohorts is significantly higher than that observed between control samples alone, suggesting that balanced rearrangements have a greater effect on gene expression than normal variation. Many of the affected genes are located on the derivative chromosome 11. We show that this chromosome is concomitantly altered in its spatial organization, occupying a more central position in the nucleus than its non-rearranged counterpart. Consistently, we observe nuclear repositioning of genes that show differential expression in balanced translocation carriers as compared to controls. The movement of the derivative 11 also results in nuclear repositioning of other, non-translocated chromosomes. Our results are consistent with recent studies that experimentally altered nuclear organization and indicate that nuclear position plays a functional role in regulating the expression of some genes. Our study suggests that chromosomal translocations can result in hitherto unforeseen, large-scale changes in gene expression that are the consequence of alterations in normal chromosome territory positioning. This has implications for the patterns of gene expression change seen during tumorigenesis associated genome instability and during karyotype changes that lead to speciation. PL2.2* PDZD7 is a modifier of and digenic contributor to retinal disease in Usher syndrome

H. J. Bolz1,2, I. Ebermann1, J. B. Phillips3, M. C. Liebau4, R. K. Koenekoop5, B. Schermer4,6, I. Lopez5, E. Schfer7, A. F. Roux8,9, C. Dafinger1, A. Bernd10, E. Zrenner10, M. Claustres8,9, B. Blanco3, G. Nrnberg11, P. Nrnberg11,6, R. Ruland1, M. Westerfield3, T. Benzing4,6; 1 Institute of Human Genetics, University Hospital of Cologne, Cologne, Germany, 2Bioscientia Center for Human Genetics, Ingelheim, Germany, 3 Institute of Neuroscience, University of Oregon, Eugene, OR, United States, 4 Department of Medicine and Centre for Molecular Medicine, University Hospital of Cologne, Cologne, Germany, 5McGill Ocular Genetics Laboratory, McGill University Health Centre Research Institute, Montreal, QC, Canada, 6 Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases, University of Cologne, Cologne, Germany, 7Praxis fr Humangenetik, Hamburg, Germany, 8CHU Montpellier, Laboratoire de Gntique Molculaire, Montpellier, France, 9Inserm, U827, Montpellier, France, 10Centre for Ophthalmology, Institute for Ophthalmic Research, University of Tbingen, Tbingen, Germany, 11Cologne Center for Genomics and Institute for Genetics, Cologne, Germany.

M. Uhln; KTH Biotechnology, AlbaNova University Center, Stockholm, Sweden.

No abstract reiceved as per date of printing, please check the abstract section on the conference website www.eshg.org/eshg2010 for possible updates.

Usher syndrome is a genetically heterogeneous recessive disease with hearing loss and retinitis pigmentosa (RP). It frequently presents with unexplained, often intrafamilial, variability of the visual phenotype. Here, we identify PDZD7, encoding a homolog of proteins mutated in Usher subtypes 1C and 2D (USH1C, USH2D). We demonstrate interaction between PDZD7 and two Usher disease proteins, GPR98 (USH2C) and USH2A, and their colocalization in the photoreceptors connecting cilium region. On a homozygous USH2A mutation background, PDZD7 aggravates RP. Heterozygous PDZD7 mutations

Plenary Lectures
were present with truncating mutations in USH2A, GPR98, and an unidentified locus. We validated the human genotypes using zebrafish, because visual defects are typically not evident in mouse models of Usher syndrome. Our findings in zebrafish were consistent with digenic inheritance of PDZD7 and GPR98, and with PDZD7 as a retinal disease modifier in USH2A patients. Pdzd7 knockdown produced an Usher-like phenotype in zebrafish, exacerbated retinal cell death in combination with ush2a or gpr98, and significantly reduced Gpr98 localization in the region of the photoreceptor connecting cilium. Our data challenge the view of Usher syndrome as a traditional Mendelian disorder. As in Bardet-Biedl syndrome, reclassification of Usher syndrome as an oligogenic disease permits a better understanding of its phenotypic variability. PL2.3* The identification of 180 genetic loci involved in adult height variation highlights the complex genetic architecture of polygenic traits
4

Department of Medical Genetics, Hospital Peditrico de Coimbra, Coimbra, Portugal, 5Centre for Human Genetics, Cliniques Universitaires St Luc, Universit Catholique de Louvain, Brussels, Belgium, 6Central and Southern Regional Genetics Services, Wellington Hospital, Wellington, New Zealand, 7 Centro de Gentica Mdica Doutor Jacinto Magalhes/INSA, Porto, Portugal, 8 Department of Medical Genetics, Sydney Childrens Hospital, Sydney, Australia, 9Department of Clinical Genetics, The Childrens Hospital at Westmead, Sydney, Australia, 10Clinical Genetics Unit, Birmingham Womens Hospital, Birmingham, United Kingdom, 11Institute of Human Genetics, Newcastle upon Tyne Hospitals NHS Trust, Newcastle upon Tyne, United Kingdom, 12Northern Regional Genetics Service, Auckland, New Zealand, 13 SA Clinical Genetics Service, Womens & Childrens Hospital site, Adelaide, Australia.

K. Estrada1, G. Lettre2, H. Lango3, S. I. Berndt4, M. N. Weedon3, G. R. Abecasis5, M. Boehnke6, C. Gieger7, D. Gudbjartsson8, N. L. Heard-Costa9, A. U. Jackson6, M. I. McCarthy10, A. Smith11, N. Soranzo12, A. G. Uitterlinden1, F. Rivadeneira1, T. M. Frayling3, J. N. Hirschhorn13; 1 Department of Internal Medicine, Erasmus University MC, Rotterdam, Netherlands, 2Montreal Heart Institute (Research Center), Universit de Montral, Montral, QC, Canada, 3Genetics of Complex Traits, Peninsula Medical School, Exeter, United Kingdom, 4Division of Cancer Epidemiology and Genetics, National Cancer Institute, National Institutes of Health, Department of Health and Human Services, Bethesda, MD, United States, 5Center for Statistical Genetics, Department of Biostatistics, University of Michigan, Ann Arbor, MI, United States, 6Department of Biostatistics, School of Public Health, University of Michigan, Ann Arbor, MI, United States, 7Institute of Epidemiology, Helmholtz Zentrum Mnchen, Neuherberg, Germany, 8deCODE genetics, Reykjavik, Iceland, 9Department of Neurology, Boston University School of Medicine, Boston, MA, United States, 10Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford, United Kingdom, 11Icelandic Heart Association, Kopavogur, Iceland, 12Wellcome Trust Sanger Institute, Hinxton, United Kingdom, 13Program in Medical and Population Genetics, Broad Institute of Harvard and Massachusetts Institute of Technology, Boston, MA, United States.

Height is a classic polygenic trait: it has been proposed for nearly 100 years that variants in multiple genes influence height; indeed up to 90% of variation in height is attributable to inherited variation. As part of the Genetic Investigation of ANthropometric Traits (GIANT) Consortium, we performed a meta-analysis of genome-wide association studies of adult height, encompassing 2.8 million single nucleotide polymorphisms (SNPs) and 133,800 individuals of European ancestry from 50 individual studies. We detected 207 distinct loci with a SNP associated with height at P<5x10-6 (versus 5 expected by chance). When we examined the 207 SNPs in a replication dataset of over 51,000 samples, 180 reached a combined P<5x10-8. Many loci have multiple associated variants: conditional analysis identified 19 loci that have additional independent strong associations. Together, the associated variants explain up to 12% of height variation. Using a polygenic model, the explained variance reached a maximum value of 16% when including signals associated at P < 5x10-3. The associated loci are strongly enriched for genes known to be mutated in monogenic disorders of skeletal growth (P<0.001), and implicate genes in relevant biological pathways, including hedgehog signaling, TGF-beta signaling, extracellular matrix, and growth hormone signaling. In conclusion, we have identified nearly 200 genetic loci influencing adult height. These loci are enriched for genes in biological pathways relevant to human growth, and support a notion that allelic heterogeneity may be a common feature of polygenic traits. PL2.4* De novo mutations of SETBP1 cause schinzel-Giedion syndrome

Schinzel-Giedion syndrome is characterized by severe mental retardation and characteristic facial features. Most patients die before the age of 10 years. Almost all patients occur sporadically raising the possibility that de novo dominant mutations could be the underlying mechanism. The exomes of four affected individuals with SchinzelGiedion syndrome were sequenced to a mean coverage of 43-fold. On average 21,800 genetic variants were identified per patient. After filtering against known variants and selecting variants affecting the same gene in all patients, only a single candidate gene remained, SETBP1 which encodes SET binding protein 1. Sanger sequencing confirmed the presence of SETBP1 mutations in these 4 individuals as well as in 8 additional Schinzel-Giedion syndrome patients. All mutations occurred de novo, and two mutations were recurrent: c.2602G>A and c.2612T>C in 4 and 5 patients, respectively. Because all mutations were missense, and clustered to a ultra high conserved 11bp exonic region of SETBP1, it is highly likely that they cause disease through either a dominant negative or a gain-of-function effect. In conclusion, our study shows the potential of exome sequencing for disease gene identification by unraveling the first dominant Mendelian disorder. It is highly likely that the mutations cause disease through either a dominant negative or a gain-of-function effect. Exome sequencing is particularly useful for identifying these types of mutations for which no other genomewide approach is applicable. De novo mutations may be a frequent cause of frequent sporadic conditions with reduced fecundity such as congenital malformations, mental retardation, and psychiatric disorders. PL2.5 complete genome sequencing and analysis of diploid African-American and mexican-American genomes: implications for personal ancestry reconstruction and multi-ethnic medical genomics
F. M. De La Vega1, J. D. Degehnardt2, S. Musharoff2, K. Bryc2, J. M. Kidd3, V. Seth4, S. Stanley1, C. Monighetti1, E. Levandowsky4, M. Barker1, A. Brisbin2, A. Keinan2, A. Clark2, C. D. Bustamante3,2; 1 Life Technologies, Foster City, CA, United States, 2Cornell University, Ithaca, NY, United States, 3Stanford University, Stanford, CA, United States, 4Life Technologies, Beverly, MA, United States.

A. Hoischen1, B. van Bon1, C. Gilissen1, P. Arts1, B. van Lier1, M. Steehouwer1, P. de Vries1, R. de Reuver1, N. Wieskamp1, G. Mortier2, K. Devriendt3, M. Z. Amorim4, N. Revencu5, A. Kidd6, M. Barbosa7, A. Turner8, J. Smith9, C. Oley10, A. Henderson11, I. M. Hayes12, E. Thompson13, H. G. Brunner1, B. B. A. de Vries1, J. A. Veltman1; 1 Radboud University Nijmegen Medical Centre, Nijmegen, Netherlands, 2 Centre for Medical Genetics, University Hospital Antwerp, Antwerp, Belgium, 3 Centre for Clinical Genetics, Leuven University Hospital, Leuven, Belgium,

Understanding the contribution of rare and common genetic variants to disease susceptibility will likely require multi- and trans-ethnic sequencing studies that compare the genomes of many individuals with and without a particular disease. Of particular importance will be accounting for the role of population stratification at fine scales both in terms of genomic and geographic location. Here, we present results from sequencing, assembly, and genomic analysis of two diploid genomes from Phase 3 HapMap sequenced to ~20X coverage using the SOLiD System. The donor individuals are of Mexican and African ancestry and represent the first admixed genomes to be sequenced to high coverage. We demonstrate that genomic sequencing provides finer resolution of admixture breakpoints based on allele frequency estimates from HapMap and the 1000 Genomes Project. For each admixed genome, we use the distribution of admixture breakpoints to infer the personal admixture history of the sample and patterns of genomic diversity to reconstruct the demographic history of European, African, and Native American continental populations. Furthermore, we compare the distribution of functional and putatively neutral genetic variation among 12 sequenced genomes and find that difference in demographic history may account for statistically significant, differences in distributions of synonymous vs. benign, possibly damaging, and probably damaging non-synonymous coding variants. Finally, we use the SOLiD comparative personal genomic data sets and 1000 Genomes

Plenary Lectures
data to quantify the relative proportions of private, rare, and common functional and neutral genetic within and among populations. PL2.6 A mutation in the 3UtR of the HDAc6 gene abolishing the post-transcriptional regulation mediated by hsa-miR-433 is linked to a new form of dominant X-linked chondrodysplasia.

directly regulated by FOXP2. Intriguingly, we found that CNTNAP2 is itself associated with common cases of language impairment; this target has also been implicated in language delays of autistic children. High-throughput screening has enabled us to isolate additional putative targets of FOXP2, including multiple genes involved in neurite outgrowth and synaptic plasticity. Moving to animal models of FOXP2 dysfunction, we have shown that point mutations implicated in human speech deficits yield impaired motor-skill learning in mutant mice. Electrophysiological recording suggests that this may be mediated by altered plasticity of Foxp2-expressing circuitry. Together with findings from other model systems, these data indicate that the contributions of FOXP2 to human speech and language are built on evolutionarily ancient roles in neural circuits involved in sensorimotor integration and motor-skill learning. Overall, this work demonstrates how we can begin to bridge gaps between molecules, neurons and the brain, helping us to build more sophisticated models of the relationships between genes, speech and language. PL3.2 mice, chimpanzees and the molecular basis of speech
W. Enard; Max-Planck-Institute of Evolutionary Anthropology, Leipzig, Germany.

B. Arveiler1,2, D. Simon1, M. Barillot1, B. Laloo3, T. Barnetche1, C. Rooryck1,2, C. Blanchard1, I. Burgelin1, M. Marche1, I. Coupry1, N. Chassaing4,5, B. GilbertDussardier6, C. Grosset3, D. Lacombe1,2; 1 Universit Victor Segalen Bordeaux 2, Bordeaux, France, 2CHU de Bordeaux, Bordeaux, France, 3INSERM U889, Bordeaux, France, 4INSERM U563, Toulouse, France, 5CHU de Toulouse, Toulouse, France, 6CHU de Poitiers, Poitiers, France.

A family with dominant X-linked chondrodysplasia was previously described. The disease locus was ascribed to a 24 Mb interval in Xp11.3q13.1. We have identified a variant (c.*281A>T) in the 3UTR of the HDAC6 gene that totally segregates with the disease. The variant is located in the seed sequence of hsa-miR-433. Our data showed that, in MG63 osteosarcoma cells, hsa-miR-433 (miR433) down-regulated both the expression of endogenous HDAC6 and that of an eGFP-reporter mRNA bearing the wild-type 3UTR of HDAC6. This effect was totally abrogated when the reporter mRNA bore the mutated HDAC6 3UTR. The HDAC6 protein was found to be over-expressed in thymus from an affected male foetus. Concomitantly, the level of total alphatubulin, a target of HDAC6, was found to be increased in the affected foetal thymus, whereas the level of acetylated alpha-tubulin was found to be profoundly decreased. Skin biopsies were obtained from a female patient who presented a striking body asymmetry with hypotrophy of the left limbs. The mutated HDAC6 allele was expressed in 31% of left arm-derived fibroblasts, whereas it was not expressed in the right arm. Overexpression of HDAC6 was observed in left arm-derived fibroblasts. Altogether these results strongly suggest that this HDAC6 3UTR variant suppressed hsa-miR-433-mediated post-transcriptional regulation causing the overexpression of HDAC6. This variant is likely to constitute the molecular cause of this new form of X-linked chondrodysplasia. This represents to our knowledge the first example of a skeletal disease caused by the loss of a miRNA-mediated post-transcriptional regulation on its target mRNA. PL2.7 the BRcA gene patent dispute: breaking news from America
G. Matthijs; Center for Human Genetics, K.U.Leuven, Leuven, Belgium.

Identifying the genetic changes responsible for the phenotypic differences between humans and their close primate relatives is important from an evolutionary, medical and cultural perspective. The primary challenge facing researchers today, after analyzing the genomic data, is experimentally testing hypotheses concerning the genetic basis for human-specific traits. One of the more prominent hypotheses of this kind states that two amino acid changes in the transcription factor FOXP2 have been fixed in humans by positive selection due to some effect on speech and language. We have introduced these substitutions into the endogenous Foxp2 gene of mice. Although these mice are generally healthy, they have qualitatively different ultrasonic vocalizations, decreased exploratory behavior and decreased dopamine concentrations in the brain suggesting that the humanized Foxp2 allele affects basal ganglia. In the striatum, a part of the basal ganglia affected in humans with a speech deficit due to a non-functional FOXP2 allele, we find that medium spiny neurons have increased dendrite lengths and increased synaptic plasticity. Since mice carrying one non-functional Foxp2 allele show opposite effects, this suggests that alterations in cortico-basal ganglia circuits might have been important for the evolution of speech and language in humans. PL3.3 Evolution of Human Language

No abstract reiceved as per date of printing, please check the abstract section on the conference website www.eshg.org/eshg2010 for possible updates. PL3.1 Neurogenetic pathways regulated by FOXP2, a gene implicated in speech and language
S. E. Fisher; Wellcome Trust Centre for Human Genetics, Oxford, United Kingdom.

W. T. Fitch; University of St Andrews, School of Psychology, St. Andrews, Fife, Scotland, United Kingdom.

No abstract reiceved as per date of printing, please check the abstract section on the conference website www.eshg.org/eshg2010 for possible updates. PL5.1 ESHG Award Lecture : DNA fingerprinting and the turbulent genome

People who carry rare heterozygous mutations disrupting the FOXP2 gene have problems mastering the complex sequences of mouth movements needed for speech, along with deficits in many aspects of expressive and receptive language. The gene encodes a highly conserved transcription factor that helps regulate development and function of neuronal subpopulations in a wide range of non-speaking vertebrates, although evidence suggests that its role(s) may have been modified during human evolution. It is emphasised that FOXP2 is not the mythical gene for speech, but represents one piece of a complex puzzle. I will describe how FOXP2 can be used as a unique window into key neurogenetic pathways via an array of complementary approaches. For example, using functional genomic screening of human neurons grown in the laboratory, we identified the CNTNAP2 gene (a member of the neurexin superfamily) as a downstream target

A. Jeffreys; University of Leicester, Department of Genetics, Leicester, United Kingdom.

The accidental development of DNA fingerprinting 25 years ago marked the birth of DNA-based identification. I will discuss the origins and evolution of DNA testing, the creation of major national DNA databases and the extraordinary impact that DNA has had on forensic and legal medicine. I will also discuss how DNA fingerprinting identified some of the most unstable regions in the human genome, allowing us to study human DNA evolution in real time and explore the fundamental processes of mutation and recombination that are the ultimate source of all human DNA variation.

Concurrent Symposia Abstracts of EsHG concurrent symposia

s01.1 the cradle of constitutional chromosome rearrangements is the cleavage stage embryo
J. Vermeesch; Center for Human Genetics, K.U.Leuven, Leuven, Belgium.

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Genome architectural features consisting of low-copy repeats (LCRs), also called segmental duplications, can stimulate and mediate NAHR. There are positional hotspots for the crossovers within the LCRs. We recently elucidated a DNA replication mechanism for nonrecurrent rearrangements that we termed FoSTeS - Fork Stalling and Template Switching. A newer model, microhomology-mediated break-induced replication or MMBIR, provides further molecular mechanistic details and may be operative in all life forms as a means to process one-ended, double-stranded DNA generated by collapsed forks. Rearrangements introduce variation into our genome for selection to act upon and as such serve an evolutionary function analogous to base pair changes. Genomic rearrangements may cause Mendelian diseases and complex traits such as obesity and neurobehavioral phenotypes. The mechanisms by which rearrangements convey phenotypes are diverse and include gene dosage, position effects, unmasking of coding region mutations (cSNPs) or other functional SNPs, creating gain-offunction fusion genes at the breakpoints, and perhaps through effects of transvection. De novo genomic rearrangements have been shown to cause both chromosomal and Mendelian disease, as well as sporadic traits, but our understanding of the extent to which genomic rearrangements, gene CNV, and/or gene dosage alterations are responsible for common and complex traits remains rudimentary.
1. Hastings, PJ, Ira, G, Lupski, JR (2009) A microhomology-mediated break-induced replication model for the origin of copy number variation. PLoS Genetics 5 :1-9 [e100327]. 2. Lupski, JR (2009) Genomic disorders ten years on. Genome Medicine 1 :42.1 - 42.11. 3. Zhang, F, Gu W, Hurles, ME, Lupski, JR (2009) Copy number variation in health, disease, and evolution. Annual Reviews of Genomics and Human Genetics 19:451-481 4. Zhang, F, Carvalho, CMB, Lupski, JR (2009) Complex human chromosomal and genomic rearrangements. Trends in Genetics 25:298-307. 5. Hastings PJ, Lupski, JR, Rosenberg, SM, Ira, G (2009) Mechanisms of change in gene copy number. Nature Reviews Genetics 10:551-564. 6. Stankiewicz, P. and Lupski, J.R. (2010). Structural variation in the human genome and its role in disease. Annual Reviews of Medicine 61:437-455. 7. Carvalho, C.M.B., Zhang, F., Lupski, J.R. (2010). Evolution in health and medicine. Sackler colloqium; Genomic disorders - a window into human gene and genome evolution. Proc. Natl. Acad. Sci. U.S.A. 107:1765-1771.

We developed several novel tools to genome wide screen for CNVs and SNPs in single cells. When applied to cleavage stage embryos from young fertile couples we discovered, unexpectedly, an extremely high incidence of chromosomal instability, a hallmark of tumorigenesis. Not only mosaicisms for whole chromosome aneuploidies and uniparental disomies but also frequent segmental deletions, duplications and amplifications that were reciprocal in sister blastomeres were detected in most cleavage stage embryos implying the occurrence of breakage-fusion-bridge cycles. In addition, we demonstrate the existence of those rearrangements in interphase nuclei. The type of rearrangements observed can likely explain the majority of constitutional rearrangements seen in miscarriages as well as live births such as deletions, duplications, inverted deletions duplications, ring chromosomes and mosaicisms of all of those rearrangements. The high frequency of chromosomal imbalances in cleavage stage embryos make it likely that chromosomal disorders originate post-zygoticaly. s01.2 How common is somatic mosaicism for DNA copy Number Variations (cNVs)?

J. P. Dumanski; Department of Genetics and Pathology, Uppsala University, Uppsala, Sweden.

DNA Copy Number Variation (CNV) has emerged as the most common form of human inter-individual genetic differences and this is important for basic research in biology/genetics as well as for disease-oriented translational science. We have recently discovered that monozygotic (MZ) twins frequently display within-pair differences in CNV profiles, which indicates the feasibility of studying MZ twins, discordant for established phenotypes in search for disease-causing aberrations. In addition, recent analysis of differentiated human tissues of normal deceased subjects supports the notion that somatic CNV mosaicism is underestimated. We tested multiple tissues from three people for differences in CNV profiles and observed changes, affecting a single organ or one or more tissues of the same person. Our results from MZ twins and CNV differences between normal differentiated human tissues of the same person suggest that humans are commonly affected by mosaicism for stochastic CNVs, which occur in a substantial fraction of normal cells and are detectable by available array-based methods. However, the somatic DNA copy number variation is not well studied. The work in the group focuses on establishment of baseline of somatic CNV (the normal frequency and genomic distribution of somatic CNVs) in phenotypically unselected, healthy, concordant MZ twins and comparisons of different tissues from the same individuals. We also study differences in the CNV distribution and/or frequency in MZ twins discordant for various disease phenotypes in search for new diseaserelated biomarkers. s01.3 Genomic Disorders: mechanisms and assays for cNV associated with neuropsychiatric and other disease traits

s02.1 Ethical issues in large scale genomics research

T. Caulfield; Health Law Institute, Law Centre, University of Alberta, Edmonton, AB, Canada.

J. R. Lupski; Baylor College of Medicine, Department of Molecular and Human Genetics, Houston, TX, United States.

Whereas Watson-Crick DNA base pair changes have long been recognized as a mechanism for mutations, rearrangements of the human genome including deletions, duplications, and inversions have been appreciated only more recently as a significant source for human genetic variation. Diseases that result from DNA rearrangements have been referred to as genomic disorders. Structural variation of our genome can be responsible for inherited as well as sporadic traits. Rearrangements associated with genomic disorders can be recurrent, with breakpoint clusters resulting in a common sized deletion/duplication, or nonrecurrent and of different sizes. The analyses of breakpoints in the proximal short arm of chromosome 17 (17p) reveal nonallelic homologous recombination (NAHR) as a major mechanism for recurrent rearrangements, whereas nonhomologous end-joining (NHEJ) can be responsible for many of the non-recurrent rearrangements.

Large, population based biobanking initiatives are in full swing in many countries throughout the world. These projects are principally motivated by a desire to understand the myriad factors that have an influence on human health, including the role and interaction of genetics and the environment. But despite this current research activity, a wide range of policy issues remain unresolved. In fact, the continued existence of these issues is quite remarkable - especially when one considers that many have been debated for over a decade and that the actual practice of biobanking, and the implementation of policy frameworks, has continued notwithstanding this lack of consensus regarding key research ethics principles. This talk will focus on two of the most persistent and perplexing of the policy issues associated with biobanks: getting consent and allowing participants to withdraw consent. These are not the only issues associated with biobanks, far from it. But they are the two that have attracted much of the policy attention. In addition, getting and withdrawing consent are fundamental principles in research ethics. Understanding the lack of resolution on these key points seems particularly essential. As such, this talk will primarily focus on the reasons for and ramification of the lack of consensus, including an exploration of whom in the policy community is forwarding the different position. I will analyze factors relevant to the debate, including the role of public perceptions regarding different consent approaches, the law around ownership of samples and health information and the idea that this research is in the public good. In the end, we will see that none of these factors can resolve the debate - at least in the absence of some fundamental and broadly based change in the norms of consent and withdrawal. Despite this reality, I will argue that appropriate governance strategies that specifically engage the issues associated with consent seem the only way forward.

Concurrent Symposia
s02.2 Ethical issues in expanded newborn screening
E. W. Clayton; Vanderbilt University Center for Biomedical Ethics and Society, Nashville, TN, United States.

tion of the current technology since residual transgene expression may alter the biological properties of the resulting iPSCs derivatives or induce malignant transformation. We efficiently derived reprogramming factor-free hiPSCs from several patients with PD using Cre-recombinase excisable viruses and subsequently differentiated these cells into dopaminergic neurons, the cell type most affected in PD. Such factor-free iPSCs maintain a pluripotent state and display a global gene expression profile, more closely related to hESCs than to genetically identical hiPSCs carrying the transgenes. This is consistent with the possibility that residual transgene expression in virus-carrying hiPSCs can affect their molecular and biological characteristics and that factor-free hiPSCs therefore represent a more suitable source of cells for modeling of human disease. (2) Efficient gene targeting strategies to generate markers for differentiation and gene correction. Tracking, accentuating, or accelerating pathological phenotypes in the lab could greatly benefit from cell-typespecific lineage reporters, as well as reliable tools to disrupt, repair, or overexpress genes. However, current techniques of gene targeting are inefficient at best and thus are not routinely used. Here we report the highly efficient targeting of several expressed and silent genes in hESCs and hiPSCs using zinc-finger nuclease (ZFN)-mediated genome editing. s03.2 modeling and treating human genetic disease with induced pluripotent stem (iPs) cells

Ethical Issues Raised by Expanded Newborn Screening Wilson and Jungner argued that newborns should be screened only for serious and well understood disorders that require early intervention of proven efficacy prior to the development of symptoms to avert serious or life-threatening sequelae. In recent years, newborn screening has expanded to include disorders that do not meet these criteria. Many factors have led to this expansion, including the availability of multiplex technologies such as tandem mass spectrometry (MS/MS), parent and provider advocacy, and assertions that the appropriate definition of benefit should be expanded. The technical possibility of performing inexpensive whole genome sequencing of newborns lies in the nottoo-distant future. In this talk, I will consider what limits, if any, ought to be placed on the expansion of newborn screening. I will consider specifically the critiques raised in the United States by the Presidents Council on Bioethics in their report entitled The Changing Moral Focus of Newborn Screening (2008) as well as reports of Swedens experience with newborn screening for alpha-1-antitrypsin in early 1970s. s02.3 Ethical issues in preimplantation genetic diagnosis
G. Pennings; Bioethics Institute Ghent, Ghent University, Ghent, Belgium.

Although prenatal and preimplantation genetic diagnosis are frequently considered as similar, there are two differences that have an enormous impact on the ethical evaluation of the applications: 1) the simultaneous availability of several embryos, and 2) the much larger contribution of the clinician to the parental project. The first difference generates the procreative beneficence principle and the lowering of the indications when in vitro fertilization is indicated for other reasons. Some people label this evolution as a slippery slope because they believe that one adheres or should adhere to the strict medical model. This model is based on full penetrance, extreme severity and invariable expression of the disease. Since there are (almost) no such diseases, it is clear that even the currently accepted practice does not fulfill these conditions. We will illustrate this point by means of two examples: sex selection for diseases with skewed sex ratio and variable sex linked expression and selection of healthy carriers. In the second part, we will consider some of the ethical problems that are generated by the new evolution of genetic screening by means of microarrays. Testing for all chromosomal abnormalities and hundreds of genetic diseases and susceptibilities will confront us with new questions: how to decide which embryo to replace? How to ascertain informed consent before testing? Should information to the parents be limited and if so according to which principles? In essence, the evolution of new techniques always moves in the direction of higher performance but it may confront us with the fact that in reality more information is not necessarily better. s03.1 Human induced pluripotent stem cell based in vitro modeling of Parkinsons disease

A. Raya1,2,3; 1 Institute for Bioengineering of Catalonia (IBEC), Barcelona, Spain, 2ICREA, Barcelona, Spain, 3CIBER-BBN, Barcelona, Spain.

The generation of induced pluripotent stem (iPS) cells by ectopic expression of a defined set of factors has enabled the derivation of patient-specific pluripotent cells and provided valuable experimental platforms to model human disease. Patient-specific iPS cells are also thought to hold great therapeutic potential, although several shortcomings should be addressed before iPS cell technology can be implemented clinically. Here, I will present recent results by our laboratory and others on the usefulness of iPS cells to model human disease, the generation of disease-corrected, patient-specific cells with potential value for cell therapy applications, and novel strategies aimed at the generation of clinically-safe iPS cells. s03.3 Using stem cells to model and treat neurodegenerative diseases
A. D. Ebert; University of Wisconsin-Madison, Department of Neurology, Stem Cell and Regenerative Medicine Center, Madison, WI, United States.

F. Soldner; Whitehead Institute for Biomedical Research, Cambridge, MA, United States.

Human embryonic stem cells (hESCs) as well as induced pluripotent stem cells (iPSCs) derived from somatic cells of patients are predicted to become a powerful tool for biomedical research and may provide a source for cell replacement therapies. Although the realization of hESC/iPSC based therapies is still at an early stage of development, the possibility to model human disease in vitro could make patientspecific hiPSCs immediately valuable. This is particularly relevant for diseases of the central nervous system (CNS) such as Parkinsons disease (PD) which are not always linked to known genetic mutations, where primary neuronal tissue is not available, and in vitro or in vivo animal models only partially recapitulate the underlying pathophysiology. However, there are many technical challenges in generating and manipulating human pluripotent cells before they can be thought to be faithful models of human disease. Here, I will highlight some of the technical challenges and some emerging solutions: (1) Generation of reprogramming factor-free iPSCs to minimize or eliminate genetic alterations in the derived iPSC lines. The use of viruses encoding the reprogramming factors represents a major limita-

Stem cells provide an important tool in which to study human development and disease. Stem cells that naturally carry a genetic mutation, or those that have been genetically manipulated to over-express disease causing mutations, have provided a way to better understand disease processes and mechanisms for a variety of neurological disorders including Down syndrome and Parkinsons disease. The recent advance in stem cell technology in which embryonic stem cell-like cells can be produced by reprogramming somatic cells (termed induced pluripotent stem cells (iPSCs)) has opened yet another window of opportunity to model and study human diseases. iPSCs can now be derived from a multitude of patient populations for both genetically linked and sporadic disorders, including Huntingtons disease, amyotrophic lateral sclerosis, and spinal muscular atrophy. Importantly, these iPSCs can be differentiated into the specific cell types that are affected in these brain and spinal cord diseases. Interestingly, in the case of spinal muscular atrophy, motor neurons derived from patient iPSCs have shown selective vulnerability in the culture dish, suggesting a faithful representation of the human disease process. Not only will iPSCs allow for the examination of mechanisms involved in disease progression, but novel drug compound screening and therapeutic intervention may aid in developing more appropriate treatments for patients with these debilitating diseases.

Concurrent Symposia
s04.1 From Galton to GWAs: the genetic architecture of complex traits
P. Visscher; Queensland Institute of Medical Research, Brisbane, Australia. Hospital, Melbourne, Australia.

The centromere is a highly compacted (and morphologically constricted) structure of the chromosome that is essential for the proper segregation of replicated sister chromatids during cell division. A human centromere typically carries 1-4 Mb of repetitive alpha satellite DNA sequences. Human neocentromeres are fully functional centromeres that are formed ectopically on chromosome arms and are devoid of any alpha satellite DNA. The first case of human neocentromere was described by us on band q25 of a rearranged chromosome 10 in a child with mild speech impediment. To date, over 100 cases involving neocentromere formation have been reported with clinical phenotype ranging from very severe to mild or normal, with some of the cases being directly linked to cancer. In addition to humans, the ability of cells to form neocentromeres has been observed in fly, fungi, and higher plants. Because of their non-repetitive and fully sequenced nature, neocentromeres are highly amenable to molecular analysis. Their study (in parallel with normal centromeres) has led to a better understanding of: (a) the regulatory requirements of the centromere, providing the best evidence that centromere formation is modulated by epigenetic changes at the chromatin level that can occur independently of the underlying DNA sequences. Our work and those of others have also shown that transcription of some of the underlying DNA sequences plays an important role in centromere formation and function; (b) a novel mechanism of cancer development, where studies have shown that a group of atypical lipomas and well-differentiated liposarcomas characteristically carry oncogenic giant supernumerary ring or rod neochromosomes that are mitotically stabilised by the de novo formation of a neocentromere; and (c) a novel mechanism of evolution, where molecular and phylogenetic evidence have shown centromere repositioning via neocentromere formation to be a powerful driving force in chromosome evolution and speciation. s05.2 Neocentromeres in candida albicans

Common complex disease is caused by a combination of multiple genes and environmental effects. Traditionally the genetics of disease has been studied using concepts that refer to the combined effect of all genes (e.g., heritability or sibling risk), for example by studying the recurrence risk or phenotypic correlation of relatives. Genome-wide association studies (GWAS) facilitate the dissection of heritability into individual locus effect. They have been successful in finding many SNPs associated with complex traits and have greatly increased the number of genes where variation is known to affect the trait. However, GWAS have been criticised for not explaining more of the genetic variation that we know exists in the population, and many hypotheses have been put forward to explain the missing heritability. The most plausible explanations are that (i) causal effects are too small to be detected with statistical significance and (ii) causal variants are not well tagged by the SNPs on the commercial arrays, for example because their minor allele frequency (MAF) is lower than genotyped SNPs. Genetic linkage and association analyses are typically implemented as a genome scan, i.e. by generating and testing multiple hypotheses. Such approaches, in particular GWAS based upon SNP markers suffer from a high false negative rate because of the use of stringent false positive thresholds. The use of all GWAS data simultaneously in an estimation rather than hypothesis testing framework is a powerful alternative to hypothesis testing. We show how such whole genome methods can be used to better understand the genetic architecture of complex traits, with applications in height and psychiatric disorders. In particular, we show that using genome-wide marker data can provide unbiased estimates of narrow sense heritability and that GWAS SNP data to estimate additive covariance between unrelated individuals can uncover much more of the genetic variance than methods that rely on hypothesis testing. s04.2 Developments in the genetics of multiple sclerosis progress at last

S. Sawcer; University of Cambridge Neurology unit, Addenbrookes Hospital, Cambridge, United Kingdom.

L. S. Burrack, J. Berman; Department of Genetics, Cell Biology & Development, 6-170 Molecular and Cellular Biology Building, Minneapolis, MN, United States.

Multiple sclerosis is a disabling autoimmune disease of the central nervous system that affects approximately 2.5 million people worldwide (http://www.atlasofms.org/). Little is known about the events that trigger the disease or the factors that govern its highly variable course. Epidemiological studies confirm that genetic factors influence susceptibility but relevant genes have proven difficult to identify. Association with the MHC was established almost 40 years ago but alternate candidate gene studies and whole genome linkage screening were unrevealing. Fortunately the advent of genome wide association studies (GWAS) has revolutionised the genetic analysis of multiple sclerosis. To date 7 GWAS have been completed in the disease and 18 associated variants have been identified. This year the International Multiple Sclerosis Genetics Consortium (IMSGC) and the Wellcome Trust Case Control Consortium (WTCCC) will completed a further, and considerably larger, GWAS involving almost 10,000 patients and 16,000 controls. These new GWAS data will substantially expand the list of associated loci and thereby illuminate pathogenesis. s04.3 Genome-wide association studies in cancer: sorting out the nuggets of truth
S. J. Chanock; Division of Cancer Epidemiology and Genetics, Laboratory of Translational Genomics, Bethesda, MD, United States.

No abstract reiceved as per date of printing, please check the abstract section on the conference website www.eshg.org/eshg2010 for possible updates. s05.1 Neocentromeres in human clinical cases

Centromeres are critical for chromosome segregation and genome stability. Neocentromeres, functional kinetocores that appear at ectopic loci, can form when centromere function is lost at the normal locus. In humans, neocentromeres can arise in cells with gross chromosome rearrangements tohat rescue an acentric chromosome, but have also been described in otherwise healthy individuals where the centromere appears to have been inactivated. The mechanisms of centromere inheritance and neocentromere formation remain unknown. Using the yeast Candida albicans, which has small, regional centromeres, we previously found that disruption of the native centromere in C. albicans results in neocentromere formation (Ketel et al. 2009 PLoS Genetics 5(3):e1000400). These neocentromeres form proximal to the disrupted centromere or at distal loci on the chromosome arms far from the disrupted centromere. Distal neocentromere loci characterized to date share properties of low gene density and flanking repeated DNA sequences. We are also using the C. albicans neocentromere model system to characterize the functional properties of neocentromeres such as the ability to bind cohesin proteins and other chromatin components and the ability to be stably maintained under stressful growth conditions. Finally, in a genome-wide analysis of replication timing, we find that centromeres replicate at the earliest time during S-phase and that DNA that acquires a neocentromere also acquires this early DNA replication property. Thus, C. albicans is a useful model for studying epigenetic activities involving centromeres, such as the establishment of neocentromeres and the maintenance of functional kinetochores at specific DNA loci. s05.3 centromere repositioning in evolution and in humans
M. Rocchi; Dip. di Genetica e Microbiologia, Bari, Italy.

A. Choo1,2,3; 1 Murdoch Childrens Research Institute, Melbourne, Australia, 2Department of Paediatrics, University of Melbourne, Melbourne, Australia, 3Royal Childrens

In recent years we have used large panels of BAC clones to track the evolutionary history of chromosomes in primates and in non-primate mammals. This approach has disclosed an unprecedented phenomenon: the centromere repositioning, that is the movement of the cen-

Concurrent Symposia
tromere along the chromosome without marker order variation. Repositioned centromeres are relatively frequent. In macaque, for instance, 9 out of 21 centromeres are evolutionarily new; in donkey at least 5 such neocentromeres originated after its divergence from zebra (less than 1 million years). A related phenomenon (clinical neocentromeres) has been reported in human clinical cases. Clinical neocentromeres are analphoid centromeres that emerge in ectopic chromosomal regions. Usually they stabilize supernumerary acentric chromosome which have detrimental phenotypic consequences. Studies on the evolution of the chromosomes where clustering of neocentromeres were reported (3q, 13q, and 15q for instance) disclosed distinct, intriguing relationships between human clinical neocentromeres and evolutionary neocentromeres. Additionally, examples are now available of centromere repositioning events in humans, disclosed by chance because they do not result in phenotypic anomalies. s06.1 Dysruption of long-distance highly conserved non coding sequences at the sOX9 locus
S. Benko1, J. Amiel1, A. Munnich1, D. R. Fitzpatrick2, S. Lyonnet1; 1 Dpartement de Gntique et Unit INSERM U-781 , Universit Paris Descartes Hpital Necker, Paris, France, 2MRC Human Genetics Unit, Institute of Genetic and Molecular Medicine, Western General Hospital, Edinburgh, United Kingdom.

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model could be regarded as a novel mutational mechanism causing human congenital malformations, and understanding it will certainly provide a powerful tool in establishing etiology for a broader range of human diseases. s06.2 clustered gene co-regulation and enhancer sharing can be modulated by developmentally regulated chromatin loops
J. Tena1, E. Alonso2, E. de la Calle-Mustienes1, E. Splinter3, W. de Laat3, M. Manzanares2, J. L. Gomez-Skarmeta1; 1 Centro Andaluz de Biologa del Desarrollo, Sevilla, Spain, 2Fundacin Centro Nacional de Investigaciones Cardiovasculares Carlos III,, Madrid, Spain, 3 Hubrecht Institute-KNAW & University Medical Center Utrecht, Utrecht, Netherlands.

One of the key discoveries of vertebrate genome sequencing projects has been the identification of non-coding elements that remained evolutionarily conserved, and thus likely functional. Interestingly, two thirds of them do not correspond to transcribed gene sequences (exons and UTRs); they have been named conserved non-coding sequences (CNCs) and represent a vast amount of DNA (3% of the human genome). Interestingly, enrichment for CNCs has been demonstrated within gene deserts nearest to physically isolated genes known or suspected to be important developmental regulators. It has been suggested that in these cases CNCs may represent regulatory elements (enhancers or suppressors) necessary for the correct spatiotemporal expression of these genes needed for embryonic development, and acting as modular, sometimes combinatorial, tissue-specific enhancers of gene transcription. In that particular context, we will discuss recent findings from our groups regarding: - A common non-coding enhancer genomic variant in a highly conserved sequence located in a non-coding region of the RET gene, altering the binding of a transcription factor expressed in neural crest cell precursors to the enteric nervous system, which would predispose to Hirschsprung disease. - More recently, the discovery of long-distance disruption of enhancer CNCs on both side of the SOX9 gene coding sequences in Pierre Robin sequence (PRS), a common orofacial cleft anomaly with mandibular hypoplasia. The existence of a PRS locus at 17q24 was supported by both linkage analysis and mapping of independent translocation breakpoints that cluster 1.06-1.23 Mb upstream of SOX9. Also, microdeletions or point mutation involved CNCs capable of driving mandibular expression in transgenic mouse embryos. Moreover, the pattern of histone modifications associated with both the centromeric and telomeric regions suggests tissue-specific enhancer function. ChIP experiments demonstrated that a mutated or deleted CNC binds endogenous MSX1 protein. In addition, a human CNC mutation both alters MSX1 binding and abrogates enhancer function in a mandibular mesenchymal cell line. Our data, combined with existing evidence from human and animal phenotypes, strongly suggests that the disruption of distant, tissue-specific regulatory elements, required for the normal development of the mandibula, perturbs embryonic expression of SOX9 and accounts for the PRS phenotype. These observations suggest that the domains to study for genomic alterations, resulting in tissue-specific misregulation of a developmental gene and a subsequent malformation, should be much broader than traditionally investigated. They also results strongly suggest that genomic alteration of highly conserved non-coding elements of the genome, located near to, or at a long distance from, coding sequences of a gene might alter gene expression in a tissue-specific and timing-specific manner. These evolutionarily constrained regions of the genome are under purifying selection for function, and with no protein coding activity, may be disrupted in a modular fashion as many such regulatory elements surround master developmental genes. This

Gene clusters are paradigms to study transcriptional regulation during development. Here, we present a general map of enhancer distribution along the 2 Mb of DNA spanning the IrxA cluster produced by means of transgenic Xenopus, zebrafish and mouse embryos. Using Chromatin Conformation Capture, we demonstrate that enhancer sharing is widespread within the cluster, which explains the common expression domains of IrxA genes in particular tissues and the evolutionary conserved architecture of the cluster. We also identify an insulator and two chromatin loops within the cluster that may help partition it in two independent regulatory domains in certain cell types. We finally show that this topology predicts gene expression in cases where cluster organization has been disrupted during evolution. We conclude that the regulatory constrains imposed by the linear arrangement of clustered genes in the genome can be modulated by developmentally regulated loops that facilitate the formation of gene-specific regulatory landscapes. s06.3 Far reaching consequences - mechanisms and problems of long range control
S. Mundlos; Institute for Medical Genetics, Charit, Universittsmedizin Berlin and Max Planck Institute for Molecular Genetics, Berlin, Germany.

The vast majority of most genomes consists of non-coding sequence with more or less unknown function. This dark side of the genome contains regions of diverse composition including sequences that are highly conserved throughout evolution. Some of these so called conserved non-coding elements (CNEs) have been identified as essential regulators of gene expression. CNEs are particularly abundant in genes deserts surrounding genes that have important functions during development and may be as far as 1 Mb away from the gene they regulate. Gene regulation is achieved through the binding of transcription factors to the element and subsequent loop formation between the CNE and the genes promotor. Mutations that interfer with the cis regulatory capacity of these elements can thus be expected to result in altered gene expression in a certain cell type at a given time point. We have been investigating the consequences of CNE-controled gene regulation and the effect of mutations using cytogenetics and high-resolution array CGH in mouse models and patients. We identified several molecular mechanisms that cause abnormalities in long range control. These include the disconnection of control elements from their target gene by translocations, changes in presumed transcription factor binding sites by point mutations, and altered gene regulation by deletions, and duplications of CNEs. All abnormalities were detected in patients or mice with congenital malformations, i.e. brachydactyly, triphalangeal thumb-polysyndactyly, Laurin-Sandrow syndrome, Cooks syndrome, or syndactyly. We postulate that these conditions are caused by alterations of fine tuning of gene expression which in consequence disturbs dosage-dependent signalling pathways. Due to the fact that this mutation mechanism interfers only with a certain regulatory event and not the entire gene function, the resulting phenotypes are distict from those associated with mutations in the coding region s07.1 the impact of the early social environment on the adult epigenome

M. Szyf; Department of Pharmacology and Therapeutics , McGill University, Montreal, QC, Canada.

No abstract reiceved as per date of printing, please check the abstract section on the conference website www.eshg.org/eshg2010 for possible updates.

Concurrent Symposia
s07.2 identifying parent of origin effects in the human genome
A. Sharp; University of Geneva, Department of Genetic Medicine and Development, Geneva, Switzerland.

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ceptibility to ASDs; for instance, there are much higher concordance rates for ASDs in monozygotic twins (92%) than dizygotic twins (10%), while recent estimates for the sibling recurrence risk is greater than 15. Although ASDs are highly heritable disorders, they exhibit heterogeneous clinical symptoms and genetic architecture which have hindered identification of common genetic susceptibility factors. Although previous linkage studies, candidate gene association studies and cytogenetic studies have implicated several chromosomal regions for the presence of autism susceptibility loci they have failed to consistently identify genes or genomic loci that increase risk of ASD presentations. Using the genome wide association study (GWAS) approach, we have recently identified common genetic variants between two cadherin genes (CDH10 and CDH9) as associated with ASDs (Wang et al, Nature, 2009), as well as a collection of rare copy number variants in neuronal cell-adhesion genes (Glessner et al, Nature, 2009). The discovery cohorts in the GWAS contains 780 families (3,101 subjects) with affected children, and a second cohort of 1,204 affected subjects and 6,491 control subjects, all of whom were genotyped by us in the Childrens Hospital of Philadelphia (CHOP), representing the largest ASDs genetics studies ever performed. The results from these studies and our ongoing search for the causal variants and their potential influence on the neuronal cell-adhesion molecules in the pathogenesis of ASDs will be presented. S08.3 Behavioural genetics in mice

No abstract reiceved as per date of printing, please check the abstract section on the conference website www.eshg.org/eshg2010 for possible updates. s07.3 Using c. elegans to study chromatin regulators involved in human disease
I. J. Latorre1, M. Cheung1, J. Garrigues2, A. Vielle-Canonge1, T. Takasaki2, S. Strome2, J. Ahringer1; 1 Gurdon Institute, The Wellcome Trust/Cancer Research UK, Cambridge, United Kingdom, 2University of California, Santa Cruz, CA, United States.

Regulation of chromatin structure plays a central role in transcriptional control. A large number of chromatin regulating enzymes and complexes are known, however, their mechanisms of action are poorly understood. Global chromatin factor mapping and loss of function studies in single-celled yeast have provided important insights, but there is still little information on genome-wide targets and functions in multicellular organisms. Importantly, animals contain many chromatinregulating complexes not found in yeast, such as the histone deacetylase NuRD chromatin-remodelling complex, and the DRM complex, which includes the tumor suppressor Retinoblastoma. Components of both of these complexes have been implicated in human disease. C. elegans has many features that make it well-suited for studies of chromatin regulation. Of particular note are its small well-annotated genome (30X smaller than human), the ease of RNAi, and the rich resource of chromatin mutants for loss of function studies. Importantly, C. elegans has a complement of chromatin factors very similar to that of humans, allowing investigations of chromatin function in a multicellular organism. As a step toward understanding the functions of chromatin proteins implicated in disease, we are identifying their patterns of binding in C. elegans genome-wide using ChIP-chip and ChIP-seq. In addition, to provide a framework for these studies, we are also generating genome-wide maps of the locations of histones and histone tail modifications. S08.1 Genomic advances in Schizophrenia

J. Flint; The Wellcome Trust Centre for Human Genetics, Oxford, United Kingdom.

No abstract reiceved as per date of printing, please check the abstract section on the conference website www.eshg.org/eshg2010 for possible updates. s09.1 Prenatal diagnosis of skeletal dysplasias

S. Unger; Centre for Pediatrics and Adolescent Medicine, Freiburg University Hospital, Freiburg, Germany.

M. J. Owen; MRC Centre for Neuropsychiatric Genetics and Genomics , Cardiff University, Cardiff, United Kingdom.

Recent studies have supported the hypothesis that the high heritability of schizophrenia reflects a combination of relatively common alleles of small effect and some rare alleles with relatively large effects. Genome-wide association studies have identified several risk loci at genome-wide levels of significance as well as evidence for a substantial burden of common risk loci. Moreover these recent findings suggest genetic overlap with bipolar disorder which has traditionally been assumed to be genetically distinct from schizophrenia. Genome-wide studies of at least one class of relatively uncommon variant, submicroscopic chromosomal abnormalities often referred to as copy number variations (CNVs), suggest that these confer high risk of schizophrenia. There is evidence both for an increased burden of large (>100kb) and rare (MAF <1%) CNVs in schizophrenia and that risk is conferred by a number of specific large CNVs (including deletions at 22q11.2, 1q21.1, 15q13.2 and 15q11.2 and duplications of 16p11.2) as well as by deletions of NRXN1 which encodes the synaptic scaffolding protein neurexin 1. Many of these CNVs have been implicated in autism, mental retardation, epilepsy and other neurodevelopment disorders. The implications of recent findings for the pathogenesis and noslogy of schizophrenia and related disorders will be discussed. S08.2 Genome-Wide Association Studies in Autism Spectrum Disorders

The delineation of skeletal dysplasias originated and still relies heavily on radiographic findings. Translation of this knowledge to ultrasound based diagnosis is not always evident or simple. Ultrasound provides an indirect image that is often focused on details and it can be difficult to get an overview (a form of prenatal babygram). For genetic counseling purposes it is important to distinguish lethal and non-lethal forms of dysplasia. Certain parameters, such as degree of long bone shortening, chest size relative to abdomen, and presence or absence of hydrops or other anomalies, are key to this interpretation. Few signs are specific (and none are pathognomonic) thus a systematic approach to the skeleton is needed. This allows for the best possible chance of diagnosis but this is not always possible prenatally and even the most experienced centers must often give generalized counseling. This talk will review the most common diagnoses as well as some of the pitfalls of ultrasound diagnosis. s09.2 What you see depends on what you look at Genetics Fetal ultrasound perspectives
R. Achiron; University of Tel Aviv, Tel Hashomer, Israel.

H. Hakonarson; Center for Applied Genomics, Childrens Hospital of Philadelphia, Philadelphia, PA, United States.

Autism spectrum disorders (ASDs) represent a large group of childhood neurodevelopmental, neuropsychiatric disorders characterized by restricted and repetitive patterns of interests and behavior, limited verbal communication and impairment of social interaction. Several sources of evidence suggest strong a genetic component in the sus-

Fetal medicine is a new evolving profession which requires a multidisciplinary approach to confront with various fetal diseases. Objective: To review the perspectives of Genetic counselling and Sonographic evaluation in fetuses with abnormalities detected in utero. Methods: A retrospective survey of stimulating and interesting cases will be presneted. Results: Four fetuses representing the following topics are described: 1) New Technology and Pandora Box 2) One Sees what One Knows 3) What you see is the Tip of the Iceberg 4) Not Just Images Conclusion: In modern obstetrics Genetic and fetal medicine collaboration is necessary for enhancing diagnosis and promoting accurate management.

Concurrent Symposia
s09.3 Use of acGH in prenatal diagnosis
I. B. Van den Veyver; Baylor College of Medicine, Obstetrics and Gynecology and Molecular and Human Genetics, Houston, TX, United States.

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s10.2 transitions between research and clinical practice: Families experiences in a gene-hunting study

Current prenatal cytogenetic diagnosis for chromosomal abnormalities is performed by karyotyping of fetal cells from chorionic villi or amniotic fluid to detect any aneuploidy and structural genomic rearrangements of larger than ~5 Mb. In addition, fluorescence in situ hybridization is used for rapid detection of common aneuploidies compatible with live birth or detection of deletion and duplication syndromes for which there is high clinical suspicion because of family history or prenatal ultrasound findings. In contrast, array-based Comparative Genomic Hybridization (aCGH) can survey the entire genome for submicroscopic deletions and duplications, aneuploidy and other unbalanced chromosomal abnormalities. It is now widely used in the genetic evaluation of pediatric patients and is being increasingly applied to prenatal diagnosis where it is already making significant impact. Principles of aCGH and different platforms that can be used for prenatal diagnosis, with their benefits and challenges will be briefly reviewed. Our recent experience of over 600 cases suggests that aCGH-detection of copy number alterations of clinical significance from prenatally obtained samples is as high as 8%, depending on the indication, with more than 2% not otherwise detectable. The challenges of pre- and post-test counseling, cost, and potential for finding copy number changes of unknown significance continue to incite debate about the benefit of widespread use of aCGH for prenatal diagnosis; a large multicenter trial is ongoing in the US to address some of these issues. However, the superior diagnostic power of aCGH, the small number of findings of uncertain significance (1% or less) most of which can be resolved by analyzing parents or by using data from ever-growing experience with diagnostic aCGH, and the decreasing cost, will likely spur universal acceptance of aCGH as a first-line prenatal diagnostic test for fetal chromosomal abnormalities in the near future. s10.1 Patients Organizations Engagement in War on Rare Genetic Diseases: Scientific Activism and New Forms of sociality
V. Rabeharisoa; Ecole Nationale Superieure des Mines de Paris, Paris, France.

H. Statham1, M. Ponder1, M. P. M. Richards1, N. Hallowell2, L. Raymond3; 1 Centre for Family Research, University of Cambridge, Cambridge, United Kingdom, 2Public Health Sciences, University of Edinburgh, Edinburgh, United Kingdom, 3Dept of Medical Genetics, Cambridge Institute for Medical Research, Cambridge, United Kingdom.

The GOLD study (Genetics of Learning Disability, a joint project undertaken by the Department of Medical Genetics (University of Cambridge) and The Wellcome Trust Sanger Institute) to identify novel gene mutations on the X chromosome, recruited patients and families if: the family had multiple male individuals affected by significant Intellectual Disability the pattern of occurrence of the Intellectual Disability suggested an X-linked inheritance pattern all genes known to be responsible for Intellectual Disability had been tested for and excluded. This paper reports some of the findings of complementary research with a subset of families who were part of the GOLD study. The objectives were to explore and document the experiences, beliefs, understandings, attitudes and behaviours of family members while they were participating and afterwards. Analysis revealed similarities between routine clinical practice and the genetic research in terms of process and desired outcome. Thus, in trying to identify the nature of the Intellectual Disability prior to participation in the GOLD study, the diagnostic process would have been to take blood and for the blood to be investigated for known genes. The process when the GOLD research was undertaken would also have been to take blood and to investigate the DNA extracted from the blood, but for as-yet unknown gene mutations. Those undergoing testing in either clinical practice or through research sought a genetic diagnosis for a variety of reasons, the most common of which was to allow informed reproductive choices for themselves and other family members. These similarities raise important issues for clinicians involved with patients moving between clinical practice and research with regards consent processes at the beginning of research and the information and care given to participants at the end of research when they again become patients. s10.3 Genetics and mental illness: perceptions of affected individuals and their family members
J. Austin; University of British Columbia, Vancouver, BC, Canada.

From the 1980s onwards, both in Europe and North America, an increasing number of patients organizations actively engage in the collection and circulation of knowledge on their conditions. Patients organizations concerned with rare genetic diseases offer a striking illustration thereof. Some of them even intervene in research activities and contribute to the shaping of research agenda on their diseases. This communication aims at documenting this phenomenon and highlighting its impact on the governance of knowledge, as well as on the dynamics of patients organizations movements. I will first show that patients organizations engagement in research constitutes a watershed in the history of patients organizations. They do no longer content to provide social and emotional support to their members and to advocate for their rights. They claim to take part in war on their diseases, alongside specialists. In the area of rare genetic disorders, patients organizations are particularly assertive. Due to the scarcity of knowledge on their conditions, the limited number of specialists interested in, and the difficulties for making rare genetic diseases a public health issue, patients organizations soon decided to engage in research in order to foster war on their conditions. Drawing on various case studies, I will then detail different configurations of patients scientific activism. In the area of rare genetic diseases, patients organizations do not only provide financial support to research. They mobilize patients experience and act as lay-experts in the process of knowledge production. They thus nurture specialists expertise with patients experience from bench to bedside, and give raise to hybrid forms of knowledge on their diseases. Finally, I will show that patients scientific activism does not only transform the very nature of knowledge and research activities. It also impinges on patients identity and lay ground for new forms of sociality that the American anthropologist Paul Rabinow terms biosociality. To open the general discussion, I will offer a few examples and I will reflect on the social, political and ethical stakes pertaining this new articulation between genetics and society.

To date, there has been little in the way of systematic effort to provide education about what is known (based on research) about the causes of mental illnesses for affected individuals and their families. However, because understanding cause of an illness is critical to adapting to it, in the absence of being provided with an explanation affected individuals and families create their own explanations, based on their experience. For mental illness, these causal explanations often invoke powerful negative emotions like guilt and shame, and can contribute to feelings of powerlessness over the illness. This presentation will review the potential psychosocial consequences of providing genetic counseling for individuals with severe psychiatric illness and their family members, illustrated with case examples from an ongoing randomized controlled trial. s11.1 mechanisms of miRNA-mediated gene silencing

E. Izaurralde; Max-Planck-Institute for Developmental Biology, Tuebingen, Germany.

MicroRNAs (miRNAs) are genome-encoded ~22 nucleotide-long RNAs that silence gene expression post-transcriptionally by base pairing with the 3 untranslated regions of target mRNAs. To exert their function, miRNAs associate with Argonaute proteins (AGOs) in miRNAinduced silencing complexes (miRISCs), which silence the expression of mRNAs containing partially or fully complementary miRNA-binding sites. In animals, most miRNAs are only partially complementary to their targets. In this case, our group has shown that the AGO proteins are not sufficient to mediate silencing and require interaction with proteins of the GW182 family. We have also shown that AGO-GW182 complexes mediate silencing by promoting translational repression and mRNA deadenylation catalyzed by CAF1-CCR4-NOT, the major cytoplasmic deadenylase complex. Deadenylation decreases transla-

Concurrent Symposia
tion efficiency and, in somatic cells, commits the mRNA to decapping and 5-to-3 exonucleolytic degradation. Our analysis of GW182 protein function has revealed two domains critical for silencing: an N-terminal GW-repeat-containing region conferring binding to AGOs, and a bipartite silencing domain, consisting of Mid and C-terminal regions, which elicits translational repression and degradation of miRNA targets. Exactly how the bipartite silencing domain of GW182 proteins interferes with translation and accelerates deadenylation is not completely understood. We have recently started to address this question by showing that the silencing domains of GW182 interact with the cytoplasmic poly(A)-binding protein 1 (PABPC1), suggesting GW182 proteins are PABP-interacting proteins (Paips) that interfere with the function of PABPC1 in translation and mRNA stabilization. s11.2 the hidden layer of noncoding RNA in the epigenetic control of human development and cognition s12.1 What can be learned from a large clinical cohort
S. Bergmann; Department of Medical Genetics, University of Lausanne, Lausanne, Switzerland.

J. S. Mattick; Institute for Molecular Bioscience, The University of Queensland, Brisbane, Australia.

Bioinformatic, genomic and experimental evidence all suggest that the genetic programming of humans and other complex organisms has been misunderstood for the past 50 years, because of the assumption - largely true for the unicellular prokaryotes, but not for multicellular eukaryotes - that most genetic information is transacted by proteins. The human genome specifies the development of an anatomically and cognitively complex individual comprised of 100 trillion cells with hundreds of different and precisely sculptured muscles, bones and organs, including the brain, but which contains only about 20,000 protein-coding genes, similar in number and largely orthologous with those in nematodes that have only 1,000 somatic cells. On the other hand, the extent of non-protein-coding DNA increases with increasing complexity, reaching 98.8% in humans, suggesting that much of the information required to program development may reside in these sequences. Moreover it is now evident the majority of the mammalian genome is transcribed, mainly into non-protein-coding RNAs (ncRNAs), and that there are tens if not hundreds of thousands of long and short RNAs in mammals that show specific expression patterns and subcellular locations. Our studies indicate that these RNAs form a massive hidden network of regulatory information that regulates epigenetic processes and directs the precise patterns of gene expression during growth and development. It also appears that RNA is central to brain development, learning and memory, and that animals, especially primates, have developed sophisticated RNA editing systems to modify hardwired genetic information in response to experience, that in turn can modulate epigenetic memory, some of which may be inherited. Thus RNA may represent the computational engine of the cell and the substrate for epigenome-environment interactions. Moreover, what was dismissed as junk because it was not understood may hold the key to understanding human evolution, development and cognition, as well as our individual differences and susceptibilities to complex diseases. s11.3 system genetics of non-coding RNA
L. Steinmetz; EMBL Heidelberg, Heidelberg, Germany.

The Cohorte Lausannoise (CoLaus) is a random population sample of more than 6000 individuals who were genotyped for 500.000 Single Nucleotide Polymorphisms (SNPs) using Affymetrix SNP-microarrays. Besides these genotypic markers also a large number of clinically relevant parameters were measured. Comparing the country of origin of these individual with the projection of their genotypic profile onto the principal components of the entire genotypic dataset revealed an astonishingly close correspondence between genetic and geographic distances. Indeed, a geographical map of Europe arises naturally as an efficient two-dimensional summary of genetic variation in Europeans (see Figure). Whole-genome association studies for height, bodymass-index, serum lipid and calcium concentrations, blood pressure and other clinical phenotypes using classical scans testing one SNP at a time elucidated many loci with highly significant associations, which are promising candidates towards unraveling mechanisms of actions and malfunction. Yet, like in many other studies, together these variants only explain a small fraction of the phenotypic variance, indicating that we still miss a comprehensive picture of: (a) what are the causal variants, (b) what effects are attributed by rare variants and/or copy number variations, (c) what fraction of the variance can be explained by SNP-SNP or SNP-environment interactions, and (d) what are the intrinsic limitations of currently used algorithms in dealing with very large sets of genotypic and phenotypic data, which are partially incomplete or noisy. I will outline our research dealing with these challenges. s12.2 Human genetics from genes to complex networks
R. Xavier; Andrew D. Smith, Department of Biology, Boston, MA, United States.

No abstract reiceved as per date of printing, please check the abstract section on the conference website www.eshg.org/eshg2010 for possible updates.S12.3 Pathway Discovery in Adipocyte Biology Using Epigenomics
E. D. Rosen; Beth Israel Deaconess Medical Center, Boston, MA, United States.

An unanticipatedly large proportion of eukaryotic genomes is transcribed. By profiling genome-wide transcription at high resolution on both strands of the complete yeast genome we have found hundreds of novel intergenic and antisense non-coding RNA transcripts. By comparing transcriptome profiles across multiple conditions and strains with shuffled genotypes, we have defined an annotated set of differentially expressed non-coding transcripts revealing differences in structure and level. Hundreds of non-coding RNAs appear to arise from an inherent bi-directional transcription from eukaryotic promoters. Many others originate from 3 nucleosome depleted regions of genes and are transcribed antisense to the open reading frame. The arrangement and regulatory patterns of these transcripts suggest mechanisms of how they could be regulated and how they could regulate the expression of genes.

The epidemic of obesity and Type 2 diabetes has thrust the biology of the adipocyte into the forefront of biomedical research priorities. We are interested in identifying the transcriptional basis by which adipocytes govern their behavior. To this end we have utilized approaches that leverage maps of epigenetic alterations in adipose tissue to predict novel transcriptional pathways. We have generated genome-wide chromatin state maps, PPAR and CTCF localization maps and gene expression profiles from both murine and human models of adipogenesis. These data provide unprecedented views of chromatin remodeling during cellular differentiation, and allow identification of thousands of putative pre-adipocyte- and adipocyte-specific cis-regulatory elements based on dynamic chromatin signatures. We find that the specific locations of most such elements differ between the two models, including at orthologous loci with similar expression patterns. Based on sequence analysis and reporter assays, we show that these differences are determined in part by evolutionary turnover of transcription factor motifs in the genome sequences, and that this turnover may be facilitated by the presence of multiple distal regulatory elements at adipogenesis-dependent loci. Finally, we also utilize the close relationship between open chromatin marks and transcription factor motifs to identify and validate several novel regulators of adipogenesis and lipid homeostasis. s13.1 can breast cancer mortality be reduced with genetic testing?

P. Hall; Dept. Medical Epidemiology and Biostatistics, Karolinska Institutet, Stockholm, Sweden.

No abstract reiceved as per date of printing, please check the abstract section on the conference website www.eshg.org/eshg2010 for possible updates.

Concurrent Symposia
s13.2 common genetic variants and cancer risks for BRcA1 and BRcA2 mutation carriers

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noma (Australia and Canada) and entero-pancreatic endocrine tumors (Italy); pediatric brain cancers (Germany); prostate cancer (Canada and United Kingdom); and renal cancer (European Union and France). Each project is expected to involve specimens (tumor plus normal) from approximately 500 patients. Over time, additional nations and organizations are expected to join the ICGC. Over the next ten years, the ICGC expects to produce comprehensive catalogues of the full range of genetic mutations involved in 50 types of cancer (i.e. 25,000 cancer and 25,000 germline genomes), with key factors being the ability to detect all mutated cancer genes, data at the level of individual DNA bases, application of common standards for pathology and technology and comparison data from matched, non-tumour tissue. The ICGCs informed consent and ethical oversight policies state that cancer patients enrolled in an ICGC-related study should be informed that their participation is voluntary, that their clinical care will not be affected by their participation and that data obtained from analyses using their samples will be made available to the international research community. ICGC projects use common standards of data collection and analysis. In April 2010, the ICGC launched its data portal at www.icgc.org, with the release of several cancer genome datasets that are freely available to the global research community. In my presentation, I will use the examples of pancreatic cancer genome datasets generated by the Australian and Canadian teams. s14.3 Next Generation Human Genetics

A. C. Antoniou, on behalf of the Consortium of Investigators of Modifiers of BRCA1/2 (CIMBA); Centre for Cancer Genetic Epidemiology, Department of Public Health and Primary Care, University of Cambridge, Cambridge, United Kingdom.

Several lines of evidence suggest that genetic factors modify cancer risks for BRCA1 and BRCA2 mutation carriers. Past studies concentrated on variants in candidate genes thought to be functionally relevant to the diseases. However, these have not been very successful and most studies were too small to provide enough power to detect the modest associations that are likely to be present. Recently, polymorphisms identified through genome-wide association studies of unselected cancer patients and controls have been shown to be associated with cancer risk in large studies by the Consortium of Investigators of Modifiers of BRCA1/2 (CIMBA). This approach identified seven genetic variants (in FGFR2, TOX3, MAP3K1, LSP1,2q35, 5p12 and SLC4A7) that are associated with the risk of breast cancer and one variant (in BNC2) that is associated with ovarian cancer risk for BRCA1 and/or BRCA2 mutation carriers. Differential associations were found between these variants and breast cancer risk for BRCA1 and BRCA2 mutation carriers. These are in line with differences observed in the associations between these polymorphisms and different disease subtypes in the general population and suggest that studies of mutation carriers may be useful for identifying genetic variants associated with different disease subtypes in the general population. All polymorphisms appear to interact multiplicatively on breast cancer risk for mutation carriers. Based on the joint genotype distribution of the 7 risk associated SNPs in BRCA2 mutation carriers, the 5% of BRCA2 mutation carriers at highest risk were predicted to have a probability of 80-96% of developing the disease by age 80, compared with 4250% for the 5% of carriers at lowest risk. Such risk differences may be sufficient to influence the clinical management of mutation carriers and suggest that this is may be one of the first clinically useful impact of common, low penetrance variants identified through genome wide association studies. s13.3 is breast cancer prognosis inherited?

D. Nickerson; University of Washington, Department of Genome Sciences, Seattle, WA, United States.

No abstract reiceved as per date of printing, please check the abstract section on the conference website www.eshg.org/eshg2010 for possible updates. s15.1 Using simple cells to model complex Diseases

S. L. Lindquist; Whitehead Institute for Biomedical Research and HHMI, Dept. of Biology, MIT, Cambridge, MA, United States.

H. Nevanlinna; Department of Obstetrics and Gynecology, Helsinki University Central Hospital, Helsinki, Finland.

No abstract reiceved as per date of printing, please check the abstract section on the conference website www.eshg.org/eshg2010 for possible updates. s14.1 the 1000 Genomes project

R. Durbin; Wellcome Trust Genome Campus, Cambridge, United Kingdom.

No abstract reiceved as per date of printing, please check the abstract section on the conference website www.eshg.org/eshg2010 for possible updates. s14.2 international cancer Genome consortium

T. Hudson; Ontario Institute for Cancer Research, MaRS Centre, Toronto, ON, Canada.

The International Cancer Genome Consortium (ICGC) is coordinating an international-scale research effort to obtain a comprehensive description of genomic, transcriptomic and epigenomic changes in the major forms of cancer. This information will lead to better ways of diagnosing, treating and preventing cancer. The ICGC was launched in April 2008, with the announcement of the proposed strategies and policies to the international scientific community to enable funding agencies and research groups to plan their participation within the ICGC. As of April 2010, the ICGC has received commitments from funding organizations in Asia, Australia, Europe and North America for the following cancer genome projects: acute myeloid leukemia (United States); breast cancer, multiple subtypes (European Union, France, and the United Kingdom); chronic lymphocytic leukemia (Spain); colon cancer (United States); gastric adenocarcinoma (China); glioblastoma multiforme (United States); hepatocellular carcinoma, alcohol and associated etiologies (France) and viral etiologies (Japan); lung cancer (United States); oral cavity cancer (India); ovarian cancer (Australia and United States); pancreatic adenocarci-

It is now clear that an astonishing number of human diseases, especially neurodegenerative diseases, result from basic problems in protein folding. These diseases may appear to have little in common with each other besides their devastating effects on patients and their families. Yet one feature they share is the occurrence of complexes of misfolded, aggregated proteins in affected neurons. For each disease, a different protein is the major constituent of the aggregate: in Parkinsons disease alpha-synuclein, in Alzheimers disease A and tau. We have developed simple cellular models these protein folding disorders by over-expressing human disease-associated proteins in yeast. By combining the unique power of yeast genetics with the highly conserved biology of protein homeostasis in all eukaryotes, we use yeast cells as living test tubes to investigate the mechanisms of toxicity associated with problems in protein folding, trafficking, and degradation and complement these basic studies with transcriptional analysis and high-throughput chemical and genetic screens for toxicity modifiers. Progress will be presented on models for the misfolding of -syn and A. Supported by HHMI and NIH grants NS038372, NS060957. s15.2 C. elegans models for neurodegenerative diseases

E. Nollen; Department of Genetics, University Medical Centre Groningen and University of Groningen, Groningen, Netherlands.

Various age-related neurodegenerative diseases, including Parkinsons disease, polyglutamine expansion diseases and Alzheimers disease, are associated with the accumulation of misfolded proteins in aggregates in the brain. However, how and why these proteins form aggregates and cause disease is still poorly understood. Using the nematode worm Caenorhabditis elegans to model these diseases and high-throughput genetic screens, we have identified genes that modify aggregation of the disease proteins and their toxicity. We have recently identified an evolutionarily highly conserved modifier of aggregation, moag-4, as a positive regulator of aggregate formation in C. elegans models for misfolding diseases. We have shown that moag-4 drives the formation of compact misfolding intermediates that are required for aggregate formation. We have also shown that loss of moag-4 pro-

Concurrent Symposia
motes longevity in parallel to the core IGF/insulin (IIS) longevity pathway, converging at the IIS transcription factors, daf-16 and hsf-1. Thus, moag-4 has a dual function as a regulator of protein aggregation and of lifespan in C. elegans. moag-4 represents an unexplored protein quality control pathway and since there are two close human orthologs with conserved functions, our results will open up new avenues for research on aging-related neurodegenerative diseases. s15.3 the prion-like aspect of Alzheimers disease

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of parenchymal plaques, and hyperphosphoylated tau intracellularly as neurofibrillary tangles. To understand the mechanisms how abnormal protein processing and aggregation leads to cerebral amyloidosis and tangle formation, cellular dysfunction, and dementia, several transgenic mouse models have been generated. These mouse models have been instrumental to study the induction and spread of the AD lesions and a mechanism reminescent of prions has been suggested. The observation that A structural variants can be induced in vivo in these mouse models intensifies the search of the agent that drives corruptive protein templating in AD pathogenesis. AD lesions and neurodegeneration likely occur many years before the clinical signs of the disease. Thus, an understanding of the earliest and initial events in the development of AD is crucial for the development of diagnostics and early mechanism-based intervention.

M. Jucker; Department of Cellular Neurology, Hertie-Institute for Clinical Brain Research, University of Tbingen, Tuebingen, Germany.

Cerebral proteopathy is a unifying term for cerebral neurodegenerative diseases in which aggregated proteins are abnormally deposited in the brain. The hallmark proteopathy is Alzheimers disease (AD) in which fibrillar amyloid- (A) peptide is deposited extracellularly in the form

Educational Sessions Abstracts of EsHG Educational sessions

Es1.1 Genetics of Female Ovarian Dysfunction
B. C. J. M. Fauser; Department of Reproductive Medicine and Gynecology, University Medical Center, Utrecht, The Netherlands

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data mining tool will be given with an emphasis on human data in general and variation data in particular. Es3.1 Family matters: theory and practice in the communication of genetic information
C. Gaff; Departments of Paediatrics & Medicine, University of Melbourne, Melbourne, Australia.

Ovarian function is regulated by a complex endocrine (and paracrine) system involving the central nervous system, pituitary gonadotropins, and ovarian steroids. Numerous factors are involved such as kisspeptin, GnRH, LH, FSH (its receptors and signal transduction pathways) and many enzymes and intermediate steroids (in the cascade of converting cholesterol to estradiol), and finally steroid receptors. Rare, single gene mutations and resulting phenotypes have been described for all steps imaginable. Well known phenotypes include Kallman syndrome, 21 hydroxylase deficiency, FSH receptor defect, McCune Albright. These experiments of nature greatly improved our understanding of physiology. Normal menopause occurs between 40 and 60 yrs of age. The age of menopause between sisters (or between mothers and daughters) is closely linked, suggesting a strong genetic component. Age of menopause is related to preceding fertility and has important implications for subsequent female health. Some candidate genes have been associated with age of menopause and currently additional information is generated through the use of genome wide association studies (GWA) using large numbers of samples. Early menopause seems linked to poor IVF outcomes, as well as low response to ovarian stimulation. Primary ovarian insufficiency (POI) is often associated with a premature exhaustion of the primordial follicle pool. Turner Syndrome is linked with the most severe form of POI. Several regions have been identified on the X-chromosome critical for normal ovarian function, and additional mutations, SNPs and CNVs are currently being described on the X chromosome. Many transgenic animal models have been generated to further explore roles of specific genes in ovarian function. The current focus of genetic research beyond candidate genes in female reproduction is completely geared towards the elucidation of complex and common benign conditions such as polycystic ovary syndrome (PCOS), endometriosis, uterine fibroids. Es1.2 Genetic Regulation of spermatogenesis

Facilitating family communication about genetics is an integral part of genetic counselling practice. Although the research literature predominantly focuses on the communication of test results, families inevitably discuss other aspects of genetic conditions, sometimes deliberately, often as part of their regular social discourse. It is likely that communication relating to inheritance and genetic conditions conforms to the rules and patterns that govern communication generally in families. Insights from the discipline of family communication may therefore assist practitioners work with families. They may also enhance awareness of the practitioners own communication patterns and expectations. After a brief introduction to genetic counselling practice in this field, participants will be introduced to theories from the discipline of family communication - including Communication Privacy Management and Family Communication Patterns. The relevance of these to genetic counselling will be explored, with case studies used as a basis for discussion. Throughout the workshop, there will be an emphasis on reflective practice. Es3.2 Families and Genomics: A Biopsychosocial model for clinical Practice
J. Rolland; Department of Psychiatry & Behavioral Neuroscience, The University of Chicago, Chicago, IL, United States.

S. Repping; Professor Human Reproductive Biology, Center for Reproductive Medicine, Amsterdam, Netherlands.

No abstract reiceved as per date of printing, please check the abstract section on the conference website www.eshg.org/eshg2010 for possible updates. Es2.1 Finding your feet in the Genome Database World
C. Broud; Universit de Montpellier, Montpellier, France.

No abstract reiceved as per date of printing, please check the abstract section on the conference website www.eshg.org/eshg2010 for possible updates. Es2.2 Browsing Genes and Genomes with Ensembl

B. Overduin; PANDA Coordination & Outreach, EMBL - European Bioinformatics Institute, Hinxton, Cambridge, United Kingdom.

The Ensembl project provides a comprehensive source of annotation for the human genome, along with other species of biomedical interest. Ensembl automatically annotates genomic sequence and predicts the position of genes, to provide a comprehensive range of sequence features and genome wide gene and protein sets. Ensembl also integrates manually annotated gene structures from external sources where available. Apart from the gene sets, Ensembl contains extensive comparative, variation and regulatory information. A rich variety of links to external databases (e.g. OMIM, dbSNP and the NHGRI GWAS catalogue) helps to make Ensembl a key starting and reference point for studies in genetics and molecular biology. Ensembl data are accessible through an interactive web site (http://www.ensembl.org), flat files, the data mining tool BioMart, direct database querying and a Perl API. In this presentation an overview of the Ensembl project and the BioMart

Groundbreaking advances in genomics are identifying genetic components in most major health and mental health disorders. This poses unprecedented challenges for families. How are family and couple relationships being affected by the ability to peer into their possible health and mental health futures? How can we help families use the knowledge of genetic risk to become more resilient and live life more fully? Drawing from his recent book, Individuals, Families, and the New Era of Genetics, Dr. Rolland will first present an overview of his Family System Genetic Illness model to address the psychosocial challenges of genomic conditions for patients and their families, and to help organize this complex biopsychosocial landscape for clinical practice and research. This model clusters genomic disorders based on key characteristics that define types of disorders with similar patterns of psychosocial demands over time. For disorders in which genetic testing is available, core nonsymptomatic time phases with salient developmental challenges are described pre- and post-testing, including a longterm adaptation phase. The FSGI model builds on Rollands Family System Illness model, which identifies psychosocial types and phases of chronic disorders after clinical onset. The FSGI model is designed to be flexible and responsive to future discoveries in genomic research. Dr Rolland then addresses core issues and cultural influences in decision-making about genetic testing, communication with partners, children, and other family members, and living with risk information across the life cycle. Other key issues discussed include: privacy vs. right to know of others at risk, belief conflicts, impact on childbearing decisions, multigenerational themes, and behavioral genetics. Guidelines are provided for healthcare and mental health professionals to help families master these complex challenges. Its utility is discussed for research, preventive screening, family assessment, treatment planning, and service delivery in a wide range of healthcare settings. Es4.1 Alport syndrome

C. Antignac; INSERM Division: U 423, Paris, France.

No abstract reiceved as per date of printing, please check the abstract section on the conference website www.eshg.org/eshg2010 for possible updates.

Educational Sessions
Es4.2 Renal cystic disease
N. V. A. M. Knoers; Radboud University Nijmegen Medical Centre, Department of Human Genetics 417, Nijmegen, Netherlands.

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achieved just by looking at the face. In this educational session the different anatomical structures of the face are explained and you will learn to recognize facial dysmorphisms of the eyes , nose, mouth, ears etc. In a second part of the course syndromes with a recognizable face will be presented. The variable expression of facial features will be demonstrated by showing the mild and severe end of the spectrum. In addition changing of the face with time will be shown. Finally the participants can test their diagnostic skills in a quiz. Es7.1 Overgrowth

Cystic kidney diseases are among the most frequent incurable genetic diseases. They are a clinically and genetically extremely heterogeneous group of disorders, encompassing autosomal dominant, autosomal recessive and X-linked traits, which are all characterized by perturbed tubular architecture leading to the formation of cysts. The most prominent examples are Polycystic kidney disease (PKD) and nephronophthisis (NPHP), which affect both adults and children. In this educational workshop, I will discuss the clinical and genetic aspects of the different types of PKD en NPHP. I will review studies that link these disorders to disturbed structure and/or function of renal primary cilia. Primary cilia are microtubule-based organelles, protruding from the apical surface of almost all cells in the mammalian body, with an antenna-like sensory function. In recent years primary cilia have been shown to play important roles in embryonic development and tissue homeostasis. The ciliary connection also explains the frequent association of extrarenal symptoms with cystic renal disorders, such as liver fibrosis/cysts, retinal degeneration, brain abnormalities, laterality defects, and others. Several examples of these syndromic forms of cystic kidney disease will be highlighted. Es5.1 Genetic deafness

P. Lapunzina; Section of Medical Genetics, Hospital Universitario La Paz, Madrid, Spain.

No abstract reiceved as per date of printing, please check the abstract section on the conference website www.eshg.org/eshg2010 for possible updates. Es7.2 Undergrowth

A. Rauch; University of Zurich, Institute of Medical Genetics, Zurich, Switzerland.

No abstract reiceved as per date of printing, please check the abstract section on the conference website www.eshg.org/eshg2010 for possible updates. ES8.1 Cystic Fibrosis

M. Bitner-Glindzicz; Institute of Child Health, Ear Institute, Research Area Molecular, Clinical, London, United Kingdom.

B. Kerem; The Hebrew University of Jerusaem, Dep. Genetics, Jerusalem, Israel.

No abstract reiceved as per date of printing, please check the abstract section on the conference website www.eshg.org/eshg2010 for possible updates. Es5.2 Non-syndromic and syndromic retinitis pigmentosa

H. Bolz1,2; 1 Institute of Human Genetics , University of Cologne, Cologne, Germany, 2 Bioscientia Center for Human Genetics, Ingelheim, Germany.

Retinitis pigmentosa (RP) is characterized by progressive loss of night vision in adolescence and constriction of the visual field that may ultimatively lead to tunnel vision. Loss of central vision may occur in later life. Most patients are legally blind by the age of 40 years. RP results from loss of retinal rod and cone photoreceptor cells. Diminished retinal function is detectable by electroretinogram long before night blindness, usually the initial symptom, is noticed by the patient. The appearance of abnormal pigment deposits (so-called bone spicule pigmentation) along with retina atrophy/thinning is the hallmark of RP. RP is the leading cause of visual loss in individuals younger than 60 years, with a prevalence of about 1 in 4000. RP is a major cause of blindness. Non-syndromic RP can have different modes of inheritance: autosomal dominant (ADRP, 30-40%), autosomal recessive (ARRP, 50-60%) and X-linked (XLRP, 5-15%). Most isolated cases probably represent recessive disease. Non-mendelian inheritance patterns, such as digenic inheritance and maternal (mitochondrial) inheritance, exist but are probably rare. RP genes encode for a variety of proteins, reflecting the functional complexity of the retina and including gene products hitherto considered crucial for life such as splice factors and an enzyme of the Krebs cycle. RP is part of many syndromes. In particular, it can be observed in several so-called ciliopathies, e.g. Usher- (with deafness), Joubert(a developmental disorder with brain malformations) and Bardet-Biedl syndrome. Some of these retinal ciliopathies bridge monogenic and oligogenic inheritance, and variants in certain genes have been shown to act as modifiers of retinal disease. This presentation will give an overview of RP genes, the spectrum of phenotypes and future developments in therapy and research. Es6.1 the Face Behind the syndrome

The gene responsible for the cystic fibrosis (CF) disease defined as cystic fibrosis transmembrane conductance regulator 9CFTR) was cloned 20 years ago. The aim of cloning the CF gene was to uncover new knowledge that will help to prevent, detect, diagnose and treat patients suffering from the CF disease. In this educational lecture I will overview and discuss: 1. The impact of cloning the CFTR gene on CF incidence and prevalence. 2. Our understanding of the effect of genetic modifiers on disease severity and 3. The current status of therapeutic approaches based on CFTR knowledge. In the first part of the lecture I will show that in most countries, but not all, we succeeded in finding the CF causing mutations in the majority of patients and by this generated the required basis for genetic testing. The incidence of new CF live birth has decreased in recent years in several countries while in others no change is found. I will discuss potential factors leading to this difference. In the second part I will discuss the molecular basis for disease variability among different patients and review the effect of genetic modifier on the disease severity. In the third part I will summarize the therapeutic approaches which are being developed based on CFTR knowledge. This will include the current status of gene therapy for CF, activation of non-CFTR chloride channels, and mutation specific therapies. In summary, a trend for preventions is already found in many countries, better tools for CF diagnosis were developed, and therapeutic approaches are being investigated. Overall, 20 years of progress enabled us to develop new hopes for a better future for CF patients around the world. ES8.2 Immotile cilia

H. Omran; Universittsklinikum Mnster, Zentrum fr Kinderheilkunde und Jugendmedizin, Mnster, Germany.

No abstract reiceved as per date of printing, please check the abstract section on the conference website www.eshg.org/eshg2010 for possible updates.

G. Gillessen-Kaesbach; Institut fr Humangenetik, Universitt zu Lbeck, Lbeck, Germany.

Syndromes are typically diagnosed by a combination of clinical features. In many conditions the facial gestalt is of high diagnostic value. Recognition of syndromes like Cornelia de Lange syndrome, Kabuki syndrome, Treacher Collins syndrome and many others can be

Concurrent Sessions Abstracts of EsHG concurrent sessions

c01.1 tracing the derivation of embryonic stem cells from the inner cell mass by single cell RNA-seq analysis
K. Q. Lao1, F. Tang2, C. Barbacioru1, S. Bao2, C. Lee2, E. Nordman1, X. Wang1, M. A. Surani2; 1 Molecular Cell Biology Division, Life Technologies, Foster City, CA, United States, 2Wellcome Trust/Cancer Research UK Gurdon Institute of Cancer and Developmental Biology, University of Cambridge, Cambridge, United Kingdom.

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Barcelona, Spain, 3CEGEN _ Barcelona Node, Genoma Espaa, Barcelona, Spain, 4Servei Anatomia Patologica, IDIBELL-Hospital de, Barcelona, Spain, 5 Pompeu Fabra University, Experimental and Health Sciences department, Barcelona, Spain.

The molecular mechanism underlying the transition from the inner cell mass (ICM) of blastocysts to pluripotent embryonic stem cells (ESC) is not fully understood. This is partly because of the apparent heterogeneity amongst a small group of cells, which poses difficulties in investigating this question. Using single cell analysis, including RNA-Seq transcriptome analysis at the resolution of single cells, we have analysed the dynamic molecular network within individual cells from the ICM outgrowth and the established ESC. This study has identified molecular changes that accompany this transition. Our study shows that key genes that confer the property of self-renewal are up regulated as ICM cells progress to ESC. We also detected very significant global changes of transcript variants from individual genes, amongst which the general merabolism genes are strongly over-represented. Furthermore, there was a global increase in the expression of repressive epigenetic regulators with a concomitant decrease in gene activators. The unique ESC epigenotype may thus be sustained while retaining an inherent plasticity for differentiation. Moreover, changes in microRNAs result in one set that targets early differentiation genes, and the second set targets ESC specific pluripotency genes to maintain a delicate balance between pluripotency and a capacity for rapid differentiation. A similar paradigm may also subvert normal developmental sequence in specific adult cells during the formation of diseased tissues, including cancers. c01.2* Variation in transcription factor binding among humans
M. Kasowski1, F. Grubert2, C. Heffelfinger1, M. Hariharan2, A. Asabere1, S. Waszak3, L. Habegger1, J. Rozowsky1, M. Shi1, A. E. Urban1, M. Hong1, K. J. Karczewski2, W. Huber3, S. M. Weissman1, M. B. Gerstein1, J. O. Korbel3, M. Snyder1; 1 Yale University, New Haven, CT, United States, 2Stanford University, Palo Alto, CA, United States, 3European Molecular Biology Laboratory, Heidelberg, Germany.

DNA methylation is one of the most remarkable events within the epigenetic mechanisms of gene regulation, development and genetic imprinting in vertebrates. Alterations in the methylation pattern of regulatory regions have been linked to several pathologies, such as cancer and neuropsychiatric disorders (including schizophrenia and Rett syndrome). In addition, there is an emergent interest to elucidate the relationship between the ageing process, neurodegenerative diseases and aberrant patterns of methylation. For this purpose, we have compared the DNA methylation profile along seven relevant brain areas between Alzheimers and Parkinsons disease samples and controls. DNA extracted from prefrontal cortex, amygdala, hippocampus, hypothalamus, pons, substantia nigra and cerebellar vermis, from cases and controls, was bisulphate treated and hybridized on an Illumina Infinium methylation array (HumanMethylation27), covering 27,578 CpG sites located in the regulatory region of 14,475 genes and 110 miRNAs promoters. First, an unsupervised hierarchical analysis was used to detect a methylation pattern dependent on brain area, as previously suggested by Ladd-Acosta et al. (2007). Our data show a clustering of all the cerebellar samples (independent of disease status), confirming the observation made by Ladd-Acosta, although the remaining brain areas did not show such a clear clustering effect. Secondly, we proposed to identify disease and tissue specific methylation changes. We observed methylation differences in genes previously described as related to neurodegenerative disease (APOE, PSEN1, SIRT3, and MAOA/B) or to epigenetic mechanisms, as DNMT1, as well as in novel candidate genes. c01.4 Dissecting the regulatory network of p63 in p-related developmental disorders
J. Zhou, H. van Bokhoven; Radboud University Nijmegen Medical Centre, Nijmegen, Netherlands.

Differences in gene expression may play a major role in speciation and phenotypic diversity. Although variations in gene expression among individuals have been documented, the origins of these differences are not clear, and studies that directly measure differences in transcription factor binding sites among humans have not been performed. We have examined genome-wide variation in transcription factor binding in different individuals and a chimpanzee using chromatin immunoprecipitation followed by massively-parallel sequencing (ChIP-Seq). The binding sites of RNA Polymerase II (Pol II) as well as a key regulator of immune responses, NFB, have been mapped in ten lymphoblastoid cell lines derived from individuals of African, European, and Asian ancestry, including a parent-offspring trio. Using a stringent threshold, approximately 7.5% and 25% of the respective NFB and Pol II binding regions exhibit differences between any two individuals. To understand the underlying basis of the variations, we examined the effect of SNPs and genomic structural variations (SVs) on binding differences among individuals. We find that many binding differences are associated with SNPs and SVs. Comparison of the binding data with gene expression data generated by RNA-Seq revealed that differences in binding often correlate with gene expression differences. Furthermore, comparison of the Pol II human sites with binding sites identified in the chimpanzee suggests a high level of divergence in binding relative to our closest evolutionary neighbor. Our results indicate that many differences in individuals occur at the level of TF binding and provide insight into the genetic events responsible for these differences. c01.3* Genomewide DNA methylation analysis in neurodegenerative disorders

The transcription factor p63 is a key factor in ectodermal development. Heterozygous mutations in p63 give rise to seven clinical conditions with autosomal dominant inheritance in human. These conditions are characterized by different combinations of split hand-split foot malformation, cleft lip/palate and ectodermal dysplasia (ED). Numerous p63 target genes have been reported; however, their contributions to the phenotypes in the patients carrying a p63 mutation have remained unclear. To understand the regulatory network of p63 relevant to patient phenotypes, we used a disease model, human primary keratinocytes established from patients with p63 mutations. A combination of genome-wide expression profiling and DNA-binding analysis by Chromatin immunoprecipation followed by deep sequencing (ChIP-seq) in primary keratinocytes revealed a subgroup of direct novel target genes relevant to ED syndromes. A number of validated p63 binding sites appear to be directly involved in related genetic disorders (cleft lip/palate and split hand-split foot malformation). These binding sites can be found in promoter regions and introns, but in several instances also at distances up to 500 kb from the predicted target gene. Gene expression controlled by p63 binding sites identified in our study was confirmed functionally in transgene reporter assays in zebrafish and mouse. Our study not only increases the repertoire of p63 target genes but also provides a concrete molecular basis to elucidate the disease mechanism of p63 and p63-related developmental disorders. C01.5 Next generation sequencing-based mRNA profiling of total blood in a large human cohort
P. A. C. t Hoen1, J. T. den Dunnen1, E. J. C. de Geus2, D. I. Boomsma2, J. J. Hottenga2, B. W. J. H. Penninx3, G. J. B. van Ommen1; 1 Center for Human and Clinical Genetics - Leiden Genome Technology Center, Leiden University Medical Center, Leiden, Netherlands, 2Department of Biological psychology - Netherlands Twin Registry, VU University, Amsterdam, Netherlands, 3Department of Psychiatry - Netherlands Study of Depression and Anxiety, VU University Medical Center, Amsterdam, Netherlands.

S. Iraola1, R. Rabionet1, G. Roma2, M. Montfort3, S. Carbonell3, I. Ferrer4, X. Estivill1,5; 1 Center for Genomic Regulation (CRG) and CIBERESP, Barcelona, Spain, 2 Center of Genomic Regulation (CRG), Bioinformatics Core Facilities,

With rapidly decreasing sequencing cost, sequence-based gene expression profiling becomes an attractive alternative over array-based studies. We report on one of the first sequence-based studies into the inter-individual variability of gene expression levels in total blood (n=104). Despite the high abundance of reticulocyte-derived hemoglo-

Concurrent Sessions
bin mRNAs (20-80% of reads), the sequencing depth of 102.5 million reads per sample allows for the reliable quantification of mRNAs derived from ~12,000 genes with an expression level down to 0.3 copies per cell. The amount of hemoglobin transcripts shows a significant inverse correlation with white blood cell counts at the time of sample collection. The absolute nature of the expression levels obtained with next generation sequencing, the high sensitivity of the technology, and the presence of cell type-specific transcripts allow for the accurate estimation of the relative amounts of white blood cells, including those for low abundant basophils and eosinophils. Since differences in blood cell content are a major confounding factor in blood-based expression profiling studies, it is essential to correct for these differences before analyzing expression differences between subjects. Unlike array-based studies, sequence-based studies enable the quantification of allele-specific expression using variants in the mRNA-derived sequence reads. We found that the majority of genes demonstrate preferred expression of one of the two alleles. Furthermore, we observed remarkable inter-individual differences in the preference of one allele over the other. Another factor contributing to the inter-individual differences in gene expression is the preferred expression of specific splicing isoforms and/or use of shorter or longer 3-UTRs. c01.6 mRNA-seq transcriptome analysis of human trisomy 21 using monozygotic twins
A. Letourneau1, S. B. Montgomery1, C. Borel1, E. Migliavacca1,2, D. Robyr1, L. Farinelli3, S. Deutsch1, S. Dahoun-Hadorn1, E. T. Dermitzakis1, S. E. Antonarakis1; 1 Department of Genetic Medicine and Development, University of Geneva Medical School and University Hospitals of Geneva, Geneva, Switzerland, 2 Swiss Institute of Bioinformatics, Switzerland, 3FASTERIS SA, Plan-lesOuates, Switzerland.

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stricted to band 16q24.3 have not been reported before Methods: In this study we aimed to characterize the clinical and molecular features of four patients with de novo submicroscopic interstitial 16q24.3 microdeletions ascertained by genome-wide array analysis and to determine the shortest region of overlap (SRO) to identify candidate genes responsible for their overlapping phenotype Results: Clinical features observed in these patients include facial dysmorphisms comprising prominent forehead, large ears, smooth philtrum, pointed chin and wide mouth, variable cognitive impairment, autism spectrum disorder, structural anomalies of the brain and seizures. The common region of overlap of the deletions is only 90 kb and comprises two known genes, Ankyrin Repeat Domain 11 (ANKRD11) and Zinc Finger 778 (ZNF778), and is located approximately 10kb distally to Cadherin 15 (CDH15). This region is not found as a copy number variation in controls. Discussion: We propose that these patients represent a novel and distinctive microdeletion syndrome, characterized by autism spectrum disorder, variable cognitive impairment, facial dysmorphisms and brain anomalies. We suggest that haploinsufficiency of ANKRD11 and/or ZNF778 contribute to this phenotype and speculate that further investigation of non-deletion patients who have features suggestive of this 16q24.3 microdeletion syndrome might uncover other mutations in one or both of these genes. c02.2 Prader-Willi like phenotype in 2pter deletion: a possible imprinted locus.

Trisomy 21 (T21) is the most widely studied model phenotype of whole chromosome aneuploidy. It is likely that the majority of the T21 phenotypes are related to alterations of gene expression. Transcriptome sequencing now provides the opportunity to investigate in details the perturbations of gene expression in T21 cells and tissues. In this study we used fibroblasts derived from a pair of monozygotic twins discordant for T21. For the first time, the use of these samples eliminates the bias of genome variability and thus all transcriptome differences observed are likely to be related to the supernumerary chromosome 21. The transcriptome (polyA+ mRNA) was studied by RNA-Seq; 29 million 76 bp paired-end reads were generated from each sample. We were able to compare the expression of 285550 exons and 28178 genes between the samples. We observed that about 9% of exons are differentially expressed between the two samples (FDR<0.01). As expected, we found that the majority (93%) of chromosome 21 exons are overexpressed in the trisomic twin. Differentially expressed exons and genes, chimeric transcripts, splicing variants and allelic expression imbalances will be presented. We will also compare the data with previous microarray studies in order to investigate which transcriptome differences can be validated by sequencing and truly related to the trisomy 21 per se. C02.1* Identification of ANKRD11 and ZNF77 as candidate genes for autism and variable cognitive impairment in the novel 16q24.3 microdeletion syndrome

M. Doco-Fenzy1, E. Landais1, M. Vincent2, A. Schneider2,3, J. Puechberty2,3, M. Girard3, M. Tournaire3, E. Sanchez4, M. Goossens5, D. Gaillard1, L. Taine6, G. Lefort2,3, P. Sarda2, B. Leheup7, D. Genevive2,4; 1 Service de Gntique, Centre de Rfrence Maladies Rares Anomalies du Dveloppement et Syndromes Malformatifs Est, Hpital Maison-Blanche. CHRU Reims, UFR de Mdecine, Reims, France, 2Dpartement de Gntique Mdicale et chromosomique, Centre de Rfrence Maladies Rares Anomalies du Dveloppement et Syndromes Malformatifs Sud-Languedoc Roussillon, CHRU Montpellier, Universit Montpellier 1, Facult de Mdecine MontpellierNmes, Montpellier, France, 3Plateforme puce ADN, Service de Gntique Mdicale et chromosomique, CHRU Montpellier, Universit Montpellier 1, Montpellier, France, 4Unit Inserm U844, Institut des neurosciences de Montpellier, Montpellier, France, 5Laboratoire de Biochimie Gntique, AP-HP, et INSERM U-841, CHU Henri Mondor, Creteil, France, 6Service de Gntique Mdicale, centre promoteur du rseau tlomres ACLF, CHU Pellegrin, Bordeaux, France, 7Service de mdecine infantile 3 et de gntique clinique, Hpital denfants, CHU de Nancy - Brabois, Nancy, France.

M. H. Willemsen1, B. A. Fernandez2, C. A. Bacino3, E. Gerkes4, A. P. M. de Brouwer1, R. Pfundt1, B. Sikkema-Raddatz4, S. W. Scherer5, C. R. Marshall6, L. Potocki3, H. van Bokhoven1, T. Kleefstra1; 1 Radboud University Nijmegen Medical Centre, Nijmegen, Netherlands, 2 Memorial University of Newfoundland and Provincial Medical Genetic Program, Eastern Health, St. Johns, NL, Canada, 3Baylor College of Medicine, Texas Childrens Hospital, Houston, TX, United States, 4University Medical Centre Groningen, Groningen, Netherlands, 5The Centre forApplied Genomics, The Hospital for Sick Children and University of Toronto, Toronto, ON, Canada, 6The centre for Applied Genomics, The Hospital for Sick Children and University of Toronto, Toronto, ON, Canada.

Introduction: The clinical utilization of array comparative genomic hybridization in the evaluation of patients with intellectual disablity has recently led to the discovery of a number of novel microdeletion and microduplication syndromes. Aberrations of chromosome 16q with clinical relevance have rarely been reported. Interstitial deletions re-

Pure subtelomeric deletion of the short arm of the chromosome 2 is an extremely rare chromosomal anomaly. To date only, 4 patients with a pure 2pter deletion have been reported in the literature. The phenotype of these patients correspond either to a Prader-Willi-like phenotype (severe precocious obesity associated to mental retardation and abnormal behavior in 1 patient) or an Angelman-like phenotype (IUGR, mental retardation, speech delay, microcephaly and seizures in 2 patients). The last patient presented with a MCA/MR phenotype but the 2pter deletion was different and distant from the 3 other patients. In addition, 2 other patients with Prader-Willi-like phenotype and unbalanced translocation leading to a 2pter deletion have been reported in the literature. Here we report on two novel patients with small pure 2pter deletion and Prader-Willi like phenotype. Cytogenetic studies including FISH using BAC-PAC telomeric probes, BAC-PAC Array-CGH and ArraySNP showed small 2pter deletions estimated to 3.15Mb and 1.96 Mb respectively. Both deletions encompassed the TMEM18 gene. Interestingly, a meta analysis of GWAS for obesity found a significative association with a SNP (rs6548238) near to the TMEM18 gene. In addition, Dong et al suggested that 2pter is a possible human obesityrelated imprinted locus. These data suggest that the short arm of the chromosome 2 is an imprinted locus associated with phenotypes similar to those observed at the 15q11.2 imprinted locus. Parental origin of the deletion in our patients is in progress in an attempt to accumulate data in favour of an imprinted locus at 2p25.3.

Concurrent Sessions
c02.3 2q11.2 is a highly penetrant susceptibility locus for neurocognitive deficit

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with obesity or other complex traits. In addition, we hypothesize that clinical symptoms in carriers of the deletion and the duplication represent opposite manifestations of the same pathophysiologic process. In search of these opposite manifestations mediated by gene dosage, we are currently characterizing BMI, eating behavior, cognitive and psychiatric phenotypes in carriers of both types of rearrangements. Preliminary data reveals that carriers of the duplication show a trend towards being underweight which may confirm this hypothesis. c02.5 mesomelia-synostoses syndrome results from deletion of SULF1 and SLCOA1 genes at 8q13

H. Kilpinen1,2, A. Pittman3, M. Storer1,4, J. Dickerson5, B. Garg6, L. Willatt7, J. Rosenfeld8, N. Huang1, T. Fitzgerald1, D. Felik9, C. Ogilvie10, M. Irving10, Y. Shen11, B. Wu11, R. Pfundt12, B. de Vries12, L. Peltonen1,2, M. Hurles1, J. Barrett1, L. Shaffer8, C. Shaw-Smith1,4; 1 Wellcome Trust Sanger Institute, Hinxton, Cambridge, United Kingdom, 2 Institute for Molecular Medicine Finland (FIMM), Helsinki, Finland, 3Institute of Cancer Research, Sutton, Surrey, United Kingdom, 4Institute of Child Health, London, United Kingdom, 5Riley Hospital for Children, Indianapolis, IN, United States, 6Indiana University School of Medicine, Indianapolis, IN, United States, 7 Addenbrookes Hospital, Cambridge, United Kingdom, 8Signature Genomics, Spokane, WA, United States, 9Bay Regional Medical Centre, MI, United States, 10 Guys and St Thomas NHS Foundation Trust, London, United Kingdom, 11 Childrens Hospital Boston, Boston, MA, United States, 12Radboud University Nijmegen Medical Centre, Nijmegen, Netherlands.

One possible explanation for the missing heritability in common disorders such as autism and schizophrenia is the existence of rare alleles of high penetrance, which escape detection in genome-wide association studies. Recent large-scale studies of autism and schizophrenia have implicated copy number variation in the pathogenesis of these disorders 1,2. Previously, a 1.3 Mb microdeletion flanked by segmental duplications at 2q11.2 was reported in a single patient referred for CNV profiling, but no phenotypic information was provided 3. We now report five patients with microdeletions of similar size at this locus (chr2: ~96.0 to ~97.3 Mb, Hg18). All five patients presented with some form of neurocognitive deficit: four with developmental delay, three with autism or autism-like features, and two with epilepsy. We found one further example of this microdeletion in a schizophrenia case in a publicly available CNV dataset 2. No examples of this deletion were identified in ~7,500 controls. Notwithstanding that two of the deletions were inherited from a neurocognitively normal father, (one de novo, testing of parental samples in the remaining two in progress), this appears to be a very rare but highly penetrant susceptibility locus for neurocognitive deficit. Six highly homologous (>97%) segmental duplications (SDs), size 2.5-40 Kb, cluster at the breakpoints. The rarity of this microdeletion syndrome may be a consequence of the relatively short length of these SDs. 1. Marshall CR et al. Am J Hum Genet. 2008 Feb;82(2):477-88. 2. International Schizophrenia Consortium. Nature. 2008 455(7210):237-41. 3. Rudd MK et al.Hum Mol Genet. 2009 18(16):2957-62. c02.4 the multiple phenotypes of the recurrent 593 kb, 16p11.2 rearrangements: regulation of adiposity, language impairment and psychiatric symptoms.
S. Jacquemont1, R. G. Walters2,3, S. Bouquillon4, F. Zufferey1, A. Valsesia5, D. Martinet1, L. Hippolyte1, J. Andrieux4, B. Delobel6, A. I. F. Blakemore2, P. Froguel2,7, J. S. Beckmann1,8; 1 Service de Gntique Mdicale, CHUV, Lausanne, Switzerland, 2Department of Genomics of Common Disease, Imperial College London, London, United Kingdom, 3Department of Epidemiology and Public Health, Imperial College London, London, United Kingdom, 4Laboratoire de Gntique Mdicale, CHRU Lille, Lille, France, 5Departement de Gntique Mdicale, Universit de Lausanne, Lausanne, Switzerland, 6Centre de gntique Chromosomique, Hopital Saint Vincent de Paul, GHICL, Lille, France, 7CNRS, 8090 - Institut de Biologie, Institut Pasteur, Lille, France, 8Dpartement de Gntique Mdicale, Universit de Lausanne, Lausanne, Switzerland.

C. Le Caignec1,2, B. Isidor1, O. Pichon1, R. Redon2, D. Day-Salvatore3, A. Hamel1, L. Kjelln4, C. Kraus5, J. Leroy6, G. Mortier7, A. Rauch8, A. Verloes9, A. David1; 1 CHU Nantes, Nantes, France, 2Inserm, UMR915, Nantes, France, 3Institute for Genetic Medicine, Saint Peters University Hospital, New Brunswick, NJ, United States, 4Department of Medical Biochemistry and Microbiology, Uppsala, Sweden, 5Institute of Human Genetics, University of Erlangen-Nuremberg, Erlangen, Germany, 6Department of Medical Genetics, Ghent University Hospital, Ghent, Belgium, 7Department of Medical Genetics, Antwerp University Hospital, Antwerp, Belgium, 8Institute of Medical Genetics, University of Zurich, Schwerzenbach-Zurich, Switzerland, 9Department of Clinical Genetics and INSERM U676, Robert Debr University Hospital, Paris, France.

Mesomelia-synostoses syndrome (MSS) or mesomelic dysplasia with acral dysostoses Verloes-David-Pfeiffer type is a rare autosomal dominant disorder characterized by mesomelic limb shortening, acral synostoses and multiple congenital malformations. So far, five patients in four unrelated families have been reported worldwide with MMS. Using whole genome oligonucleotide array CGH, we have identified an interstitial deletion at 8q13 in all patients. The deletions vary from 582 kb to 738 kb in size, but invariably encompass only two genes: SULF1, encoding the heparan sulfate 6-O-endosulfatase 1 and SLCO5A1, encoding the solute carrier organic anion transporter family member 5A1. SULF1 acts as a regulator of numerous growth factors in skeletal embryonic development while the function of SLCO5A1 is yet unknown. Breakpoint sequence analyses performed in two families showed non-recurrent deletions. Our results strongly suggest that haploinsufficiency of SULF1 contributes to this mesomelic chondrodysplasia, highlighting the critical role of endosulfatase in human skeletal development. As co-deletion of SULF1 and SLCO5A1 - which does not result from a low-copy repeats (LCRs)-mediated recombination event - was found in all patients, we suggest that haploinsufficiency of SULF1 combined with haploinsufficiency of SLCO5A1 (or the altered expression of a neighbouring gene through a position effect) could be necessary in the pathogenesis of MSS. c02.6 SHOX duplications are associated with type i mayerRokitansky-Kuster-Hauser (mRKH) syndrome

M. Miozzo1, C. Gervasini1, F. Grati2, F. Lalatta3, S. De Toffol2, B. Gentilin3, P. Colapietro1, M. Silvia1, G. Frontino4, L. Fedele4, B. Dallapiccola5, L. Larizza1; 1 Genetica Medica, Dipartimento di Medicina, Chirurgia e Odontoiatria, Universit degli Studi di Milano, Milano, Italy, 2Cytogenetics and Molecular Biology Unit, Laboratorio TOMA, Busto Arsizio, Varese, Italy, 3UO Genetica Medica, Fondazione IRCCS Ca Granda Ospedale Maggiore Policlinico, Milano, Italy, 4UO Ostetrico-Ginecologica I, Universit degli Studi, Milano, Fondazione IRCCS Ca Granda Ospedale Maggiore Policlinico, Milano, Italy, 5Ospedale Pediatrico Bambino Ges, Roma, Italy.

The 16p11.2 deletion has been associated with childhood-onset developmental disorders, macrocephaly and autism in multiple cohorts, while the reciprocal duplication has been associated with microcephaly, schizophrenia and bipolar disorder. We report an association between this deletion and obesity, regardless of the presence of cognitive or behavioral symptoms. This highly penetrant form of adolescent or adultonset obesity was initially observed in 31 carriers of the 593kb deletion ascertained for cognitive deficits. Nineteen similar deletions were identified from GWAS data of 16053 individuals from 8 European cohorts. Such deletions were absent from healthy non-obese controls and accounted for 0.7% of morbid obesity cases (body mass index, BMI 40 kg.m-2 or BMI standard deviation score 4; p = 6.4x10-8, OR = 43.0), demonstrating the potential etiological importance of rare variants with strong effects in common disease. These rare variants, which escape detection by GWAS, might account for a substantial fraction of patients

Purpose: MRKH syndrome is defined as congenital aplasia of the structures derived from the Mllerian ducts in females with a normal 46,XX karyotype and secondary sexual characteristics. MRKH is frequently sporadic, although familial cases with an unknown pattern of inheritance have been described. Isolated (type I) and complex (type II) forms exist. The genetic basis of MRKH is largely unknown. Genetic lesions, including WNT4 point mutations and genomic imbalances, have been identified in a small number of cases. The aim of the study was to identify possible recurrent sub-microscopic imbalances in a cohort of familial and sporadic MRKH cases. Methods: Multiplex ligationdependent probe amplification (MLPA) was used to screen the subtelomeric sequences of all chromosomes in 30 MRKH patients (sporadic n= 27; familial n= 3). Segregation analysis and pyrosequencing were applied to confirm MLPA data from the informative family. Twelve patients with clinical signs of hyperandrogenism were also screened for WNT4 mutations.

Concurrent Sessions
Results: A partial duplication in the Xpter PAR1 region containing the SHOX gene was found in five MRKH patients (familial n=3; sporadic n=2). The duplications were not overlapping and SHOX was never entirely duplicated. Haplotyping in the informative family revealed that the SHOX duplication had been inherited from the normal father and was absent in the two healthy sisters. No WNT4 mutations were identified in the 12 patients with clinical signs of hyperandrogenism. Conclusions: SHOX, which is known to be responsible for Leri-Weill Dyschondrosteosis and Langer Mesomelic Dysplasia, is associated with both familial and sporadic type I MRKH. c03.1 in-depth metabolic characterization of genetic loci underlying serum-lipids

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identified as associated with obesity in patients ascertained for developmental delay - account for ~0.7% of morbid obesity cases in the general population (Walters et al (2010) Nature 463:671-675). We are now investigating other GSV regions, identified from cohorts of patients with extreme obesity phenotypes or with severe early-onset obesity, by carrying out algorithmic analysis of genotyping data from multiple cohorts. For one region, similar GSVs are found in non-obese subjects, calling into question the putative association with obesity; for 4 others, GSVs are not found in our cohorts but we have instead identified haplotype signatures that are associated with (or protective against) obesity; and numerous smaller GSVs have helped to define the limits of putative obesity-associated loci. Of particular note is the identification of further instances of 220kb deletions surrounding SH2B1 (Bochukova et al (2010) Nature 463:666-670), exclusively in obese subjects and accounting overall for ~0.5% of severe childhood obesity. Targeted exome sequencing is being undertaken to screen for additional rare causal variants in these regions. c03.3* multiple common genetic variants for coeliac disease influencing immune gene expression

A. K. Petersen1, S. Y. Shin2, W. Rmisch-Margl3, G. Zhai4, K. Small4, R. WangSattler1, E. Grundberg2,4, J. S. Ried1, A. Peters1, B. Kato4, A. Dring1, H. E. Wichmann1,5,6, P. Deloukas2, M. Hrab de Angelis7,8, H. W. Mewes3,9, T. Illig1, T. D. Spector4, J. Adamski7,8, K. Suhre3,10, N. Soranzo2,4, C. Gieger1; 1 Institute of Epidemiology, Helmholtz Zentrum Muenchen, Neuherberg, Germany, 2Wellcome Trust Sanger Institute, Genome Campus, Hinxton, CB10 1HH, United Kingdom, 3Institute of Bioinformatics and Systems Biology, Helmholtz Zentrum Muenchen, Neuherberg, Germany, 4Department of Twin Research & Genetic Epidemology, Kings College London, London SE1 7EH, United Kingdom, 5Institute of Medical Informatics, Biometry and Epidemiology, Ludwig-Maximilians-Universitt, Munich, Germany, 6Klinikum Grosshadern, Munich, Germany, 7Institute of Experimental Genetics, Genome Analysis Center, Helmholtz Zentrum Muenchen, Neuherberg, Germany, 8Institute of Experimental Genetics, Life and Food Science Center Weihenstephan, Technische Universitt Muenchen, Munich, Germany, 9Department of Genomeoriented Bioinformatics, Life and Food Science Center Weihenstephan, Technische Universitt Muenchen, Munich, Germany, 10Faculty of Biology, Ludwig-Maximilians-Universitt, Munich, Germany.

Emerging technologies based on mass spectrometry and nuclear magnetic resonance enable the monitoring of hundreds of small metabolites from tissues or body fluids. Because metabolites change rapidly in response to physiologic perturbations, such metabolite concentrations provide a direct readout of the physiological state in the human body, leading to the discovery of novel proximal biomarkers of disease phenotypes. Furthermore, profiling of metabolites in relevant biological pathways can help elucidate the contribution of genetic variants underlying inherited variation in established risk factors. Among the major risk factors for coronary artery disease (CAD) and myocardial infarction (MI) are serum lipids, including total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), and triglycerides (TG). To dissect the effect of published genetic variants influencing serum lipid levels, we profiled 151 metabolites covering a biologically relevant panel of amino acids, sugars, acylcarnitines, and phospholipids in 1,797 participants from the KORA population (Germany) and replicated the results in 1,236 participants of the TwinsUK cohort. By analysing lipid concentrations in conjunction with genetic data and metabolite concentrations (and their ratios), we aim to identify cases where a genetic locus is associated with both a lipid and a metabolite concentration, which would provide new functional information about the underlying biological processes. We report here the initial results of this effort, as well as discussing methodological approaches to metabolite analyses. Among others, we identify associations of GCKR (glucokinase (hexokinase 4) regulator) variants with different ratios of plasmalogens and phosphatidylcholines (P = 3.2 10-8). C03.2 Identification of novel obesity loci by analysis of genomic structural variants
R. G. Walters1, L. J. M. Coin1, A. J. de Smith1, D. Meyre2, I. S. Farooqi3, P. Froguel1,2, A. I. F. Blakemore1; 1 Imperial College London, London, United Kingdom, 2CNRS 8090-Institute of Biology, Lille, France, 3Cambridge University, Cambridge, United Kingdom.

G. Trynka1, P. C. A. Dubois2, L. Franke1, *. Coeliac Disease Genetics Consortium3, R. McManus4, D. Barisani5, P. Deloukas6, J. C. Barrett6, P. Saavalainen7, D. A. Van Heel2, C. Wijmenga1; 1 University Medical Center Groningen, Groningen, Netherlands, 2Centre for Gastroenterology, Blizard Institute of Cell and Molecular Science, Barts and The London School of Medicine and Dentistry, London, United Kingdom, 3 University Medical Center Groningen and Barts and The London School of Medicine and Dentistry, Groningen, Netherlands, 4Departments of Clinical Medicine, Institute of Molecular Medicine, Trinity College Dublin, Dublin, Ireland, 5Department of Medical Sciences, University of Milan, Milan, Italy, 6 Wellcome Trust Sanger Institute, Hinxton, Cambridge, United Kingdom, 7 Department of Medical Genetics, Biomedicum Helsinki, University of Helsinki, Helsinki, Finland.

Coeliac disease is a complex, highly heritable trait. In our previous genome-wide association study (GWAS) and its follow-ups, we identified 13 non-HLA genomic risk regions for coeliac disease. Known variants, including HLA variants, explain around 35% of the heritability. We proposed that additional common genetic variants would underlie a further component of coeliac heritability. We performed a new GWAS in 4,533 coeliac cases and 10,750 controls from 4 populations of European descent. We integrated data from our first GWAS and expanded genomic coverage in the new samples to 523,749 SNPs passing quality controls. We further tested 131 SNPs in an additional cohort of 4,918 cases and 5,684 controls. Variants from 13 new regions showed genome-wide significance (Pcombined < 5 x 10-8), including regions containing TNFRSF14, RUNX3, CCR4, CD80, BACH2, THEMIS, ZMIZ1, ETS1, CIITA/SOCS1/CLEC16A, ICOSLG. A further 13 regions had suggestive association evidence (10-6<Pcombined<5x10-8, and/or Pfollow-up <0.01). Genes from most of these regions have an immune function, while newly identified associations to the TNFRSF14, RUNX3, ETS1 and THEMIS genes point to an unknown role for the thymic T cell selection pathway in the pathogenesis of coeliac disease. In a meta-analysis of expression quantitative traits in 1,469 whole blood samples, 20 out of the 38 (52.6%) tested loci had coeliac risk variants correlated (P < 0.0028, FDR 5%) with cis gene expression. Here we report multiple new common variants for coeliac disease and show that genetic determination of cis gene expression is a major mechanism by which these variants influence coeliac susceptibility. C03.4* Genetic variation in 22 loci influences QRS complex duration

Only a small fraction of the strong genetic contribution to obesity is explained by common variants identified from genome-wide association studies. To explore the contribution to obesity of rare variants with large effect, we are investigating the large genomic structural variants (GSVs) that are routinely found in patients with extreme phenotypes that include obesity as a key feature, and analysing these regions in case-control and population cohorts. Using this approach, we have shown that deletions of ~740kb at chromosome 16p11.2 - initially

A. Isaacs1, N. Sotoodenhia2, P. de Bakker3, M. Drr4, C. Newton-Cheh5, I. Nolte6, P. van der Harst7, M. Mller8, M. Eijgelsheim9, A. Alonso10, A. Hicks11, S. Padmanabhan12, C. Hayward13, A. Smith14, O. Polasek15, S. Giovannone16, I. Rudan17,18, J. F. Wilson18, P. Pramstaller11, D. Siscovick19, T. Wang5, V. Gudnason14, C. M. van Duijn1, S. B. Felix4, G. I. Fishman16, Y. Jamshidi20, B. Stricker9, N. J. Samani21, S. Kb22, D. E. Arking23, The QRS-GWAS Consortium; 1 Genetic Epidemiology Unit, Dep.of Epidemiology, Erasmus Univ.Medical Center, Rotterdam, Netherlands, 2Cardiovascular Health Research Unit, Dep. of Medicine, Univ.of Washington, Seattle, WA, United States, 3Program in Medical Population Genetics, Broad Institute of Harvard & MIT, Cambridge,

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MA, United States, 4Dep.of Internal Medicine B, Ernst-Moritz-Arndt-Univ., Greifswald, Germany, 5Center for Human Genetic Research, Cardiovascular Research Center, Massachusetts General Hospital, Boston, MA, United States, 6 Unit of Genetic Epidemiology and Bioinformatics, Dep.of Epidemiology, Univ. Medical Center Groningen, Groningen, Netherlands, 7Dep.of Cardiology, Univ. Medical Center Groningen, Groningen, Groningen, Netherlands, 8Institute of Epidemiology, Helmholtz Zentrum Mnchen, German Research Center for Environmental Health, Neuherberg, Germany, 9Dep.of Epidemiology, Erasmus Univ.Medical Center, Rotterdam, Netherlands, 10Div.of Epidemiology & Community Health, School of Public Health, Univ.of Minnesota, Minneapolis, MN, United States, 11Inst.of Genetic Medicine, European Academy Bolzano (EURAC) (Affiliated Inst. of Lbeck Univ., Germany), Bolzano, Italy, 12BHF Glasgow Cardiovascular Research Centre, Univ.of Glasgow, Glasgow, United Kingdom, 13MRC Human Genetics Unit, Inst.of Genetics and Molecular Medicine, Edinburgh, United Kingdom, 14Icelandic Heart Association Research Inst., Kopavogur, Iceland, 15Andrija Stampar School of Public Health, Univ. of Zagreb, Zagreb, Croatia, 16Div.of Cardiology, New York Univ.School of Medicine, New York, NY, United States, 17Croatian Centre for Global Health, Univ.of Split, Split, Croatia, 18Centre for Population Health Sciences, Univ.of Edinburgh, Edinburgh, United Kingdom, 19Cardiovascular Health Research Unit, Dep.of Medicine and Epidemiology, Univ.of Washington, Seattle, WA, United States, 20Div.of Clinical Developmental Sciences, St Georges Univ., London, United Kingdom, 21Dep.of Cardio Cardiovascular Sciences, Univ.of Leicester, Leicester, United Kingdom, 22Dep.of Medicine I, Klinikum Grosshadern, Munich, Germany, 23McKusick-Nathans Inst.of Genetic Medicine, Johns Hopkins Univ. School of Medicine, Baltimore, MD, United States

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sults from interactions between unknown environmental factors and alleles of many susceptibility loci across the genome. Recent investigations of the genetics of MS have resulted in important advances, driven largely by completion of the first genome-wide association scans (GWAS). To detect additional loci, we performed a GWAS in 882 Sardinian MS cases and 872 controls using 575,678 SNPs, genotyped with the Affymetrix 6.0 chip, that passed quality checks. Using imputation methods and haplotypes available from HapMap II, HapMap III and 1000 Genomes projects, we then imputed 6,031,588 SNPs and tested for association ~6.6 million variants. The strongest observed signal was on the HLA locus (p=1.4x10-20), at a SNP tag (r2=0.83) for the HLA-DRB1*0301 allele. We then ranked non-HLA SNPs based on their level of significance and proximity to functional candidate genes, and selected 9 SNPs with pvalue <1x10-5 for follow-up. Of those, one on chr3q13 was successfully confirmed in an independent sample of 1,775 MS cases and 2,005 controls (p=9.3x10-6), yielding to an overall pvalue of 1.6x10-10 (OR=1.40, C.I.=1.27-1.57). The most associated markers at this locus fall in the promoter of a gene that encodes a negative regulator of adaptive immune responses. Mice deficient for the ortholgoue are prone to experimental autoimmune encephalomyelitis, the animal model of MS but also spontaneously reject a variety of cancers. Hence, this gene appears critical for maintaining the balance between immune activation and tolerance. c03.6* Genome-wide association scan reveals major susceptibility locus in IL28B for both chronic Hepatitis C and for treatment failure

QRS complex duration, an ECG measurement reflecting ventricular depolarization, is associated with risk of heart failure, sudden death and mortality. The trait is known to have a heritable component (h2 ranging from 20 - 40%), however few genes influencing the trait have been identified to date. In the current study, a large-scale fixed-effects inverse variance weighted meta-analysis of GWAS data from 16 studies consisting of >40,000 individuals of European ancestry was performed. More than 2.5 million SNPs were imputed using the HapMap reference panel. Genomic control inflation factors were used to correct the test statistics at the individual population level as well as the final meta-analysis results. 612 SNPs achieved genome-wide significance levels (P<5e-8). These SNPs represented 30 independent association signals (r2<0.05) in 22 distinct genomic regions, of which only four were previously reported. These loci included genes in relevant cardiac conduction pathways, including sodium-channels (SCN10A and SCN5A), calcium handling (PLN/SLC35F1, STRN/HEATR5B, CASQ2, TKT/CACNA1D, and PRKCA), and transcription factors (NFIA, HAND1, TBX3, TBX5, TBX20, and KLF12), in addition to novel pathways (including kinase inhibition and growth-factor related genes). Multiple SNPs demonstrated pleiotropic effects with other ECG traits reflecting atrial conduction (PR interval) and myocardial repolarization (QT interval). Further, we showed experimentally that SCN10A, a gene in our most significantly associated region, is expressed preferentially in the mouse ventricular conduction system and affects mouse QRS interval duration. These analyses greatly expand our understanding of the genetic basis for variation in QRS duration and provide insights into the biology of cardiac conduction. c03.5* A genome-wide association scan in sardinians reveals a novel gene associated with multiple sclerosis

S. Sanna1, M. Pitzalis2, M. Zoledziewska2, I. Zara3, C. Sidore4, R. Murru5, M. B. Whalen4, F. Busonero1, A. Maschio1, G. Costa5, M. Pugliatti6, S. Traccis7, A. Angius4, M. Melis8, G. Rosati6, G. R. Abecasis9, M. Uda1, M. G. Marrosu5, D. Schlessinger10, F. Cucca1,2; 1 Istituto di Neurogenetica e Neurofarmacologia, Monserrato, Italy, 2Dipartimento di Scienze Biomediche, Universit di Sassari, Sassari, Italy, 3CRS4, Laboratorio di Bioinformatica, Parco tecnologico della Sardegna, Pula, Cagliari, Italy, 4 CRS4, Laboratorio di Genomica, Parco tecnologico della Sardegna, Pula, Cagliari, Italy, 5Centro Sclerosi Multipla, Dipartimento di Scienze Neurologiche e Cardiovascolari, Universit di Cagliari, Cagliari, Italy, 6Istituto di Neurologia Clinica, Universit di Sassari, Sassari, Italy, 7Presidio Ospedaliero, Divisione Neurologia, Sassari, Italy, 8AOBrotzu, S.C. Neurologia, Cagliari, Italy, 9 Department of Biostatistics, Center for Statistical Genetics, University of Michigan, Ann Arbor, MI, United States, 10Laboratory of Genetics, National Institute on Aging, Baltimore, MD, United States.

Z. Kutalik1,2, A. Rauch3, P. Descombes4, T. Cai5,6, J. di Iulio5, T. Mueller7, M. Bochud8, M. Battegay9, J. Borovicka10, S. Colombo5, A. Cerny11, J. Dufour12, H. Furrer3, M. Heim13, B. Hirschel14, R. Malinverni15, D. Moradpour16, B. Mllhaupt17, A. Witteck18, J. S. Beckmann1,19, T. Berg7, S. Bergmann1,2, F. Negro20, A. Telenti5, P. Bochud5,6; 1 Department of Medical Genetics, University of Lausanne, Lausanne, Switzerland, Lausanne, Switzerland, 2Swiss Institute of Bioinformatics, Lausanne, Switzerland, 3University Clinic of Infectious Diseases, University Hospital Bern and University of Bern, Bern, Switzerland, 4Genomics Platform, National Center of Competence in Research Frontiers in Genetics, University of Geneva, Geneva, Switzerland, 5Institute of Microbiology, University Hospital and University of Lausanne, Lausanne, Switzerland, 6Infectious Diseases Service, Department of Internal Medicine, University Hospital and University of Lausanne, Lausanne, Switzerland, 7Medical Clinic for Hepatology and Gastroenterology, Medical University Charit Campus, Virchow-Klinikum Berlin, Berlin, Germany, 8University Institute for Social and Preventive Medicine, University Hospital and University of Lausanne, Lausanne, Switzerland, 9Infectious Diseases and Infection Control Clinic, Department of Medicine, University Hospital Basel, Basel, Switzerland, 10Division of Gastroenterology, Canton Hospital St Gallen, St Gallen, Switzerland, 11Liver Unit, Clinica Luganese Moncucco, Lugano, Lugano, Switzerland, 12University Clinic of Visceral Surgery and Medicine, Inselspital, University of Bern, Bern, Switzerland, 13Division of Gastroenterology and Hepatology, University Hospital of Basel, Basel, Switzerland, 14Division of Infectious Diseases, University Hospital Geneva, Geneva, Switzerland, 15Pourtals Hospital, Neuchtel, Neuchatel, Switzerland, 16Division of Gastroenterology and Hepatology, University Hospital Lausanne, Lausanne, Switzerland, 17Division of Gastroenterology and Hepatology, University Hospital of Zurich, Zurich, Switzerland, 18Infectious Diseases and Infection Control Unit, Canton Hospital St Gallen, St Gallen, Switzerland, 19Service of Medical Genetics, Centre Hospitalier Universitaire Vaudois and University of Lausanne, Lausanne, Switzerland, 20Division of Gastroenterology and Hepatology, University Hospitals, Geneva, Geneva, Switzerland.

Multiple sclerosis (MS) is a multi-factorial neuroinflammatory and autoimmune disorder. A primary cause of disability in young adults, it re-

The hepatitis C virus (HCV) induces chronic infection in up to 80% of infected individuals, half of whom do not respond to therapy. We conducted a genome-wide association study to screen for host genetic determinants of HCV persistence and response to therapy. The analysis included 1362 hepatitis C infected individuals (448 of whom are co-infected with HIV): 1015 with chronic hepatitis C and 347 that spontaneously cleared the virus. Responses to pegylated interferon-alpha and ribavirin were assessed in 465 chronic hepatitis C patients. Associations between more than 2.5M single nucleotide polymorphisms (SNPs) and outcomes were computed using multivariate logistic regression. Chronic hepatitis C was found to be associated with SNPs in the IL28B locus, which encodes the antiviral cytokine interferon-lambda-3. The minor allele of the top hit rs8099917

Concurrent Sessions
was associated with progression to chronic HCV infection (OR=2.31, CI=1.74-3.06, P=6.07*10-9) explaining almost 3% of the phenotypic variance. The association was observed both in HCV mono-infected (OR=2.49, CI=1.64-3.79, P=1.96*10-5) and HCV/HIV co-infected individuals (OR=2.16, CI=1.47-3.18, P=8.24*10-5). Interestingly, SNP rs8099917 was also associated with failure to respond to therapy (OR=5.19, CI=2.90-9.30, P= 3.11*10-8), with the strongest effects in patients with HCV genotypes 1 or 4, where more than 11% of the variance in response was explained by this genotype alone. Re-sequencing of IL28B identified distinct haplotypes that were associated with the clinical phenotype. The association of the IL28B locus with natural and treatment-associated control of HCV indicates the importance of innate immunity and interferon-lambda-3 in the pathogenesis of HCV infection. C04.1 Identification of novel deafness genes by homozygosity mapping in Dutch families

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sive, but the rate of decline is variable within and between families. The observation of recurrences in siblings in some families suggested that the condition was probably autosomal recessive, however the variable age of onset and clinical course raised the possibility that it may be aetiologically heterogeneous. We identified a candidate gene, C20orf54 by studying a consanguineous family with multiple affected individuals and subsequently demonstrated that mutations in this gene were the cause of disease in other, unrelated families. c04.3* the microRNA miR-204 is required for vertebrate eye development

I. Conte1, S. Carrella1, R. Avellino1, M. Karali1, R. Marco-Ferreres2, P. Bovolenta2, S. Banfi1; 1 TIGEM, Naples, Italy, 2Departamento de Neurobiologa Molecular Celular y del Desarrollo, Instituto Cajal, CSIC, and CIBER de Enfermedades Raras (CIBERER), Madrid, Spain.

H. Kremer1,2, J. Oostrik3, P. L. M. Huygen2, J. A. Veltman1,4, L. H. Hoefsloot4, C. W. R. J. Cremers5,2, H. P. M. Kunst5,2, R. J. C. Admiraal5,2, M. Schraders1,2; 1 Nijmegen Center for Molecular Life Sciences, Nijmegen, Netherlands, 2Dept Otorhinolaryngology, Radboud University Nijmegen Medical Centre, Nijmegen, Netherlands, 3Dept. Otorhinolaryngology, Radboud University Nijmegen Medical Centre, Nijmegen, Netherlands, 4Dept Human Genetics, Radboud University Nijmegen Medical Centre, Nijmegen, Netherlands, 5Donders Institute for Brain, Cognition and Behaviour, Radboud University Nijmegen, Nijmegen, Netherlands.

Locus heterogeneity of autosomal recessive nonsyndromic hearing loss (arNSHI) is large. So far about 30 genes have been identified but at least 50 genes await identification. Novel arNSHI loci are generally determined in large consanguineous families and are often large (1030 Mb). Although next generation sequencing enables the simultaneous sequencing of exons and regulatory regions of all genes within a region, strategies to delimit the critical region and intelligent candidate gene selection remain attractive for disease gene identification. Although the majority of the Dutch population is regarded to be mixed there are quite a number of regions with limited migration mainly until about 1950. Therefore, we followed a strategy of homozygosity mapping with high density SNP arrays in 125 patients with putative arNSHI from 77 families to delimited the critical region of known deafness loci and/or identify novel loci. Homozygous regions smaller than 1 Mb were not further investigated. We delimited the critical region for DFNB25 to an 0.8 Mb interval harbouring exons of two genes. Also, we obtained indications for a novel locus, now DFNB84, of 3.2 Mb with 11 known and predicted genes. Mutation analysis of all exons in the DFNB25 interval and selected candidate genes in the novel locus revealed putatively pathogenic mutations in GRXCR1 and PTPRQ. Vestibular dysfunction can be associated with the hearing loss for both genes. Our results indicate that the demographic structure of the Dutch population is favourable for this method which is likely to be the case in more European countries. c04.2 Brown-Vialetto-Van Laere syndrome, a ponto-bulbar palsy with deafness, is caused by mutation in C20orf
D. J. Josifova1, P. Green2, M. Wiseman2, Y. J. Crow3, H. Houlden4, S. Riphagen5, J. Lin5, F. Raymond6, A. Childs7, E. Sheridan8, S. Edwards2; 1 Guys Hospital, London, United Kingdom, 2Kings College, London, United Kingdom, 3Manchester Academic Health Science Centre, St. Marys Hospital, Manchester, United Kingdom, 4Institute of Neurology, London, United Kingdom, 5 Evelina Childrens Hospital, St. Thomas Hospital, London, United Kingdom, 6 Cambridge Institute for Medical Research, Addenbrooks Hospital, Cambridge, United Kingdom, 7The General Infirmary at Leeds, Leeds, United Kingdom, 8 , Leeds Institute of Molecular Medicine, St Jamess Hospital, Leeds, United Kingdom.

The functional role of specific microRNAs in controlling the morphogenetic and cell differentiation events involved in normal eye development in vertebrates is still largely unknown. Here we show that a single microRNA, miR-204, is capable to regulate multiple aspects of eye development in medaka fish. Targeted ablation of miR-204 function by morpholino injections in medaka determined a severe eye phenotype characterized by microphthalmia, aberrant lens formation, incorrect retinal cell differentiation and coloboma. Through a variety of in vitro and in vivo approaches, we found that Meis2 is a key target of miR204 and plays a pivotal role in the generation of this phenotype via the regulation of the Pax6 pathway. These data demonstrate for the first time that a specific microRNA is involved in the regulation of basic processes underlying eye development and open new avenues on a better comprehension of the pathogenetic mechanisms underlying eye developmental disorders. C04.4* Phenotypic modifiers of DJ1
S. Jain, P. Heutink; CNCR, Amsterdam, Netherlands.

Parkinsons disease (PD) is a slow progressing neurodegenerative disease with devastating clinical symptoms. Current treatments are only symptomatic which ultimately result in debilitating side effects thus there is an urgent need to develop therapeutics which stop and reverse disease progression. None of the 13 loci implicated in the PD pathogenesis represent viable drug targets at present. In addition these genes have suggested that disruption of a myriad of molecular pathways (e.g. ubiquitin and proteasome, mitochondrial function and protein misfolding) can lead to the degeneration of the substantia nigra. It remains unclear how mutation of these genes and disruption of these pathways lead to PD and thus complicates the process for drug discovery. To address these issues, we conducted a high content screening to interactors of DJ1, a gene mutated in PD to determine if any of them is capable of affecting DJ1 function. Thus far we have been able to identify several genes which are able to rescue DJ1 deficits on cell viability such as PPP2R2C and PSF and other genes which are able to enhance the loss of DJ1 such as 4E-BP. 4E-BP is a translational inhibitor to which many drugs are available thus the administration of 4E-BP activators, such as rapamycin may protect cells from apoptosis and thus an effective treatment to prevent or delay the onset of disease. Using this approach we have been able to construct a detailed molecular pathway of the proteins that are involved in the function of DJ1 and identify additional therapeutic targets. c04.5 miR-135b regulates two transcriptional cofactors, Pc4 and Psip1, in the mammalian inner ear, identified using an integrative transcriptomics and proteomic approach
T. Elkan1, R. Hertzano1, I. Ulitsky2, R. Elkon1,2, M. Irmler3, R. Shamir2, J. Beckers3, K. B. Avraham1; 1 Department of Human Molecular Genetics and Biochemistry, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel, 2Blavatnik School of Computer Science, Tel Aviv University, Tel Aviv, Israel, 3Institute of Experimental Genetics, Helmholtz Zentrum Mnchen, Neuherberg, Germany.

Brown-Vialetto-Van Laere syndrome is a rare neurological disorder first reported by Brown in 1894 as familial amyotrophic lateral sclerosis with onset in infancy. Following the reports by Vialetto, in 1936 and Van Laere, in 1966, the name, BVVLS was adopted. The key features are progressive ponto-bulbar palsy and bilateral sensorineural deafness. The disease usually presents with VII, IX, X, XI and XII cranial nerve palsies, which develop in a previously healthy individual. A complex neurological phenotype with a mixed picture of upper and lower motor neuron involvement reminiscent of amyotrophic lateral sclerosis evolves with disease progression. The course is invariably progres-

MicroRNAs (miRNAs) are 17-24 nucleotide-long non-coding RNAs processed from transcripts of endogenous genes that function through the RNA interference (RNAi) pathway. miRNAs regulate gene expression by inducing degradation of mRNA of target genes and by inhib-

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iting translation. Their relevance to the inner ear has recently been emphasized by the discovery of miRNA mutations leading to deafness in humans and mice. We integrated a comparative transcriptomic and proteomic analyses and a miRNA screen of early post-natal cochlear and vestibular sensory epithelia derived from mice, with sequencebased predictions, to efficiently identify functional miRNAs and their targets. We identified PC4 and Psip1, two transcriptional cofactors that interact with one another, as targets for miR-135b in the inner ear hair cells. PC4, Activated RNA polymerase II transcription cofactor 4, or Sub1, mediates functional interactions between upstream activators and general transcriptional machinery. Psip1, also known as PC4and SF-2 interacting protein or Lens epithelium-derived growth factor (Ledgf), is involved in transcriptional regulation of stress related genes, mRNA splicing, and cell survival. In order to prove the interaction between the miRNA and its target proteins the miRNA was silenced or over-expressed in cell line and the protein levels were studied using semi-quantitative western blot analysis. Our current work focuses on how miR-135b regulation affects downstream pathways in the inner ear. c04.6* Olfactory Expression of mutant A30P alpha-synuclein in conditional mouse Brain: implications for Early stage of Parkinsons Disease
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Dept of Biochemistry, University of Pavia, Pavia, Italy, 3Genetics Unit, Dept of Pediatrics, Hacettepe Univ faculty of medicine, Ankara, Turkey, 4Dept of Paediatrics, Faculty of medicine and health sciences, Al Ain, United Arab Emirates, 5Dept of Medical Genetics, Univ medical Center, Utrecht, Netherlands, 6Jean Verdier hospital, Bondy, France, 7Dept of Medical Genetics, UNICAMP, Sao Paulo, Brazil, 8Division of Paediatric Orthopedics, Orthopedic Hospital Speising, Vienna, Austria, 9Dept of Medical Genetics, Antwerp University Hospital, Antwerp, Belgium, 10Dept of Pediatrics and Child Health, Dunedin School of Medicine, University of Otago, Dunedin, New Zealand, 11 Genetics Research Center, Univ of Social Welfare and Rehabilitation Sciences, Teheran, Islamic Republic of Iran, 12Centre for Pediatrics and Adolescent Medicine, Freiburg Univ Hospital, Freiburg, Germany, 13Dept of Pathology, Campus Bio-Medico University, Roma, Italy.

S. Nuber1, E. Petrasch-Parwez2, O. Arias-Carrion3, C. Wurst1, F. N. Gellerich4, Z. Gizatullina5, M. Fendt6, H. Nguyen7, S. von Hrsten8, P. Teismann9, J. B. Schulz10, T. P. Velavan11, T. Schmidt1, J. Boy1, I. Schmitt12, G. U. Hglinger13, J. Winkler14, O. Riess1; 1 Medical Genetics, Univ. of Tbingen, Germany, 2Institute of Neuroanatomy and Molecular Brain Research, Univ. of Bochum, Germany, 3Inst. of Experimental Neurology, Philipps University of Marburg, Germany, 4KeyNeurotec, ZENIT Technology Park, Magdeburg, Germany, 5KeyNeurotek, ZENIT Technology Park, Magdeburg, Germany, 6Novartis, Basel, Switzerland, 7Medical Genetics, Univ.of Tbingen, Germany, 8Department of Experimental Therapy, Univ. of Erlangen, Germany, 9Inst. of Medical Sciences, University of Aberdeen, United Kingdom, 10RWTH, Univ. of Aachen, Germany, 11Inst. for Tropical Medicine, Univ. of Tbingen, Germany, 12Clinic for Neurology, Univ. of Bonn, Germany, 13 Experimental Neurology, Philipps University of Marburg, Germany, 14Division of Molecular Neurology, Univ, of Erlangen, Germany.

Growing evidence suggests that cognitive and psychiatric deficits precede motor impairment in Parkinsons disease (PD). In this premotor stage neuropathology is detectable in the olfactory bulb and smell deficiency is found in about 90% of PD patients. To explore the impact and reversibility of early pathological stages, we analyzed conditional transgenic mice, expressing high levels of human [A30P]alpha-synuclein limited to the olfactory bulb. Further, as alpha-synuclein positive olfactorial lesions of early PD-stages appears not to advance to non-olfactorial cortical areas, which are increasingly affected at later stages, their contribution to disease progression is unknown. Thus, the question raise of whether (i) mutated alpha-synuclein is able to induce olfactorial pathology,(ii) these alterations influence non-olfactorial brain structures and (iii) neurological dysfunction is reversible after ceasing expression of transgene alpha-synuclein. Thus, we generated and characterized a tet-off conditional mouse model expressing human [A30P]alpha-synuclein in the olfactory bulb. We found that overexpressing of mutated [A30P]alpha-synuclein led to a downregulation of dopamine neurotransmission in the olfactory bulb which was paralleled by hyperactivity and decreased anxiety in these mice. We further detected upregulation of neurotransmitter content in striatum and substantia nigra and mitochondrial dysfunction in non-olfactory brain regions, both of which could be reversed in old-aged mice. Perspective: Using this regulatable transgenic mouse model, we may model and explore in detail impact of olfactory alpha-synucleinopathy on other brain regions; this model is also a useful tool to study early intervention strategies, which may halt or even reverse the underlying disease process in PD. c05.1 cANt1 mutations in Desbuquois dysplasia are responsible for a defect in proteoglycan synthesis.
C. Huber1, A. Rossi2, M. Bertoli1, M. Fradin1, M. Le Merrer1, Y. Alanay3, L. I. Al Gazali4, M. G. E. M. Ausems5, P. Bitoun6, D. P. Cavalcanti7, A. Krebs8, G. Mortier9, S. P. Robertson10, Y. Shafeghati11, A. Superti-Furga12, A. O. Muda13, C. Le Goff1, A. Munnich1, V. Cormier-Daire1; 1 INSERM U781, Paris Descartes University, Necker Hospital, Paris, France,

Desbuquois dysplasia is an autosomal recessive chondrodysplasia characterized by short stature, joint laxity, scoliosis and advanced carpal ossification with a delta phalanx. Studying 9 Desbuquois families, we identified 7 distinct mutations in the Calcium-Activated Nucleotidase 1 gene (CANT1) which encodes a soluble UDP-preferring nucleotidase. Among the 7 mutations, 4 were nonsense and 3 were missense mutations, located in the region encoding the 7th nucleotidase conserved region and changing arginine at position 300 in 5/9 families. All children presented with characteristic skeletal manifestations. However, an early death due to cardio-respiratory failure was observed in the 4 children with non sense mutations. The function of CANT1 is unknown. Using RT-PCR analysis, we observed a specific expression in chondrocytes. We also found electron-dense material within distended rough endoplasmic reticulum in Desbuquois patient fibroblasts. Finally, Desbuquois dysplasia shares phenotypic features with Diastrophic dysplasia and recessive Larsen syndrome, which are both due to a defect in proteoglycan sulfation, the final step of proteoglycan synthesis. To test whether CANT1 deficiency interferes with the availability of UDP-sugars needed for proteoglycan synthesis, fibroblasts from two Desbuquois patients and four controls were double labeled with [35S]sulfate and [3H]glucosamine. Surprisingly, in the patient cells glycosaminoglycan (GAG) synthesis was almost normal under basal conditions when compared to controls, but significant reduced GAG synthesis was observed in presence of -Dxyloside, a compound that increases GAG synthesis acting as a chain initiator. These data suggest that CANT1 plays a role in proteoglycan metabolism and supports its involvement in the rate of GAG synthesis. c05.2* Disruption of the Podosome Adaptor Protein tKs4 (SHPXD2B) causes Frank-ter Haar syndrome

Z. Iqbal1, P. Cejudo-Martin2, A. De Brouwer1, B. Van der Zwaag3, P. RuizLozano2, M. Cecilia Scimia2, J. D. Lindsey4, R. Weinreb4, B. Albrecht5, A. Megarbane6, Y. Alanay7, Z. Ben-Neriah8, M. Amenduni9, R. Artuso9, J. A. Veltman1, E. Van Beusekom1, A. Oudakker1,10, J. Luis Millan2, R. Hannekam11,12, B. Hamel1, S. A. Courtneidge2, H. van Bokhoven1,10; 1 Department of Human Genetics, Nijmegen Centre for Molecular Life Sciences, Radboud University Nijmegen Medical Center, Nijmegen, Netherlands, 2 Burnham Institute for Medical Research, 10901 N. Torrey Pines Road, La Jolla, CA 92037, USA, La Jolla, CA, United States, 3Department of Neuroscience and Pharmacology. University Medical Center Utrecht, Utrecht, Netherlands, 4Hamilton Glaucoma Center, University of California at San Diego, 9500 Gilman Drive, La Jolla, CA 92093, USA., La Jolla, CA, United States, 5Institut fr Humangenetik, Universittsklinikum, Universitt DuisburgEssen, Germany., Duisburg-Essen, Germany, 6Medical Genetics Unit, Saint Joseph University, Beirut, Lebanon., Beirut, Lebanon, 7Pediatric Genetics Unit, Department of Pediatrics, Hacettepe University Faculty of Medicine,, Ankara, Turkey, 8Center for Human Genetics, Hadassah Medical Center, Hebrew University of Jerusalem,, Jerusalem, Israel, 9Medical Genetics, Department of Molecular Biology, University of Siena, Siena, Italy, 10Department of Cognitive Neurosciences, Donders Institute for Brain, Cognition and Behaviour, Radboud University Nijmegen, Nijmegen, Netherlands, 11Department of Pediatrics, Academic Medical Centre, Amsterdam, Netherlands, 12Institute of Child Health, Great Ormond Street Hospital for Children, UCL, London, United Kingdom.

Frank-Ter Haar syndrome (FTHS), also known as Ter Haar syndrome, is an autosomal recessive disorder characterized by skeletal, cardiovascular and eye abnormalities, such as increased intraocular pressure, prominent eyes and hypertelorism. We have conducted homozygosity mapping on patients representing twelve FTHS families. A locus on chromosome 5q35.1 was identified for which patients from

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ten families shared homozygosity. For one family, a homozygous deletion mapped exactly to the smallest region of overlapping homozygosity, which contained a single gene, SH3PXD2B. This gene encodes the TKS4 protein, a PX and SH3 domain-containing adaptor protein and Src substrate. This protein was recently shown to be involved in the formation of actin rich membrane protrusions called podosomes or invadopodia, which coordinate pericellular proteolysis with cell migration. Mice lacking Tks4 also show pronounced skeletal, eye and cardiac abnormalities and phenocopied the majority of the defects associated with FTHS. These findings establish a role for TKS4 in FTHS and embryonic development. Mutation analysis revealed five different homozygous mutations in SH3PXD2B in seven FTHS families. No SH3PXD2B mutations were detected in six other FTHS families, demonstrating the genetic heterogeneity of this condition. Interestingly however, dermal fibroblasts from one of the individuals without an SH3PXD2B mutation nevertheless expressed lower levels of the TKS4 protein, suggesting a common mechanism underlying disease causation.This is the first time that a developmental disorder is caused by the defect in podosomes. Further study on the role of podosomes may open a new horizon in identifying the genetic defects in related developmental disorders. c05.3* PTHLH deletion and point mutations are associated with Brachydactyly type E (BDE)
E. Klopocki1, B. P. Hennig1, K. Dathe1, R. Koll1, T. de Ravel2, E. Baten3, E. Blom4, Y. Gillerot5, J. F. W. Weigel6, G. Krger7, O. Hiort8, P. Seemann9, S. Mundlos1,10; 1 Charite Universittsmedizin Berlin, Berlin, Germany, 2Centre for Human Genetics University Hospital Leuven, Leuven, Belgium, 3AZ Saint Lucas, Brugge, Belgium, 4Department of Clinical Genetics, Maastricht University Center, Maastricht, Netherlands, 5Centre de Gntique Humaine, Cliniques Universitaires St Luc, Brussels, Belgium, 6University Childrens Hospital, Leipzig, Germany, 7Abteilung Medizinische Genetik, Universittsklinikum Rostock, Rostock, Germany, 8Departments of Pediatrics, University of Lbeck, Lbeck, Germany, 9Berlin-Brandenburg Center for Regenerative Therapies (BCRT), Berlin, Germany, 10Max-Planck-Institut fr Molekulare Genetik, Berlin, Germany.

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Medicine, Ankara, Turkey, 5Dept Pediatrics, Akdeniz Univ School of Medicine, Antalya, Turkey, 6Dept Medical Genetics, National Institute of Health, Rabat, Morocco, 7Dept Radiology, Erasmus MC, Rotterdam, Netherlands, 8Centre for Pediatrics, Univ of Freiburg, Freiburg, Germany.

Spondylo-megaepiphyseal-metaphyseal dysplasia (SMMD) is a rare autosomal recessive skeletal dysplasia with only a few cases reported in the literature. Affected individuals have a disproportionate short stature with a short neck and trunk and relatively long limbs that may show flexion contractures of the distal joints. The most remarkable radiographic features are the delayed and impaired ossification of the vertebral bodies as well as the presence of large epiphyseal ossification centers and wide growth plates in the long tubular bones. Genome wide homozygosity mapping followed by a candidate gene approach resulted in the elucidation of the genetic cause in three new consanguineous families with SMMD. Each proband was homozygous for a different inactivating mutation (c.336_337delGGinsT; c.337dupG; c.104_110delCGCCCG) in NKX3-2, a homeobox-containing gene located on chromosome 4p15.33. Expression studies in skin fibroblasts showed partial nonsense mediated decay of the mutant transcripts and upregulation of mutant NKX3-2 mRNA compared to control, the latter suggesting a disturbed autoregulatory loop. We observed the highest expression of NKX3-2 in chondrocytes and gut. In contrast to previous reports we were also able to demonstrate NKX3-2 expression in the brain. Striking similarities were found when comparing the vertebral ossification defects in SMMD patients with those observed in the Nkx3-2 null mice. Distinguishing features were the asplenia found in the mutant mice and the radiographic abnormalities in the limbs only observed in SMMD patients. This study illustrates that NKX3-2 plays an important role in endochondral ossification of both the axial and appendicular human skeleton. c05.5 copy Number and sequence Variants in FREM1 are Associated With an increased Risk of isolated metopic craniosynostosis in Humans and mice
L. E. L. M. Vissers1, T. C. Cox2, I. M. Janssen1, K. M. Short3, I. Smyth4, F. Jehee5, G. Yagnik6, S. A. Boyadjiev6, C. Marcelis1, P. J. Anderson7, M. L. Cunningham2, M. Passos-Bueno5, J. A. Veltman1, M. F. Buckley1, T. Roscioli1; 1 Department of Human Genetics, Nijmegen, Netherlands, 2Division of Craniofacial Medicine, University of Washington, Seattle, WA, United States, 3 Cutaneous Developmental Biology Lab, Monash University, Melbourne, Australia, 4Cutaneous Developmental Biology Lab, Monash University, Melbourne, Australia, 5Centro de Estudos do Genoma Humano, Universidade de So Paulo, So Paulo, Brazil, 6Section of Genetics, Department of Pediatrics, University of California, Davis, Sacramento, CA, United States, 7 Australian Craniofacial Unit, Adelaide, Australia.

Brachydactylies are a family of limb malformations characterized by short hands/feet due to aplastic/hypoplastic skeletal elements. Autosomal dominant brachydactyly type E (BDE, MIM 113300) is characterized by shortening of metacarpals/metatarsals and/or phalanges. Here we describe a novel disease gene for BDE in five unrelated families. Initially we detected a microdeletion encompassing PTHLH, the gene coding for parathyroid hormone related protein (PTHRP) in a pedigree with BDE, short stature and learning disabilities. PTHRP regulates the balance between chondrocyte proliferation and hypertrophic differentiation during endochondral bone development. Pthrp-/- mice show short limbed dwarfism due to premature differentiation of chondrocytes. Therefore we screened a cohort of individuals with BDE and short stature for mutations in PTHLH. We identified two missense (p.L44P and p.L60P), a nonstop (p.X178WextX*54), and a nonsense (p.K120X) mutation. Functional testing of the L60P missense mutation in chicken micromass culture showed an earlier differentiation of chondrocytes compared to wildtype which indicates that the missense mutation results in a loss of function. Since mutations of the primary mediator of PTHRP/PTH receptor signaling, GNAS1, are associated with Albright Hereditary Osteodystrophy (AHO) which includes a skeletal phenotype strikingly similar to the BDE phenotype we conclude that the IHH/PTHRP pathway is of particular importance for cartilage differentiation and growth in the metacarpals. These results support the concept of molecular disease families based on phenotypic similarities and interacting signaling pathways. Taken together, mutations in PTHLH result in a specific type of skeletal disease which we suggest to name BDE with short stature, PTHLH type. c05.4 Homozygous inactivating mutations in the NKX3-2 gene result in spondylo-megaepiphyseal-metaphyseal dysplasia

G. R. Mortier1, M. Simon2, A. Dheedene3, Y. Alanay4, E. Mihci5, L. Rifai6, A. Sefiani6, Y. van Bever2, M. Meradji7, A. Superti-Furga8, J. Hellemans3; 1 Dept Medical Genetics, Antwerp Univ Hosp, Antwerp, Belgium, 2Dept Clinical Genetics, Erasmus MC, Rotterdam, Netherlands, 3Dept Medical Genetics, Ghent Univ Hosp, Ghent, Belgium, 4Dept Pediatrics, Hacettepe Univ School of

Metopic craniosynostosis (MC), the premature fusion of the frontal bone primordia, results in trigonocephaly and has a frequency of 1:10,000 live births. One form of MC is associated with monosomy for an 8Mb interval of chromosome 9p22.3 (OMIM 158170). Features include mental retardation, trigonocephaly and midface hypoplasia. Overlapping copy number variants (CNVs) have been identified which include the FREM1 gene. We wished to investigate the role of FREM1 in more than 100 people with MC ascertained through an international craniofacial consortium. The presence of 9p CNVs was assessed by microarrays and FREM1 was screened for mutations by sequencing and MLPA. Frem1 knockout mice were also imaged with microCT to determine if craniofacial anomalies consistent with a human MC phenotype. Two de novo CNVs involving FREM1 and 3 human FREM1 alleles (one de novo) were identified. The two CNVs interrupt the FREM1 coding sequence, likely resulting in structural abnormalities of the FREM1 protein. Evidence that a p.Glu1500Val mutation may be involved in MC includes that it is de novo in a single family and is present in a second family with a range of craniofacial abnormalities. Craniofacial imaging of homozygous Frem1 knock-out mice demonstrated craniofacial deformities consistent with MC. Taken together, these findings account for a significant percentage of a uni-sutural craniosynostosis and are consistent with FREM1 variants increasing the risk of MC. We recommend that all children with MC should have 9p CNV investigations and consideration be given to FREM1 mutation analysis.

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c05.6 cranioectodermal dysplasia is a ciliary disorder caused by defects in the iFt122 gene

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this cohort. Genetic disorders were found in 110 children. Six children presented with Beckwith-Wiedemann syndrome (0.04% vs 0.007% expected) and 5 with a retinoblastoma (0.03% vs 0.006%), both diseases influenced by epigenetics. More investigations will highlight detailed informations about factors impacting on the occurrence of these abnormalities, as parental patterns, indications or ART technics. c06.2 the origin of de novo chromosome deletions identified by array cGH
S. Thomas, C. Sibbons, H. Eisenhauer, J. Crolla, P. Jacobs; Wessex Regional Genetics Laboratory, Salisbury, United Kingdom.

J. Walczak-Sztulpa1, J. Eggenschwiler2, D. Osborn3, D. A. Brown2, F. Emma4, C. Klingenberg5, R. C. Hennekam6, G. Torre7, M. Garshasbi1, A. Tzschach1, M. Szczepanska8, M. Krawczynski9, J. Zachwieja10, D. Zwolinska11, P. Beales3, H. Ropers1, A. Latos-Bielenska12, A. W. Kuss1; 1 Max Planck Institute for Molecular Genetics, Berlin, Germany, 2Department of Molecular Biology, Princeton University, Princeton, NJ, United States, 3 Molecular Medicine Unit, University College London (UCL) Institute of Child Health, London, United Kingdom, 4Division of Nephrology and Dialysis, Childrens Hospital and Research Institute Bambino Gesu, Rome, Italy, 5 Department of Paediatrics, University Hospital of North-Norway and University of Troms, Troms, Norway, 6Institute of Child Health, Great Ormond Street Hospital for Children, University College London, London, United Kingdom, 7 Division of Hepato-Gastroenterology, Childrens Hospital and Research Institute Bambino Gesu, Rome, Italy, 8Department of Gynecology and Obstetrics, Division of Reproduction, Poznan University of Medical Sciences, Poznan, Poland, 9Department of Gastroenterology and Metabolism, Poznan University of Medical Sciences, Poznan, Poland, 10Department of Paediatric Cardiology and Nephrology, Poznan University of Medical Sciences, Poznan, Poland, 11Department and Clinic of Paediatric Nephrology, Wroclaw Medical University, Wroclaw, Poland, 12Department of Medical Genetics, Poznan University of Medical Sciences, Poznan, Poland.

Cranioectodermal dysplasia (CED, Sensenbrenner syndrome, OMIM 218330) is characterized by craniofacial, skeletal, and ectodermal abnormalities. Most cases are sporadic, but a few familial ones support an autosomal recessive inheritance pattern. We collected 13 CED patients from 12 independent families. In two families the patients had consanguineous parents, and in one of these, two siblings were affected, permitting linkage analysis and homozygosity mapping. This revealed a single region of homozygosity with a significant LOD score (3.57) on chromosome 3q21-3q24. By investigating candidate genes from this interval we found a homozygous missense mutation in exon 15 (V->G) of the IFT122 gene that co-segregated with the disease. In addition we identified another homozygous missense change (S->F) in exon 11 if IFT122 in the patient from the second consanguineous family, and found compound heterozygosity for two different IFT122 mutations (a splice site change in intron 6 and a missense change in exon 1) in a sporadic patient. All changes were absent in 340 control chromosomes. The IFT122 protein is a component of the retrograde intraflagellar transport and important for in the assembly and maintenance of eukaryotic cilia. We therefore investigated the primary cilia in patient fibroblasts and found significantly reduced cilia frequency and length in patient cells as compared to controls. We next transiently knocked down ift122 in zebrafish embryos and observed a characteristic ciliary phenotype, confirming that CED is a ciliary disorder and suggesting that the causative mutations in the unresolved cases are most likely to affect primary cilia function as well. c06.1 is there an increased risk of congenital malformations after Assisted Reproductive technologies (ARt)? Results of a French cohort composed of 15 162 children

Among de novo cytogenetically visible chromosome imbalances with non-recurrent breakpoints, there is an overall excess of paternal cases: this effect is genome wide but varies according to the type of abnormality and the chromosomal location. These imbalances are likely to arise meiotically and are not associated with increased parental age. In contrast, while non-recurrent balanced translocations are also predominantly paternal in origin, they display a significantly elevated paternal age suggesting they may arise during mitosis in male germ cells prior to meiosis. The use of array CGH provides two significant advantages in the study of chromosome abnormalities: it allows the aetiology of very small imbalances to be investigated and defines precisely the size of an imbalance thus giving information on the sequence composition around the breakpoints. We have investigated the parental origin of 37 deletions identified or characterised by array CGH using the Agilent 44K platform. Twenty seven (73%) were paternal in origin and 10 maternal (27%). Analysis of the breakpoint intervals showed that one of the maternal cases and five of the paternal cases contained segmental duplications suggesting formation by Non-Allelic Homologous Recombination. The average size of the paternal deletions was 7.3Mb compared with 3.1Mb for the maternal cases. These results confirm the parental origin bias, although among deletions below 6Mb the excess of paternal cases was less pronounced. We will investigate the origin of further cases including duplications and complex abnormalities with multiple breakpoints and also look for any association with increased parental age. c06.3 Novel insights into the pathogenesis of common aneuploidies using genomic analysis of cell-free amniotic fluid
D. W. Bianchi1, K. L. Johnson1, K. Koide1, L. Massingham1, J. L. Maron1, D. Slonim2; 1 Tufts Medical Center, Boston, MA, United States, 2Tufts University, Medford, MA, United States.

G. B. Viot1, S. Epelboin2, F. Olivennes3, S. Follow-Up AMP vigilance4; 1 Maternit Port Royal, Paris, France, 2Maternit Saint Vincent de Paul, Paris, France, 3Clinique de la Muette, Paris, France, 4Poissy, Poissy, France.

The aims of this study is to estimate the risk of major or minor congenital malformations and genetic disorders among children born after in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) in 33 French centers registered for ART. The survey was prospective from 2003 to 2007. The cohort was composed of 15 162 infants recruited from birth; medical data were collected regularly from birth to the age 60 months. Questionnaires were fulfilled by the paediatrician and the parents and 98% of them were exploitable. The prevalence of each malformation identified among this cohort was compared to the data of national registers. A major congenital malformation was found in 4.24% children (vs 2-3% expected). This higher rate was partly due to an excess of heart diseases (0.45% vs 0.25%) and malformations in the urogenital system [uropathy : 1.25% (vs 0.08-0.65%), hypospadias : 0.37% (vs 0.29%)]. Among the minor malformations, a 5 times higher frequency of angioma was reported with a totally skewed sex ratio (262 girls/103 boys). Surprisingly, the average age for parents of malformed children at the time of the conception was not statically different from the all parents of

Background: As a novel means of identifying pathophysiologic changes in fetuses with common aneuploidies, we characterized developmental gene expression using cell-free mRNA in residual second trimester amniotic fluid (AF) supernatant samples. Methods: RNA was extracted from AF in fetuses with trisomy 21 [T21] (n=7), trisomy 18 [T18] (n=5), and euploid controls (n=13). cDNA synthesis and biotin labeling were performed prior to hybridization to Affymetrix U133 plus 2.0 arrays. Initial analysis was done using the Affymetrix Gene Chip Microarray Suite 5.0, followed by comparative t-tests and Benjamini-Hochberg adjustment. Differentially-expressed genes were further examined by the Database for Annotation, Visualization, and Integrated Discovery (DAVID) and Ingenuity. Results: In T21 we identified 414 differentially-expressed genes vs. controls. Only 5/414 genes mapped to chromosome 21. In T18, only 7/356 differentially-expressed genes were on chromosome 18. Heat map and functional analyses of genes not on 18 or 21 showed a consistent pattern unique for each aneuploidy that significantly differed from euploidy. Only 6 differentially-expressed transcripts were common to both aneuploidies. T21 samples showed significant oxidative stress and disruptions in ion transport, G-protein signaling, immune response, and circulatory system function. T18 showed down-regulation of the endocrine system and up-regulation of lipid metabolism. Conclusions: Our results question the conventional wisdom that the pathophysiology of aneuploidy is due to a gene dosage effect, as the molecular abnormalities observed here are predominantly produced by genes on chromosomes other than 18 or 21. This discovery-driven genomic approach using discarded material suggests new avenues to

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further understand abnormal fetal development. c06.4* Frequencies of 15q11-q13, 7q11.23 and 22q11.2 deletions and duplications in spermatozoa from Prader-Willi syndrome fathers.
. Molina, E. Anton, F. Vidal, J. Blanco; Unitat de Biologia Cellular (Facultat de Biocincies). Universitat Autnoma de Barcelona, Cerdanyola del Valls, Spain.

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c06.6* Prenatal anomalies in a cohort of 40 Noonan syndrome patients
G. Baldassarre1, C. Rossi2, E. Banaudi1, M. Tartaglia3, A. Marinosci1, M. Silengo1, G. B. Ferrero1; 1 Department of Pediatrics, Torino, Italy, 2Laboratory of Medical Genetics Policlinco S. Orsola Maplighi, Bologna, Italy, 3Department of Cell Biology and Neurosciences, Istituto Superiore di Sanit, Roma, Italy.

Prader-Willi syndrome (PWS) is a genomic disorder mostly caused by 15q11-q13 deletions (70%) due to meiotic Non-allelic homologous recombination (NAHR) between flanking Low Copy Repeats (LCR). A higher rate of 15q11-q13-deletions and duplications was previously reported in spermatozoa of fathers with PWS affected offspring. The aim of this work was to analyze the frequency of deletions and duplications of other regions with similar features (7q11.23 and 22q11.2) in order to assess whether a higher risk of transmitting other genomic disorders is present in this subjects. Semen samples from 16 PWS fathers and 10 control donors were processed and analyzed by triple-color FISH as standardized in our laboratory. A customized combination of probes (Abbot Molecular Inc.) was used to assess the target regions allowing the discrimination between normal, deleted and duplicated sperm genotypes. A minimum of 10000 sperm were analized per sample and region. As a whole, higher rates of deletions and duplications were observed for 15q11-q13 (0.9%0.14) and 22q11.23 region (0.44%0.09) compared with the control population (0.46%0.07 and 0.27%0.05 respectively) (Mann-Whitney test; P<0.05). Interestingly, all individuals with significant increases of 7q11.23 and 22q11.2 had also been reported to have increases of 15q11q13-deletions and duplications. Moreover, a significant correlation were observed between the frequencies of deletions and duplications of 15q11-q13 and 7q11.23 regions (r=0.87; P=0.02) and between 15q11-q13 and 22q11.2 regions (r=0.89; P=0.002). Results suggest that other factors apart from architectural features in these regions could participate in the increases of NAHR events. Acknowledgements: Grant UAB.PIF/2007. c06.5* Genome-wide single cell array analysis for preimplantation genetic diagnosis of a complex chromosomal rearrangement carrier
E. Vanneste1,2, C. Melotte1, U. Ullman3, C. Staessen3, I. Liebaers3, T. Voet1, S. Debrock2, A. Pexsters2, C. Tomassetti2, J. Fryns1, T. DHooghe2, J. R. Vermeesch1; 1 Center for Human Genetics / K.U.Leuven - UZ Gasthuisberg, Leuven, Belgium, 2Leuven University Fertility Center / UZ Gasthuisberg, Leuven, Belgium, 3Center for Medical Genetics / UZ Brussels, Brussels, Belgium.

Noonan syndrome (NS) is a frequent autosomal dominant disorder caused by mutations in the genes of the RAS/MAPK pathway, being the most frequent PTPN11 and SOS1. Besides the well known post-natal heterogeneous clinical presentation characterized by short stature, congenital heart defects, facial and skeletal dysmorphisms, cryptorchidism, lymphatic dysplasia; the prenatal phenotype is not precisely defined, as well as the related prognostic factors. Several NS anomalies can be observed in fetuses, such as those ones related to lympathic dysplasia (increased nuchal translucency, cystic hygromas, hydrops fetalis, multiple effusions and polyhydramnios), cardiac defects, and growth retardation. In this retrospective study we evaluated the incidence of abnormal prenatal findings in a cohort of 40 NS patients including 28 PTPN11, 8 SOS1, 1 KRAS, 1 BRAF, 1 RAF1 and 1 SHOC2 mutated patients. Prenatal anomalies were found in 42.5% (17/40) of them : 40% (16/40) polydramnios, 17.5 % (7/40) increased fetal nuchal translucency, 17.5 % (7/40) hydrothorax and multiple effusions, 12.5% (5/40) premature rupture of membranes, 2,5 % (1/40) cardiac defects, 9 (22.5%) presenting multiple defects. In conclusion, prenatal anomalies can be found in an elevated fraction of NS patients, being the most common polyhydramnios, with a consistent portion of them presenting multiple anomalies. Interestingly, the observed anomalies do not correlate with the severity of the postnatal phenotype, both at birth and in the first years of life, suggesting that prenatal diagnosis of NS is not burdened by an adverse prognosis. C07.1 Identification of biomarkers for acute lymphoblastic leukemia (ALL) by allele-specific gene expression and DNA methylation analysis of primary ALL cells
L. Milani1, A. Lundmark1, J. Nordlund1, A. Kiialainen1, K. L. Gunderson2, T. Flaegstad3, E. Forestier4, M. Heyman5, G. Jonmundsson6, J. Kanerva7, K. Schmiegelov8, S. Sderhll9, M. G. Gustafsson10, G. Lnnerholm11, A. C. Syvnen1; 1 Molecular Medicine, Department of Medical Sciences, Uppsala University, Uppsala, Sweden, 2Illumina Inc., San Diego, CA, United States, 3Nordic Society of Pediatric Oncology, Norway, 4Department of Clinical Sciences, University of Ume,, Ume, Sweden, 5Karolinska University Hospital, Stockholm, Sweden, 6 Nordic Society of Pediatric Oncology, Reykjavik, Iceland, 7Nordic Society of Pediatric Oncology, Helsinki, Finland, 8Nordic Society of Pediatric Oncology, Copenhagen, Denmark, 9Nordic Society of Pediatric Oncology, Stockholm, Sweden, 10Clinical Pharmacology and Informatics, Department of Medical Sciences, Uppsala University, Uppsala, Sweden, 11Department of Womens and Childrens Health Medical Sciences, University Childrens Hospital,, Uppsala, Sweden.

Patients carrying a complex chromosomal rearrangement (CCR) have an increased risk for chromosomally unbalanced conceptions, which either do not implant, are spontaneously lost during gestation or lead to severely handicapped children. Preimplantation genetic diagnosis (PGD) can select against these embryos carrying unbalanced rearrangements, however only 7 loci can be screened and an indicationspecific preparation is needed using the standard FISH-technique. Microarray technology, on the contrary, can detect imbalances as small as 10 Mb genome-wide at the single cell level. We performed for the first time microarray-based PGD for a CCR-carrier with karyotype 46,XY,ins(3;2)(p23; q23q14.2),t(6;14)(p12.2; q13). In concurrence with the ethical committee, only the copy number status of the chromosomes involved in the CCR were diagnosed. Two PGD cycles with single embryo transfer were performed, which resulted in a clinical pregnancy. The embryo that gave rise to the pregnancy was normal (or balanced) for the inherited CCR, but carried however a trisomy 8 and nullisomy 9 in one of the two biopsied blastomeres. After 8 weeks of pregnancy the couple miscarried. Genetic analysis following embryo-hysteroscopy showed a diploid(90%)/tetraploid(10%) mosaic chorion, while the gestational sac was empty. No aneuploidy 8 was detected in the chorion, while 8% of the cells carried a monosomy for chromosome 9. We demonstrate that microarray enables to screen against the transmission of the unbalanced meiotic products that can derive from (complex) chromosomal rearrangements. In addition, the findings demonstrate that the genomic constitution of the implanted embryo is not representing the chromosomal rearrangements detected in a single blastomere.

We used genome-wide allele-specific expression (ASE) patterns to identify genes with cis-acting factors that regulate gene expression in acute lymphoblastic leukemia (ALL) cells. We determined the allelic expression levels of 8,000 genes in bone marrow or peripheral blood samples from 197 children with ALL, by quantitative genotyping of single nucleotide polymorphisms (SNPs) in RNA. This analysis identified 391 genes that displayed ASE. Extended promoter regions of these genes were subjected to DNA methylation analysis in cells from 401 patients with childhood ALL. We found that CpG sites located outside CpG islands had higher methylation levels and larger variability in CpG site methylation than CpG sites within CpG islands. We identified 47 genes with an inverse correlation between CpG site methylation and ASE in our ALL sample set, and for 24 genes we observed a correlation between CpG site methylation and ASE levels in individual ALL samples. Using supervised learning, we constructed multivariate classifiers by external cross-validation procedures, and identified genes where the DNA methylation levels allowed accurate discrimination between ALL cells and control cells, between T-lineage and B-cell precursor (BCP) ALL and between the main cytogenetic subtypes of BCP ALL. We also identified 20 genes with DNA methylation levels that predicted relapse of ALL. Most of the genes high-lighted in our study are novel, with no reported connection with ALL. Our findings open up perspectives for more detailed studies on DNA methylation as a

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molecular event that leads to ALL and affects the treatment response and clinical outcome. c07.2* correlation of telomere length shortening with promoter methylation profile of p16/Rb and p53/p21 pathways in breast cancer
Z. Barekati, R. Radpour, C. Kohler, R. Asadollahi, X. Zhong; Laboratory for Prenatal Medicine and Gynecologic Oncology, Basel, Switzerland. Wrzburg, Germany, 4Interdisciplinary Centre for Clinical Research, Universitts-Frauenklinik Wrzburg, Wrzburg, Germany.

Unregulated cell growth, a major hallmark of cancer, is coupled with telomere shortening. Measurement of telomere length could provide important information on cell replication and proliferation state in cancer tissues. Telomere shortening and its potential correlation with downregulation of cell-cycle regulatory elements were studied by the examination of relative telomere length and methylation status of the TP53, P21 and P16 promoters in tissues from breast cancer patients. Telomere length was measured in 104 samples (52 tumors and paired adjacent normal breast tissues) by quantitative PCR. Methylation profile of selected genes was analyzed in all samples using a matrixassisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Our results demonstrated a significant shortening of tumor telomere regions compared to paired adjacent normal tissues (P<0.001). Similarly, telomere lengths were significantly shorter in advanced stage cases and in those with higher histological grades (P<0.05). Telomere shortening in cancer tissues was correlated with a different level of hypermethylation in the TP53, P21 and P16 promoters (r=-0.33, P=0.001; r=-0.70, P<0.0001 and r=-0.71, P<0.0001, respectively). The results suggested that inactivation of p16/Rb and/or p53/ p21 pathways by hypermethylation may be linked to critical telomere shortening, leading to genome instability and ultimately to malignant transformation. Thus, telomere shortening and promoter hypermethylation of related genes both might serve as breast cancer biomarkers. c07.3* Discovery of molecular targets for therapy by expression profiling of thyroid cancer cases
D. Nikolova1,2, I. Dimova1, H. Zembutsu2, T. Sechanov3, K. Vidinov3, L. Kee4, R. Ivanova3, E. Becheva1, D. Toncheva1, Y. Nakamura2; 1 Department of Medical Genetics, Medical University Sofia, Sofia, Bulgaria, 2 Institute of Medical Science, University of Tokyo, Tokyo, Japan, 3Clinic of Endocrine Surgery, Medical University Sofia, Sofia, Bulgaria, 4University of Tokyo, Tokyo, Japan.

Screening is an unsolved problem for ovarian cancer, especially as late detection is equivalent to poor prognosis. Thus, we investigated whether ovarian cancer patients display characteristically deregulated miRNAs in blood cells that could form the basis of a test for disease progression or even for preventive screening. To this aim, blood-borne miRNA profiles from 15 patients with ovarian cancer and 15 age- and sex-matched healthy controls were compared and biostatistically evaluated. This showed that 51 out of >900 tested miRNAs were deregulated by unadjusted Students t-test. While some candidates had already been linked with cancer (e.g. miR-155), most had never been connected to a specific disease before. Bioinformatics further enabled us to define a miRNA profile which allowed for discrimination between blood samples of ovarian cancer patients and healthy controls with an accuracy and specificity of >70%. When only cancers of the serous subtype were considered and compared with an extended control group (n=37), sensitivity exceeded 90%. Taken together, our proof-of-principle study strengthens the hypothesis that neoplastic diseases generate characteristic miRNA fingerprints in blood cells. Thus, we propose that the combination of microarray-based miRNA-profiling from peripheral blood with other markers might improve the notoriously difficult but important screening for ovarian cancer. C07.5* Methylation Profiles of 22 Candidate Genes in Breast cancer Using High-throughput mALDi-tOF mass Array

R. Radpour, Z. Barekati, C. Kohler, X. Zhong; Laboratory for Prenatal Medicine and Gynecologic Oncology, Womens Hospital/Department of Biomedicine, University of Basel, Basel, Switzerland.

Thyroid cancer represents a heterogeneous entity and its clinical characteristics vary widely. Gene-expression profiling studies have identified a variety of potential molecular markers to help distinguish benign from malignant thyroid neoplasms. To elucidate the mechanisms underlying the progression and to identify novel therapeutic targets, we assessed the genome-wide expression in normal and tumor thyroid tissues. Pure populations of malignant and healthy cells were isolated by applying the technology of LCM. We compared the gene expression patterns of 18 malignant thyroid samples and 13 samples of normal epithelial non-transformed cells using the GeneChip trademark from Affymetrix. We identified 243 probes that were significantly differentially over-expressed and whose ratio tumor/control was more than 5 in more than 50% of the analyzed samples. Among the top overexpressed genes in thyroid carcinoma were: FN, CHI3L1, FAM20A, SEACAM6, NMU, LPL etc. Unsupervised hierarchical clustering separated papillary from medullar carcinoma samples. The differential expression of 7 genes was validated by quantitative real-time PCR. Two of those candidates (RGS4 and QPCT) were confirmed by immunohistochemistry, Northern blot analyses and further functional analysis, including siRNA experiments.Based on this result, we consider RGS4 as a good molecular target for therapy of thyroid cancer. Thus, the molecular signatures unique to thyroid carcinoma provide a molecular basis for therapeutic target discovery. c07.4* A new approach for ovarian cancer screening characterization of miRNA profiles in peripheral blood

Alterations of DNA methylation patterns have been suggested as biomarkers for diagnostics and therapy of cancers. Every novel discovery in the epigenetic landscape and every development of an improved approach for accurate analysis of the events may offer new opportunity for the management of patients. Using a novel high-throughput mass spectrometry on MALDI-TOF silico-chips, we determined quantitative methylation changes of 22 candidate genes in breast cancer tissues. For the first time we analysed the methylation status of a total of 42,528 CpG dinucleotides on 22 genes in 96 different paraffin-embedded tissues (48 breast cancerous tissues and 48 paired normal tissues). A two-way hierarchical cluster analysis was used to classify methylation profiles. In this study, 10 hypermethylated genes (APC, BIN1, BMP6, BRCA1, CST6, ESRb, GSTP1, P16, P21 and TIMP3) were identified to distinguish between cancerous and normal tissues according to the extent of methylation. Individual assessment of the methylation status for each CpG dinucleotide indicated that cytosine hypermethylation in the cancerous tissue samples was mostly located near the consensus sequences of the transcription factor binding sites. These hypermethylated genes may serve as biomarkers for clinical molecular diagnosis and targeted treatments of patients with breast cancer. c07.6 Unraveling the complexity of Primary and metastatic Ewings sarcoma Using Helicos single molecule sequencing
P. Kapranov1, R. J. Arceci2, J. Buckley3, G. Reaman4, P. Reynolds5, P. Sorensen6, J. Thompson1, P. Milos1, T. Triche3; 1 Helicos BioSciences Corporation, Cambridge, MA, United States, 2John Hopkins University, Baltimore, MD, United States, 3University of Southern California, Los Angeles, CA, United States, 4Childrens Oncology Group, Bethesda, MD, United States, 5Texas Tech University, Lubbock, TX, United States, 6University of British Columbia, Vancouver, BC, Canada.

S. F. M. Husler1, A. Keller2, A. Chandran3, J. Wischhusen4; 1 Universitts-Frauenklinik Wrzburg, Wrzburg, Germany, 2Biomarker Discovery Center Heidelberg and febit biomed GmbH, Heidelberg, Germany, 3 Graduate School for Life Sciences, Universitts-Frauenklinik Wrzburg,

Ewing sarcoma is a high-risk childhood cancer that represents a major treatment challenge, as survival has not improved significantly despite aggressive chemotherapy. Elucidation of molecular changes in progression of this cancer towards the metastatic and chemo-resistant states offers the potential to better understand the fundamentals of biological processes responsible for poor disease outcome. We have applied Helicos single-molecule sequencing to build unbiased wholegenome profiles of the transcriptome, methylome and DNA of two cell lines derived from the primary tumor of a Ewing sarcoma patient and her chemo-resistant metastasis. This simple system is ideal for generation of hypotheses on relationships between coding and non-coding RNAs, the epigenome, primary DNA sequence, and the malignant phenotype. Unlike work on primary tumors, it also allows for biologic validation.

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Large differences between the primary and metastatic cells exist at both the genomic and transcriptome levels, including differential expression and methylation as well as copy number and sequence variants suggesting clonal evolution from the primary to the metastatic tumor. A significant fraction of the genome was found to represent RNAs expressed at different levels between the cell lines, of which ~60% corresponded to un-annotated transcripts. In some instances, the latter could span large (100s of kb) genomic regions that contain no annotated transcripts. About 25% of the transcripts upregulated in the metastatic clone were found to be associated with promoter demethylation when compared to the primary tumor, suggesting that promoter demethylation is specifically associated with up-regulation of transcripts in the metastatic/chemo-resistant tumor in a selective manner. C08.1 Treacher Collins syndrome - detailed genetic and phenotypic analysis

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Fmn1 play an important role in mouse and chick limb development. The mouse limb deformity (ld) phenotype with digit reduction, syndactyly, radio-ulnar synostosis, variable renal defects and absent fibulae is caused by loss of Grem1 function. This could be due to either coding Grem1 homozygous mutations or homozygous deletions of the neighbouring Fmn1 gene, which also removes limb specific regulatory sequences of Grem1. Recent studies reinforce the hypothesis that a loss of Fmn1 protein could also contribute to the observed ld anomalies. In addition, an over-expression of Grem1 in developing chick limbs represses the programmed cell death in the interdigital mesenchyme, resulting in interdigital webbing and truncation of distal cartilage elements. We report here, for the first time, chromosomal imbalances in the GREM1-FMN1 region in individuals with limb defects. A 263Kb homozygous deletion of FMN1 was associated with oligosyndactyly, radioulnar synostosis, hearing loss and renal defects, features identical to ld mice. A 1.7Mb duplication encompassing both the GREM1 and FMN1 genes was detected in an isolated Cenani-Lenz-like oligosyndactyly of the hands, resembling the transgenic chick wings in which Grem1 was over-expressed. The phenotypes of the patients represent new entities/syndromes within the Cenani-Lenz clinical spectrum- (1) an autosomal recessive Oligosyndactyly, Radio-Ulnar Synostosis, Hearing loss and Renal defect syndrome and (2) an autosomal dominant Cenani-Lenz-like non-syndromic Oligosyndactyly. C08.3 An inherited arthritis is caused by mutations in TRPV4

D. Wieczorek1, S. Seland1, H. J. Ldecke1, S. Bhringer1,2, L. Klein-Hitpass3, C. Daumer-Haas4, N. Elcioglu5, H. Hameister6, H. Kayserili7, A. Kobelt8, D. Mller8, O. Rittinger9, S. Spranger10, G. Strobl-Wildemann11, J. Vodopiutz12, R. Brekelmans13, D. R. Lohmann1, U. Hehr14; 1 Institut fr Humangenetik, Universittsklinikum Essen, Essen, Germany, 2 Department of Medical Statistics and Bioinformatics, Leiden University Medical Centre, Leiden, Netherlands, 3Institut fr Zellbiologie (Tumorforschung), BioChip-Labor, Essen, Germany, 4Prnatal-Medizin Mnchen, Mnchen, Germany, 5Department of Pediatric Genetics, Marmara University Hospital, Istanbul, Turkey, 6genetikum, Neu-Ulm, Germany, 7Department of Medical Genetics, Istanbul Medical Faculty, Istanbul University, Istanbul, Turkey, 8Institut fr Medizinische Genetik, Klinikum Chemnitz gGmbH, Chemnitz, Germany, 9 Department of Pediatrics, Paracelsus Medical University Salzburg, Salzburg, Austria, 10Praxis fr Humangenetik, Klinikum Bremen-Mitte, Bremen, Germany, 11 Praxis fr Humangenetik, Ingolstadt, Germany, 12Department of Pediatrics and Adolescent Medicine, Medical University Vienna, Vienna, Austria, 13MRC Holland, Amsterdam, Netherlands, 14Zentrum fr Humangenetik, Universitt Regensburg, Regensburg, Germany.

The Treacher Collins syndrome (TCS, OMIM #154500) is an autosomal dominant condition characterised by craniofacial dysmorphism consisting of downslanting palpebral fissures, lower eyelid coloboma, hypoplasia of facial bones and microtia. Mutation analysis of the TCOF1 gene identifies pathogenic mutations in more than 90% of patients with typical TCS. In 226 patients referred to us as having TCS, we performed TCOF1 sequence and MLPA (26/28 exons) analyses. Point mutations scattered throughout the gene were identified in 78 of them, but no deletion or duplication was found in the remaining 148. By careful evaluation of all available clinical data of these 148 patients, we could confirm the suspected diagnosis of TCS in only 10 of them. These were then subjected to SNP-array analyses, but no deletion or duplication was found. We will report in detail on the clinical findings in 49 index patients, including 21 new ones, and their affected family members with a pathogenic mutation in the TCOF1 gene and will show average 2D faces we created from standardized photographs. We observed a wide interand intrafamilial phenotypic variability. Possibly stochastic variation or unidentified genetic modifiers, most likely not in cis might contribute to the phenotypic variability. Some members of families with molecularly proven TCS are so mildly affected that they do not even fulfill the minimal diagnostic criteria defined earlier (Teber et al., 2004). Thus, a genotype phenotype correlation can not be made. The widened phenotypic spectrum must be considered in genetic counselling. C08.2 Genomic rearrangements of the GREM1-FMN1 locus cause Oligosyndactyly, Radio-Ulnar synostosis, Hearing loss, Renal defects syndrome and cenani-Lenz-like non-syndromic Oligosyndactyly

R. Savarirayan1, S. Lamande1, Y. Yuan2, I. Gresshoff1, C. Little3, L. Rowley1, K. Kaluarachchi1, E. Botzenhart4, D. Amor1, W. Cole5, P. McIntrye6, J. Bateman1; 1 Murdoch Childrens Research Institute, Melbourne, Australia, 2Dept. Pharmacology, University of Melbourne, Melbourne, Australia, 33Raymond Purves Bone & Joint Research Laboratories, Sydney, Australia, 4 4Universitatsklinikum Freiburn, Institut fur Humangenetik und Anthropologie, Freiburg, Germany, 56Stollery Childrens Hospital/University of Alberta Hospital, Edmonton, AB, Canada, 6Department of Pharmacology, University of Melbourne, Australia, Melbourne, Australia.

Familial Digital Arthropathy-Brachydactyly (FDAB; OMIM 606835) is a dominantly inherited condition involving aggressive osteoarthritis of the fingers and toes and consequent shortening of the middle and distal phalanges. We show, in two unrelated families, that the disorder is caused by missense substitutions (G270V, R271P and F273L) in the intracellular ankyrin repeat domain of the transient receptor potential cation channel TRPV4. Functional testing of mutant TRPV4 in stably transfected HEK293 cells revealed the mutant proteins showed poor cell surface localization, despite being expressed at similar levels to wild type protein, and calcium influx in response to the TRPV4 agonist GSK1016790A was significantly reduced. Others have recently shown that gain of function TRPV4 mutations cause a range of skeletal dysplasias and peripheral neuropathies. Our data, showing tightly clustered TRPV4 mutations that reduce channel activity in a third phenotype, inherited osteoarthritis, demonstrate the importance of TRPV4 activity in articular cartilage homeostasis and raises the possibility that this cation channel might play a role in age-related osteoarthritis. C08.4* Mutations in PITX1 cause a human patellar malformation syndrome: Delineation of a recognisable lower-limb phenotype

E. M. H. F. Bongers1, J. Schoots1, F. Stock2, N. V. A. M. Knoers1, J. W. M. Gardeniers3, A. van Kampen3, J. de Rooij4, L. Hoefsloot1; 1 Dept of Human Genetics, Radboud University Nijmegen Medical Centre, Nijmegen, Netherlands, 2Institut fr Humangenetik, Universittsklinikum Leipzig AR, Leipzig, Germany, 3Dept of Orthopaedic Surgery, Radboud University Nijmegen Medical Centre, Nijmegen, Netherlands, 4Dept of Radiololy, Radboud University Nijmegen Medical Centre, Nijmegen, Netherlands.

B. I. Dimitrov1, T. Voet1, L. De Smet2, J. R. Vermeesch1, K. Devriendt1, J. Fryns1, P. Debeer1,2; 1 Centre for Human Genetics, University Hospitals Leuven, Leuven, Belgium, 2 Department of Musculoskeletal Science, Division of Orthopaedics, University Hospitals Leuven, Pellenberg, Belgium.

Limb development is a complex process requiring proper spatio-temporal expression of a network of limb specific morphogens. Grem1 and

The identification of gene defects in human patellar malformation syndromes and mutant animal models proved to be important in unravelling key molecules involved in patellar development and musculoskeletal patterning. The nail patella syndrome phenotype emphasised that the LIM homeodomain transcription factor LMX1B is critical in dorsoventral and anteroposterior patterning of the upper- and lowerlimbs, as well as the formation of patellar/iliac/radial bones. Phenotype studies of the small patella syndrome (SPS) determined that the T-box transcription factor TBX4 is implicated in dorsoventral and anteroposterior patterning of the lower-limb and late skeletal development of patellar/ischiopubic/(meta)tarsal bones. We aimed at elucidating the

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molecular cause in four classical SPS families and six families with a SPS-like phenotype, in which sequencing of TBX4,TBX2, and LMX1B was found normal. The paired-like homeodomain transcription factor PITX1 was selected as a strong candidate gene for congenital patellar, pelvic and foot malformations, based on its role as an upregulator of Tbx4 in the determination of lower-limb identity in chicken and mice. Pitx1 knockout mice showed absent patellae and lower-limb bones resembling an upper limb-like morphology. We identified two different putative loss-of-function mutations (one missense and one nonsense) in PITX1 in one four-generation and one two-generation SPS-like family, respectively. The phenotypic spectrum comprised aberrant patellar size and morphology, unilateral clubfoot and iliac/tibial/fibular/talus/calcaneal bone malformations. The present identification of PITX1 mutations in man, together with the phenotype of animals lacking Pitx1 delineates a recognisable autosomal dominant patellar malformation syndrome resulting from disrupted anteroposterior patterning and leftright directional asymmetry of the lower-limb. C08.5* TRPV4-related skeletal dysplasias: a clinical, radiographic, and molecular study in 18 families.

0
tients, aged eight months to 38 years) with FOP and on molecular findings of 13 of them. Most of them show a typical hypoplasia/aplasia and valgus deviation of the great toe (17/20). A common symptom are painful swellings on shoulders and face beginning to arise approximately at an age of three years (15/20). Hand anomalies, such as thumb hypoplasia (18/20) and clinodactyly of the fifth finger (15/20) are common. Some patients have hypoacusis. Further features are restricted mobility of the spine and contractures of other joints as a result of heterotopic ossifications. Some patients show craniofacial features like hypomimia, sparse eyebrows and hair and teeth anomalies. Sequence analysis of the ACVR1 gene were performed on 13/20 patients. Ten patients show the typical mutation p.R206H. Two patients with an atypical phenotype show different mutations (G328W and G356D). We analysed the metacarpophalangeal profile (MCPP) on X-rays of seven patients. We suggest that there is a typical MCPP pattern in classical FOP, which could be used as an additional diagnostic tool. We discuss the clinical, radiological and molecular findings of our patients and compare them with the literature. Our study contributes to the understanding of the FOP phenotype and possible genotype-phenotype correlations. c09.1 chromosome-wide mapping of long-range interactions involved in smith-magenis and Potocki-Lupski syndromes

E. Andreucci1,2, J. Bateman3, S. Aftimos4, E. Haan5, W. Hunter6, B. Kerr7, G. McGillivray2, R. J. McKinlay Gardner4, M. Patricelli8, D. Sillence9, S. Lamand3, R. Savarirayan2; 1 Dept. of Clinical Pathophysiology and Medical Genetics Unit, Meyer Childrens Hospital, University of Florence, Florence, Italy, 2Genetic Health Services Victoria and Murdoch Childrens Research Institute, Royal Childrens Hospital, Parkville, Victoria 3052, Australia, 3Murdoch Childrens Research Institute, Royal Childrens Hospital, Parkville, Victoria 3052, Australia, 4Northern Regional Genetics Service, Auckland City Hospital, Auckland, New Zeland, 5South Australian Clinical Genetics Services, SA Pathology, Womens and Childrens Hospital and Department of Paediatrics, University of Adelaide, Adelaide, Asutralia, 6Paediatrician Midcentral District Heath Board Palmerston North New Zealand, 7Genetic Medicine, St Marys Hospital, Manchester M139WL, UK, 8Servizio di Genetica, Ospedale San Raffaele, Milan, Italy, 9Department of Genetic Medicine, University of Sydney, New South Wales, Australia.

N. Gheldof1, B. Lajoie2, G. Ricard1, J. Chrast1, J. Molina3, J. R. Lupski4, K. Walz3, J. Dekker2, A. Reymond1; 1 University of Lausanne, Lausanne, Switzerland, 2University of Massachusetts Medical School, Worcester, MA, United States, 3University of Miami, Miami, FL, United States, 4Baylor College of Medicine, Houston, TX, United States.

The transient receptor potential vanilloid 4 protein (TRPV4) is a calcium-permeable ion channel that responds to many different stimuli, is widely expressed, and participates in an extraordinarily wide range of physiologic processes. Autosomal dominant brachyolmia, spondylometaphyseal dysplasia Kozlowski type (SMDK) and metatropic dysplasia are three distinct skeletal dysplasias which share some common features, including short stature, platyspondyly, and progressive scoliosis. SMDK and metatropic dysplasia also have significant but variable metaphyseal involvement on radiographs. In the past two years, mutations in the gene encoding TRPV4 have been found to be responsible for these three skeletal phenotypes, confirming that they are allelic disorders and suggesting that they might represent different parts of a phenotypic spectrum. We analysed the clinical, radiographic and molecular data on 21 patients from 18 families, all of whom had a clinical diagnosis of one of the conditions described above: 12 with metatropic dysplasia; 3 with SMKD; and 3 with brachyolmia. Our study identified 7 different mutations in 14 out of the 18 studied families, two previously described, and 5 novel. These data have uncovered new genotype-phenotype correlations and suggest that these three conditions represent a continuum of severity with areas of phenotypic overlap between conditions, even within the same family. We hope that these data will also add to the understanding of the molecular basis of these disorders and identify possible pharmacologic targets for therapy. C08.6 Clinical and molecular findings on 20 patients with fibrodysplasia ossificans progressiva

Copy number variations (CNVs) affect expression levels of the genes that map within the affected region, but also of genes located in the flanking regions. To understand the mechanisms at play, we studied gene expression and chromatin conformation in mice models of SmithMagenis (SMS) and Potocki-Lupski (PTLS) syndromes, containing a microdeletion and its reciprocal microduplication, respectively, on mouse chromosome 11 (MMU11). We profiled the transcriptome of embryonic fibroblasts of mice with one, two, three and uniallelic two copies of the SMS/PTLS region in an otherwise identical genetic background. As expected, the most differentially expressed transcripts are mapping to the SMS/PTLS interval, but a significant proportion of most differentially expressed genes also map to the rest of MMU11. We hypothesized that these chromosome-wide effects might be caused by changes in long-range interactions along the entire chromosome. We therefore analyzed the chromatin structure of these four mouse strains by using the chromosome conformation capture carbon copy (5C) technology. This will allow comparing the chromatin structure and the presence of physical contacts between functionally interacting genomic elements in genotypes differing only by the number of copies of a CNV, and correlating these interaction maps with the observed differential gene expression. Detailed investigations of the different mechanisms by which CNVs alter the architecture of chromosomes are warranted to shed light on how CNVs influence gene expression. In addition, this study will provide a first comprehensive chromatin interaction map of an entire mouse chromosome. C09.2* Kidney-specific inactivation of Ofd1 leads to renal cystic disease associated to upregulation of the mtOR pathway
B. Franco1, D. Iaconis1, A. Barra1, A. Cantone2, N. Messaddeq3, G. Capasso2, P. Dolle3, P. Igarashi4, A. Zullo1; 1 Telethon Institute of Genetics and Medicine-TIGEM, Naples, Italy, 2Chair of Nephrology, Second University of Naples, Naples, Italy, 3Institut de Gntique et de Biologie Molculaire et Cellulaire, Illkirch Cedex, France, 4Department of Internal Medicine and Division of Basic Sciences, University of Texas Southwestern Medical Center, Dallas, TX, United States.

I. Stefanova1, C. Grnberg2, G. Gillessen-Kaesbach1; 1 Institut fr Humangenetik, Lbeck, Germany, 2Klinik fr Orthopdie und Unfallchirurgie, Allg. Krankenhaus Hagen, Hagen, Germany.

Fibrodysplasia ossificans progressiva (FOP) (MIM 135100) is a rare genetic disorder of progressive extraskeletal ossifications. The heterozygous mutation c.617G>A (p.R206H) in the ACVR1 gene is regarded as the cause of FOP in classically affected individuals worldwide (Shore et al.2006). We report on clinical findings of 20 patients (10 female, 10 male pa-

The Oral-Facial-Digital type I syndrome (OFDI; MIM 311200) is a rare syndromic form of inherited renal cystic disease. It is transmitted as an X-linked dominant, male lethal disorder and is caused by mutations in the OFD1 gene. Previous studies demonstrated that OFDI belongs to the growing number of disorders ascribed to dysfunction of primary cilia. We generated a conditional inactivation of the mouse Ofd1 gene using the Ksp-cre transgenic line, which resulted in a viable model characterized by renal cystic disease and progressive impairment of renal function. The study of this model allowed us to demonstrate that

Concurrent Sessions
primary cilia initially form and then disappear after the development of cysts, suggesting that the dysfunction of primary cilia is a consequence rather than the primary cause of renal cystic disease. Immunofluorescence and western blotting analysis revealed upregulation of the mTOR pathway in both dilated and non dilated renal structures. Treatment with rapamycin, a specific inhibitor of the mTOR pathway, resulted in a significant reduction in the number and size of renal cysts and a decrease in the cystic index compared with untreated mutant animals, suggesting that dysregulation of this pathway in our model is mTOR-dependent. The animal model we have generated could thus represent a valuable tool to understand the molecular link between mTOR and cyst development, and eventually to the identification of novel drug targets for renal cystic disease. c09.3 Lack of mid1, the mouse ortholog of the Opitz syndrome gene, causes abnormal development of the anterior cerebellar vermis
A. Lancioni1, M. Pizzo1, B. Fontanella1, R. Ferrentino1, L. M. R. Napolitano2, E. De Leonibus1, G. Meroni2,1; 1 Telethon Institute of Genetics and Medicine, Naples, Italy, 2Cluster in Biomedicine (CBM), Trieste, Italy.

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therapy may be an important option for parents faced with the prenatal diagnosis of a genetically affected child. c09.5* the forkhead transcription factor FOXL2 is sumoylated in both human and mouse: sumoylation affects its stability, localization, and activity
M. Marongiu1, M. Deiana1, A. Meloni1, L. Marcia1, A. Puddu1,2, A. Cao1, D. Schlessinger3, L. Crisponi1; 1 Istituto di Neurogenetica e Neurofarmacologia, Consiglio Nazionale delle Ricerche, Monserrato (CA), Italy, 2Universit degli Studi di Cagliari, Cagliari, Italy, 3National Institute on Aging, NIH, Baltimore, MD, United States.

Opitz G/BBB Syndrome (OS) is a genetic disorder characterized by midline developmental defects. Male patients with the X-linked form of OS, caused by loss-of-function mutations in the MID1 gene, show high variability of the clinical signs. MID1 encodes a ubiquitin ligase that controls Phosphatase 2A but its role in the pathogenesis of the disease is still unclear. Here we report a mouse line carrying a non-functional ortholog of the human MID1 gene, Mid1. Mid1 null mice show the brain anatomical defect observed in patients, i.e. hypoplasia of the anterior portion of the medial cerebellum, the vermis. We found that the presence of this defect correlates with motor coordination, procedural and non-associative learning impairments. The defect is limited to the most anterior lobes of the vermis, the region of the developing cerebellum adjacent to the dorsal midbrain. Analyses at mid-gestation reveal that lack of Mid1 causes the shortening of the posterior dorsal midbrain; the rostralization of the midbrain/cerebellum boundary; and the downregulation of a key player in the development of this region, Fgf17. Thus, lack of Mid1 causes a mis-specification of the midbrain/cerebellar boundary that results in an abnormal development of the most anterior cerebellar lobes. This animal model provides a tool for further in vivo studies of the physiological and pathological role of the Mid1 gene and a system to investigate the development and function of anterior cerebellar domains. c09.4* Global gene transfer in the cNs and phenotypic correction of mLD model mice by systemic neonatal injection of serotype 9 AAV vector
N. Miyake, K. Miyake, N. Asakawa, M. Okabe, M. Yamamoto, T. Shimada; Nippon Medical School, Tokyo, Japan.

The FOXL2 forkhead transcription factor is expressed in ovarian granulosa cells, and when mutated causes the Blepharophimosis, Ptosis and Epicanthus Inversus Syndrome (BPES) and predisposes to premature ovarian failure. Inactivation of Foxl2 in mice demonstrated its indispensability for female gonadal sex determination and ovary development and revealed its antagonism of Sox9, the effector of male testis development. To define FOXL2 regulatory activities we looked for interacting proteins. Based on yeast two-hybrid screening, we found that FOXL2 interacts with PIAS1 and UBC9, both parts of the sumoylation machinery. We confirmed the interactions by co-immunoprecipitation and we demonstrated that human FOXL2 is sumoylated in transfected cell lines, and that endogenous mouse Foxl2 is comparably sumoylated. Confocal microscopy allowed us to demonstrate that FOXL2 co-localizes with SUMO-1 in structures resembling PML (promyelocytic leukaemia) bodies in transfected cells and that FOXL2, SUMO1 and UBC9 colocalize in vivo in 4 weeks old mouse ovary. We identified 7 putative sumoylation sytes using Abgent SUMOplotsoftware, and created FOXL2 mutants in which the lysines of the higher score putative sumoylation sytes (K25, K87, K114, K150) were changed to arginine. Our results indicate that K114 and K150 are involved in FOXL2 sumoylation and nuclear localization and that all mutations influenced FOXL2 transcriptional activity. Furthermore we demonstrated that sumoylation results in an increase of FOXL2 protein levels, likely due to an increase in protein stability. It is intriguing that similar sumoylation and regulatory consequences have also been reported for SOX9, the male counterpart of FOXL2 in somatic gonadal tissues. c09.6 sAHA ameliorates the smA phenotype in two mouse models for spinal muscular atrophy
M. Riessland1, B. Ackermann1, A. Frster1, M. Jakubik1, J. Hauke1, L. Garbes1, I. Fritzsche2, Y. Mende1, I. Blumcke2, E. Hahnen1, B. Wirth1; 1 Institute of Human Genetics, Institute of Genetics and Center for Molecular Medicine Cologne, Cologne, Germany, 2Department of Neuropathology, University of Erlangen, Erlangen, Germany.

Lysosomal storage diseases (LSDs) are important targets for enzyme replacement and gene therapy. The success of gene therapy for LSDs with neurological involvement such as metachromatic leukodystrophy (MLD) depends on the development of efficient delivery of lysosomal enzymes and/or vectors across the blood-brain barrier (BBB) to achieve wide distribution of enzyme throughout the brain. Since both the immune system and the BBB are developmentally immature during the perinatal period, neonatal gene transfer may be a highly promising strategy to treat genetic neurological disorders. To treat MLD mice, we generated serotype 9 AAV vector expressing human arylsulfatase A (AAV9/ASA) and IV injected into neonatal MLD mice (n=7). ELISA analysis showed that sustained expression of ASA was detected in the brain (cerebral cortex: 24.58.7, cerebellum: 7.84.5 ng/mg protein) for more than one year. Alcian blue staining and quantitative analysis of sulfatide contents by biochemical assay showed significant decrease of the amount of stored sulfatide in AAV9/ASA treated MLD mice compared to non-treated mice (cerebral cortex: 12.05.3 vs. 29.712.7, p<0.03; cerebellum: 34.86.3 vs. 73.25.0, mg/mg protein, p<0.05). Furthermore, in the behavior test, AAV9/ASA treated mice showed a significant improvement in their ability to traverse narrow balance beams, as compared to non-treated MLD mice (Latency: 9.21.5 vs. 13.00.4 sec, P<0.05; Slips: 3.51.9 vs. 4.31.4 times, P<0.05). These data clearly demonstrate that MLD model mice can be treated by systemic neonatal injection of AAV9/ASA. Therefore, neonatal gene

Proximal spinal muscular atrophy (SMA) is a common autosomal recessively inherited neuromuscular disorder determined by functional impairment of -motor neurons within the spinal cord. SMA is caused by functional loss of the survival motor neuron gene 1 (SMN1), whereas disease severity is mainly influenced by the number of SMN2 copies. SMN2, which produces only low levels of full-length mRNA/protein, can be modulated by small molecules and drugs, thus offering a unique possibility for SMA therapy. Here, we analysed suberoylanilide hydroxamic acid (SAHA), a FDAapproved histone deacetylase inhibitor, as potential drug in two severe SMA mouse models each carrying two SMN2 transgenes: US-SMA mice with one SMN2 per allele (Smn-/-;SMN2tg/tg) and Taiwanese-SMA mice with two SMN2 per allele (Smn-/-;SMN2tg/wt), both on pure FVB/N background. The US-SMA mice were embryonically lethal with heterozygous males showing significantly reduced fertility. SAHA-treatment of pregnant mothers rescued the embryonic lethality giving rise to SMA offspring. By using a novel breeding strategy for the Taiwanese model (Smn-/-;SMN2tg/tg x Smn-/+ mice) we obtained 50% SMA offspring that survive ~10 days and 50% control carriers in each litter. Treatment with 25 mg/kg/2x/day SAHA increased lifespan of SMA mice by 30%, significantly improved motor function abilities, reduced degeneration of motor neurons within the spinal cord and increased the size of neuromuscular junctions and muscle fibers compared to vehicle-treated SMA mice. SMN RNA and protein levels were significantly elevated in various tissues including spinal cord and muscle. Hence, SAHA, which lessens the progression of SMA, might be suitable for SMA therapy.

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c10.1 copy number variable regions in 13 European populations

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type VIII OI (OMIM #610915). We have identified a LEPRE1 mutation, IVS5+1G>T, in unrelated African Americans (AA) and contemporary West Africans (WA) immigrants. Screening of gDNA from a Mid-Atlantic AA cohort determined a carrier incidence of 0.30-0.50% for this mutation, predicting occurrence of homozygous lethal type VIII OI in about 1/250,000 births in this population. To trace the mutationorigin, we screened gDNA from a contemporary WA cohort. Nineteen of 1284 unrelated individuals (1.48%) from Nigeria and Ghana were heterozygous carriers, half of whom were from the Yoruba or Ibo ethnic groups of Nigeria. The high carrier frequency for this founder mutation among WAs predicts that this mutation alone would cause recessive OI in 1/18,250 births in WAs, equal to the incidence of de novo dominant OI in North America. To estimate the mutation age, we examined microsatellites and short tandem repeats spanning 4.2 MB surrounding the LEPRE1 gene. Disease allele haplotypes were determined from 12 contemporary WA and 3 AA families. WA carriers share a haplotype of 175-425Kb. Using the linkage disequilibrium analysis method of Rannala & Slatkin (2000), the mutation was estimated to have originated 800-960 years ago (1050-1210 C.E.). This timing is consistent with the model that this West African allele was transported to the Americas via the Atlantic slave trade (1450-1860 C.E.). c10.3 Paleogenetic study for reconstruction of genealogy of first Moldavian princes from 14th century
A. Rodewald1, G. Cardos2, C. Tesio3; 1 Institute of Human Biology, Hamburg, Germany, 2National Institute Victor Babes, Bucharest, Romania, 3Faculty of Biology- University Bucharest, Bucharest, Romania.

T. Esko1,2, G. Escramis3, R. Rabionet3, P. Palta2, M. Nelis1, F. Zimprich4, D. Toncheva5, M. Macek6, L. Peltonen7,8, B. Melegh9, D. Toniolo10, P. Gasparini11, J. Klovins12, V. Kuinskas13, J. Lubinski14, S. Limborska15, S. E. Antonarakis16, C. M. van Duijn17, M. Remm2, X. Estivill3, A. Metspalu1,2; 1 Estonian Genome Center, University of Tartu, Tartu, Estonia, 2Institute of Molecular and Cell Biology, University of Tartu, Tartu, Estonia, 3Center for Genomic Regulation (CRG-UPF) and CIBERESP, Barcelona, Spain, 4 Department of Clinical Neurology, Medical University of Vienna, Vienna, Austria, 5Department of Medical Genetics, Medical University of Sofia, Sofia, Bulgaria, 6Department of Biology and Medical Genetics, Cystic Fibrosis Centre, University Hospital Motol and 2nd School of Medicine, Charles University Prague, Prague, Czech Republic, 7Wellcome Trust Sanger Institute, Cambridge, United Kingdom, 8Institute of Molecular Medicine, Biomedicum, Helsinki, Finland, 9Department of Medical Genetics and Child Development, University of Pcs, Pcs, Hungary, 10Division of Genetics and Cell Biology, San Raffaele Research Institute, Milano, Italy, 11Medical Genetics, Department of Reproductive Sciences and Development, IRCCS-Burlo Garofolo, University of Trieste, Trieste, Italy, 12Latvian Biomedical Research and Study Center, Riga, Latvia, 13Department of Human and Medical Genetics, Vilnius University, Vilnius, Lithuania, 14International Hereditary Cancer Center, Pomeranian Medical University, Szczecin, Poland, 15Institute of Molecular Genetics, Russian Academy of Science, Moscow, Russian Federation, 16Department of Genetic Medicine and Development, University of Geneva Medical School, Geneva, Switzerland, 17Department of Epidemiology, Erasmus Medical Center, Rotterdam, Netherlands.

At present time there is no comprehensive copy number variable region (CNVR) map of European population although it is a prerequisite for the European-wide collaborative GWA studies. Most of the efforts so far have been directed into the characterization of the fine-scale CNVR structure in HapMap but Europe as a whole, especially the Eastern part, has not been covered in sufficient detail. We have studied about 2,000 individuals from thirteen cohorts from Europe (including about 900 Estonians) and focused on describing the genetic structure, especially the copy number variable regions (CNVR), in Eastern Europe and in Europe as whole. In total we identified more than 2,500 CNVR in 1,080 individuals (including only 100 randomly chosen Estonians). When analyzing only the Estonians, we identified more than 5,010 CNVR (maf cut-off 1%) and the majority of CNVRs had a very low frequency in the population. For the identified regions, principal component (PC) analysis were applied and the resulting map was not as informative geographically as in the case of SNP data. The CNVRs included in the 20 first PC show an important contribution from the HLA region, and the Immunoglobulin variable regions, while GO pathway analysis identifies a cluster of genes for sensory perception of chemical stimulus. In addition to SNPs, CNVs are being used as additional genetic markers in GWAS studies for disease gene mapping. The results presented here add new data to European genetic map of the structural variations that will greatly facilitate inter-population genetic studies. c10.2 A founder mutation in LEPRE1 causes lethal recessive type Viii Osteogenesis imperfecta and occurs in West Africans and African Americans
W. A. Cabral1, A. M. Barnes1, A. Adeyemo2, K. Cushing3, D. Chitayat4, F. D. Porter5, S. R. Panny6, F. Majid6, T. R. Rebbeck7, S. A. Tishkoff8, J. E. BaileyWilson9, L. C. Brody3, C. N. Rotimi2, J. C. Marini1; 1 Bone and Extracellular Matrix Branch, NICHD, NIH, Bethesda, MD, United States, 2Center for Research on Genomics and Global Health, NHGRI, NIH, Bethesda, MD, United States, 3Genome Technology Branch, NHGRI, NIH, Bethesda, MD, United States, 4Hospital for Sick Children, Mount Sinai Hospital, Toronto, ON, Canada, 5Heritable Disorders Branch, NICHD, NIH, Bethesda, MD, United States, 6Maryland Department of Health and Mental Hygiene, Baltimore, MD, United States, 7University of Pennsylvania School of Medicine and Abramson Cancer Center, Philadelphia, PA, United States, 8Departments of Genetics and Biology, University of Pennsylvania, Philadelphia, PA, United States, 9Inherited Disease Research Branch, NHGRI, NIH, Baltimore, MD, United States.

We performed paleomolecular genetic analysis on skeletal remains of 7 princes of Moldavia from the last half of the 14th century,buried in the Saint Nicholas Church from Radauti-Romania, in order to reconstruct their genealogy. We used the Amelogenin gene for identifying the genetic sex of the old individuals and we analysed mitochondrial (HVR I region)and nuclear DNA (VWA31A, TPOX, DYS392 and DYS393 Microsatellites)polymorphisms to reveal their genetic kinship along both maternal and paternal lineages.Ancient DNA was extracted by a phenol-chloroform-based method and amplified by PCR.The HVR I mitochondrial DNA sequences and nuclear DYS392 and DYS393 polymorphic markers were sequenced, and the other nuclear DNA markers were separated and identified on Polyacrylamide gels( Agstained).Our results revealed the genetic kinship between the 7 Moldavian princes, showing two male lines with a maternal linkage between the two groups. The first male line consisted of two individuals close related to each other and differed from the other 5 individuals also closed related among them as concerns the paternal line. This evidence argued that two princely dynasties succeeded on the Moldavian sceptre in the last half of the 14th century,contrary to the historiography version which had considered the existence of only one dynasty and succession exclusively along the paternal line. Acknowledgement:we thanks to:Dr. Nicolae Miritoiu and Lia and Adrian Batrana for providing skeletal remains;Angelika Kroll for technical support c10.4* modelling haplotype structure in isolate populations for population sequencing
K. Palin, K. Rehnstrom, L. Peltonen, R. Durbin; The Wellcome Trust Sanger Institute, Hinxton, Cambridge, United Kingdom.

Recessive osteogenesis imperfecta (OI) is caused by defects in cartilage-associated protein (CRTAP) or prolyl 3-hydroxylase 1 (P3H1/ LEPRE1), which form the collagen prolyl 3-hydroxylation complex with cyclophilin B (PPIB). Deficiency of P3H1 causes severe to lethal

Isolated founder populations are a powerful tool for disease genetics. By sequencing a modest number of individuals, we should be able to identify most founding haplotypes present in current individuals, then use long-range phasing and imputing to derive accurate and near complete genome sequence for all genome wide genotyped individuals within the population. To design such a study, we have developed a method to fit population history simulations to the observed pairwise IBS segment length distribution from dense genotype data. Using this approach with 7505 markers on chromosome 20, we created a genetic model for Kuusamo, a well known isolate population in North Eastern Finland. According to historical records, Kuusamo had small indigenous Lapp population until its resettlement by 34 families, totalling 136 chromosomes, in the 1680s. The current census population size is 25108, including some recent immigrants. Our best fit model has a population of 100 founding individuals ex-

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panding first rapidly, then more slowly, to a current effective population size of 7930 in 12 generations, rejecting significant historic immigration after the initial resettlement. The simulation suggests that out of the 200 founding haplotypes only 10810 survive today per locus, the rest having been lost by genetic drift. On average, 80% of a present day chromosome is inherited from the founders in fragments of length 6cM or more. The results suggest that a sample of 200 individuals would cover 93-98% of current indigenous chromosome population. We are exploring using this approach to model other European isolates. c10.5* inverse mapping approach implies the role of large cNVs in intellectual deficits and learning difficulties in a population cohort
O. P. H. Pietilinen1,2, K. Rehnstrm2, E. Jakkula1, S. Service3, E. Congdon3, C. Tilgmann2, T. Paunio1, S. Ripatti1, M. Jrvelin4,5, M. Isohanni6, C. Sabatti7, A. Palotie2, N. B. Freimer3, L. Peltonen2,1; 1 Institute for Molecular Medicine Finland FIMM, Helsinki, Finland, 2Wellcome Trust Sanger Institute, Hinxton, Cambridge, United Kingdom, 3Center for Neurobehavioral Genetics, Gonda Center, University of California Los Angeles, Los Angeles, CA, United States, 4Department of Epidemiology and Public Health, Imperial College London, London, United Kingdom, 5Department of Child and Adolescent Health, Institute for Health and Welfare, Oulu, Finland, 6 Department of Psychiatry, Institute of Clinical Medicine, University of Oulu, Oulu, Finland, 7Department of Human Genetics , University of California Los Angeles, Los Angeles, CA, United States.

kg/m2 difference). Gene expression and genotyping data was generated from 349 siblings using the Affymetrix Human U133 Plus 2.0 and Illumina Infinium Human 610K Quad platforms, respectively. RNA was extracted from subcutaneous adipose tissue and DNA was extracted from blood. Analysis was performed modelling genotyping signal intensity levels (LogR Ratio and B Allele Frequency) and gene expression levels in a variance component framework. Results: We detected more than 10 genomic regions where signal intensity was significantly associated with expression levels. For example, in a region on chromosome 6 overlapping the MHC, we identified 34 markers associated with transcript expression level at genome wide significance (p10-7). As expected, these regions are listed in the Database of Genomic Variants, i.e. they are CNVs. Conclusions: Using quantitative analysis of raw Illumina microarray data, we have identified more than 10 genomic regions that are associated with expression levels (p10-7). We will investigate these further, comparing them to the genes that are differentially expressed between obese and lean subjects, to identify candidate genes for obesity. c11.1 Policy Recommendations of the PPPc on direct-toconsumer genetic testing for health purposes

The availability of genome wide data on representative population samples provides an opportunity to apply a strategy of inverse mapping for correlating human traits with genotypes. Whereas the traditional forward mapping aims to define genotypic sharing accounting for a common phenotype among group of individuals, the inverse mapping seeks to discover phenotypic features shared among individuals demonstrating allelic similarity. This approach would avoid the inherent imprecision in phenotype definition that makes it difficult to determine a priori which individuals are sharing a phenotype. Here we provide a proof of principle of this inverse mapping approach by systematically scanning all CNVs >500 kb in a population cohort (N= 4,932). The participants were drawn from a prospective birth cohort of all individuals born in 1966 in North of Finland. Routine follow-ups have resulted in extensive phenotype database collected from the study participants. We identified 634 large CNVs observed in 529 individuals. To narrow down our data inquiries to a reasonable number of phenotypes, we focused on a category of traits postulated to relate with previous CNV findings in neuropsychiatric deficits. We observed significantly higher frequencies of cognitive defects defined as 8 among carriers of large deletions (5.0%) compared to non carriers (1.4%) (p <0.0024). Intriguingly, the deletion carriers were also more likely to present with sub clinical learning difficulties than the general population (10% vs. 3.9%; p=0.00088). The study highlights the opportunity to utilize inverse mapping as a strategy to characterize phenotypic consequences related to genetic variants in an unbiased population sample. c10.6* A novel approach to analysis of raw illumina microarray data to assess the contribution of copy number variations to obesity and gene expression

P. Borry1, C. Patch2, M. Cornel; on behalf of the PPPC3; 1 Centre for Biomedical Ethics and Law, 3000, Belgium, 2Guys Hospital, London, United Kingdom, 3VU Amsterdam, Amsterdam, Netherlands.

An increasing number of private companies are now offering direct-toconsumer (DTC) genetic testing services, ranging from tests for single gene, highly penetrant disorders to susceptibility testing for genetic variants associated with common complex diseases or with specific traits. The Professional and Public Policy Committee of the European Society of Human Genetics is concerned about the way commercial companies are currently introducing new genetic tests into the market and outside of the scope of the traditional health care system. Therefore, it is currently in the process of drafting policy recommendation on the topic of direct-to-consumer sales and/or advertising of genetic tests for health purposes. This presentation wants to draw the attention to this document and intensify the debate on this topic. The presentation will discuss following topics: the right to information, the advertising of genetic tests, the quality of genetic testing services (the quality of the genetic tests -in terms of validity and utility-, the quality of laboratories and the quality of the persons providing the genetic services), the individualized medical supervision, information and genetic counseling, informed consent, genetic testing of minors, respect for private life, research, oversight of genetic testing, and the impact on the healthcare system. c11.2* Reporting genetic research results: A quasi-experimental approach to understanding researchers judgments
R. Z. Hayeems, F. A. Miller, L. Li, J. P. Bytautas; University of Toronto, Toronto, ON, Canada.

J. C. Andersson1,2, A. J. Walley1, J. S. El-Sayed Moustafa1, P. Jacobson2, M. Jerns2, A. Bibi3, A. Siddiq3, L. Sjstrm2, A. I. F. Blakemore1, P. Froguel1,4, L. M. S. Carlsson2, M. Falchi1; 1 Department of Genomics of Common Disease, School of Public Health, Imperial College London, London, United Kingdom, 2Department of Molecular and Clinical Medicine and Center for Cardiovascular and Metabolic Research, The Sahlgrenska Academy, Gothenburg, Sweden, 3Department of Epidemiology and Biostatistics, School of Public Health, Imperial College London, London, United Kingdom, 4CNRS 8090-Institute of Biology, Pasteur Institute, Lille, France.

Introduction: Obesity is a major contributor to ill health worldwide. Exploration of the genetic contribution to obesity using genome wide approaches has revealed both SNP and CNV associations. Here, we have used a novel approach to analysis of microarray-derived GWAS data to assess the contribution of CNVs to obesity and gene expression. Subjects and Methods: The SOS SibPair cohort consists of 154 nuclear families (732 individuals), each containing a body mass index-discordant sib pair (10

Background: Ethicists suggest that researchers are obliged to report individual genetic research findings to study participants. Opponents contend that promising or reporting provisional knowledge may cause harm and that clinical follow-up regarding these findings may not be available to research participants. Methods: A cross-sectional quasiexperimental survey using vignettes to understand factors that influence researchers judgments about this obligation was administered to cystic fibrosis and autism researchers, internationally. Results: 80% of 342 researchers agreed that individuals in whom a genetic variation is identified should be informed of this finding if judged to be clinically significant. Researchers felt 30-70% less confident that a given finding was clinically significant when it was related to autism research, less scientifically robust, incidental to the index condition, and when barriers to receiving clinical services were perceived (p<.05). Researchers were 30% less likely to report scientifically weaker findings and 40% less likely to report findings when they lacked capacity to provide requisite medical care (p<.05). Compared to molecular and statistical researchers, clinical researchers were 1.8 times more likely to endorse the significance of a given finding and 1.5 times more likely to report it to participants (p<.05). Conclusion: Judgments about reporting genetic research results are influenced not only by scientific parameters, but also by the role of the researcher and his/her capacity to meet par-

Concurrent Sessions
ticipants clinical needs when provisional knowledge is reported. Given the complexity reflected herein, we question the appropriateness of an unequivocal imperative to disclose individual research findings to study participants. c11.3 Direct-to-consumer genetic testing companies: what are their policies regarding testing in minors?
H. C. Howard, P. Borry; KULeuven, Leuven, Belgium.

cancer patients. Objectives: We assessed patients experience and attitudes towards the biobank informed consent process. Methods: A mixed-methods design was used. Nineteen patients (aged 28-82) were in-depth interviewed using grounded theory methodology and 574 patients (aged 20-89, response rate=77.0%) treated for colorectal, breast cancer or a haematological malignancy answered to a self-administered questionnaire. Findings: Two hundred thirteen patients (37.1%) declared they had given consent. Among them less than the half (41.7%) understood that the consent included an authorisation to access to medical files. The interviews pointed out that while contribution to biobanks is considered an act of solidarity and reciprocity, persons were more reluctant to consider financial issues. Most of the patients (82.4%) would accept that a biological sample could be given to another public laboratory. This rate was higher than considering a transfer to a private laboratory (62.9%; p <0.001). Acceptance that tumour samples could be sold were very low for both types of laboratories even more for a public laboratory (10.6% vs 13.2%; p= 0.017). Conclusion: Quality of biobank informed consent should be improved. Specific information should be provided on the potential transfer of samples and associated clinical data for large-scale multicentric research. Education should also stress financial issues in science, for the purposes of transparency and efficacy. c11.6 Biobanking cancer tissue. Patients consider excised (tumour) tissue to be connective tissue.
E. Vermeulen; VUmc, Amsterdam, Netherlands.

Advances in genetic knowledge and technologies have increased the possibilities of testing asymptomatic minors for late-onset diseases, carrier status and susceptibility to common complex disorders. In addition, there has been a recent increase in companies offering genetic tests directly-to-consumers and bypassing the traditional face-to-face encounter with a health care professional from the established healthcare system. With this in mind, we gathered information regarding policies on testing in minors from the websites of 29 companies offering direct-to-consumer (DTC) health-related genetic testing. The results of this study showed that almost half of the companies did not present any information regarding a policy for testing in minors, and that many of the remaining companies had ambiguous statements regarding this issue. Therefore, as a follow up to this content analysis, we sent surveys to 37 companies (including the 29 companies that were part of our initial study) asking them specific questions about their policy on genetic testing in minors. The analysis of survey responses is underway and will provide information not only about companies policies, but also about the actual demand for testing in this population. Given the concerns about the ethical, legal and psychological implications of performing genetic tests in healthy children, it is important to monitor the activities of DTC genetic testing companies in this regard. c11.4* Genetic counsellors Views Regarding their Role in Delivering a Pharmacogenetic service
A. V. E. Callard, W. Newman, K. Payne; University of Manchester, Manchester, United Kingdom.

Currently, there is no clear resolution on which healthcare profession should play a role in delivering a pharmacogenetic service. In addition, there is limited published evidence on healthcare professionals views on pharmacogenetic services. This study aimed to explore the views of genetic counsellors on pharmacogenetic services. Semi-structured face-to-face interviews were carried out with 14 genetic counsellors from two regional genetics clinics in England. The interviews comprised open questions designed to explore their views on potential (dis)advantages of pharmacogenetic testing and the role of counsellors and other professionals in delivering a pharmacogenetic service. In addition, four vignettes describing types of pharmacogenetic tests were used to elicit the counsellors views on specific clinical applications. Data were analysed using the constant comparative method to identify key themes. Opinions on whether genetic counsellors should be involved in a pharmacogenetic service were varied. The genetic counsellors interviewed did see a potential role for their profession but further training may be needed due to their limited knowledge of pharmacogenetics and drug prescription. A regional genetics centre was perceived to be an inappropriate clinical setting for a pharmacogenetic service. However, employing genetic counsellors in a specialist role within a multidisciplinary clinic could be beneficial. Genetic counsellors responses to the vignettes suggested that different pharmacogenetic tests are likely to have different issues, in terms of setting up models of service delivery, and implications for patients. Therefore any clinical service model developed for pharmacogenetic testing will need to be adapted according to the specific issues each test presents. c11.5 informed consent for large-scale biobank research: experience and attitudes of cancer patients

Background Excised tissues are routinely stored in hospitals for future diagnostic use. These tissues are also important for scientific research. In the Netherlands residual tissues are registered in a central pathological registry (PALGA), which makes this collection, consisting of millions of samples, a very large biobank. Aim The presentation elucidates and analyses the underlying attitude of patients towards medical research with excised tissues and focuses on the use of the tissue in genetic medical research. Concepts of ownership in the context of increasing (commercial) value of tissues are discussed. Methods The study is based on mixed methods design combining quantitative data (questionnaires) with qualitative data (transcripts from interviews) and observations during an intervention study at the Netherlands Cancer Institute. In total 260 patients were interviewed, 61% of 426 patients who completed a written questionnaire. Results Many (38%) patients considered themselves to be owner of stored tissue and 43% considered themselves owner of DNA in stored tissue. Most, 98%, endorse medical research with this tissue. For patients the stored tissue is a hypercollective good that should remain in the public sphere in order to facilitate research. Patients consider the tissue to be connective because it connects them to others through research. Patients expect to be reciprocated by the tissue holder and be informed about findings of the research. Conclusion Information about medical research with residual tissue should be improved. A more participatory and reciprocal model of biobanking is required. c12.1 Homozygosity mapping of Primary microcephaly in 112 iranian Families: Novel mutations and Phenotypes
H. Darvish1, S. Esmaeeli Nieh1, G. Bahrami Monajemi1, M. Mohseni1, F. Behjati1, S. Ghasemi Firouzabadi1, I. Bahman1, P. Jamali1, S. Azimi1, F. Mojahedi1, A. Dehghan1, Y. Shafeghati1, A. Jankhah1, M. Falah1, M. Soltani Banavandi1, M. Ghani Kakhki2, M. Garshasbi3, S. Abedini1, s. Banihashemi1, s. arjangi1, F. Rakhshani1, A. Naghavi1, A. Tzschach3, H. Neitzel2, H. Ropers3, A. W. Kuss3, K. Kahrizi1, H. Najmabadi1; 1 GRC, Tehran, Islamic Republic of Iran, 2Institute of Human Genetics, Charit Medical University, Germany, 3Max Planck Institute for Molecular Genetics, Germany.

J. Mancini1,2, C. Chabannon3,4, I. Pellegrini1, F. Viret3,4, N. Vey3,4, C. JulianReynier1; 1 Inserm UMR912 SE4S, Marseille, France, 2Aix-Marseillle Universit, Marseille, France, 3Inserm UMR891 CRCM, Marseille, France, 4Institut Paoli-Calmettes, Marseille, France.

Background: The transfer of biological samples between biobanks is developed to foster the development of large-scale multicentric projects. However, the commercial use of samples and the indispensable concomitant transfer of clinical data associated might be a concern for

Primary microcephaly (MCPH) is a genetically heterogeneous disorder showing an autosomal recessive mode of inheritance. Affected individuals present with head circumferences more than three standard deviations below the age- and sex-matched population mean, accompanied by mental retardation without further associated malformations. Five genes (mcph1, cdk5rap2, aspm, cenpj, STIL) and two genomic

Concurrent Sessions
loci, MCPH2, and 4 have been identified so far. In this study, we investigated all seven loci in patients with primary microcephaly from 112 Iranian families. In addition to a thorough clinical characterization, karyotype analyses were performed for all patients. For linkage analyses, several microsatellite markers were selected for each locus and used for genotyping. Our investigation enabled us to detect linkage to the MCPH5 (ASPM) region in thirteen families. Three families showed linkage to MCPH2, eight to MCPH1 (Microcephalin), five to MCPH6 (CENPJ) and two families were linked to MCPH7 (STIL). The remaining 81 families were not linked to any of the seven known loci. Subsequent sequencing revealed 10 novel mutations and one previously reported mutation in ASPM, 8 novel mutations in MCPH1 as well as a novel mutation in CENPJ. In some families, additional features such as short stature, seizures or congenital hearing loss were observed in the microcephalic patients, which widens the spectrum of clinical manifestations of mutations in known microcephaly genes. Our results show that the molecular basis of microcephaly is more heterogeneous in Iran than elsewhere, thus the Iranian population provides a unique opportunity for finding additional genes underlying this disorder. c12.2* combination of linkage mapping and microarrayexpression analysis identifies NF-B signalling defect as a novel cause for autosomal recessive mental retardation
O. Philippe1, M. Rio1, A. Carioux2, J. Plaza3, P. Guigue1, F. Molinari1, N. Boddaert1, C. Bole-Feysot2, P. Nitschke3, A. Smahi1, A. Munnich1, L. Colleaux1; 1 Hpital Necker-Enfants malades, Paris, France, 2Plateforme de gnomique de la fondation Imagine Paris, Paris, France, 3Plateforme de Bioinformatique de lUniversit Paris Descartes, Paris, France.

novel polymorphisms by genotyping all non-synonymous variants in 300 healthy controls. All but one variant was observed in the controls with allele frequencies of 0.003, 0.005, 0.016. These results indicate that next generation sequencing is a powerful tool to identify mutations associated with rare diseases.
Table 1. Direct identification of a disease-associated mutation by next generation sequencing. carrier Affected Affected carrier mother father child 1 child 2 Number of mapped bases 403,546,905 362,750,092 410,247,834 399,184,755 48.4x 40.5x 47.4x 48.9x mean covarage coverege of targeted regions 99.3% 99.2% 99.3% 99.2% (5) All variants 26,751 27,471 24,115 24,690 10,564 10,694 8,200 8,289 dbsNP129 Novel variants 16,157 16,777 15,915 16,401 High confidence variants Heterozygous sNPs Homozygous sNPs Novel heterozygous variants Novel homozygous variants common variants to all samples G-1 (coding, splice site, RNA) G-2 (5 and 3UtR) G-3 (intronic) G-4 (intergenic) G-1 mendelian compatibility After population screening

4,874 4,647 1,331 1,090

5,076 4,523 1,600 1,602 1555 26 24 929 577 4 1

408 7,095 513 1,789

596 7,563 458 2,029

Autosomal recessive inheritance accounts for nearly 25% of non-syndromic mental retardation (MR) but the extreme heterogeneity of these conditions markedly hampers gene identification. Combining autozygosity mapping and RNA expression profiling in a consanguineous Tunisian family of three MR children with mild microcephaly and white matter abnormalities identified the NIBP/TRAPPC9 gene, which encodes a NF-B inducing kinase (NIK) and IB kinase complex (IKK) binding protein, as a likely candidate. Sequencing analysis revealed a nonsense variant (c.1708C>T, p.R570X) within the exon 9 of this gene responsible for an undetectable level of NIBP protein in patient skin fibroblasts. Moreover, TNF-a stimulation assays showed a defect in IkB degradation, suggesting an impaired NF-B signalling in patient cells. This study provides the first evidence of a NF-B signalling defect in isolated MR. C12.3* Targeted next generation sequencing identifies a mutation associated with cerebellar hypoplasia and mental retardation with quadrupedal locomotion

c12.4* the Arg164X mutation in SAMHD1 leads to a novel variant of Aicardi-Goutires syndrome by modulating cytokine expression

M. du Moulin1, H. Thiele2, K. Barczyk3, C. George1, M. Frosch1, W. Schwindt4, J. Roth3, P. Nuernberg2, F. Rutsch1; 1 University Childrens Hospital, Muenster, Germany, 2Cologne Center for Genomics, University of Cologne, Cologne, Germany, 3Institute for Immunology, Muenster University Hospital, Muenster, Germany, 4Department of Clinical Radiology, Muenster University Hospital, Muenster, Germany.

S. I. Gulsuner1, K. Bilguvar2, M. Tan3, U. Tan4, M. Gunel2, T. Ozcelik1; 1 Department of Molecular Biology and Genetics, Bilkent University, Ankara, Turkey, 2Department of Neurosurgery, Yale University School of Medicine, New Haven, CT, United States, 3Department of Neurology, Baskent University, Ankara, Turkey, 4Department of Physiology, Cukurova University, Adana, Turkey.

We describe the utilization of next generation sequencing in the identification of a mutation associated with a rare and heterogenous Mendelian disorder characterized by cerebellar hypoplasia, mental retardation and quadrupedal locomotion (Uner Tan syndrome). Homozygosity mapping, sequence capture (Nimblegen) of the disease locus on chromosome 17p13 and massively parallel DNA sequencing (454 GS FLX) of two affected and two carrier individuals yielded 362 to 403 million base pairs of sequence information from each sample. Between 96.6%-96.7% of all reads fell within the minimal critical region. Variants common to all samples were selected and binned into four groups (G): variants in coding regions, consensus splices-sites and RNA genes are G-1 (n=26), in 5UTRs & 3UTR s are G-2 (24), in introns are G-3 (929). All remaining variants were classified as G-4 (577) (Table-1). A total of 17 novel exonic variants (four non-synonymous, two synonymous, one 5UTR and ten 3UTR) compatible with the expected autosomal recessive inheritance model of the disease allele was observed (Mendelian compatibility). An exclusion based population screening approach was adopted to distinguish the disease causing variant from

Aicardi-Goutires syndrome (AGS) is a rare inborn multisystemic disease, resembling intrauterine viral infection resulting in psychomotor retardation, spasticity and chilblain-like skin lesions. Diagnostic criteria include intracerebral calcifications and elevated interferon-alpha and pterins in cerebrospinal fluid. Patients present early in infancy and death usually occurs during childhood in a state of decerebration. We report on four adult siblings of Turkish origin with unknown neuro-degenerative disease presenting with stenoses of intracranial vessels, stroke and glaucoma in childhood, two of whom died at the age of 40 and 29 years. Genome wide homozygosity mapping identified 170 candidate genes embedded in a common haplotype of 8Mb on chromosome 20q11-13. Next generation sequencing of the entire region identified the c.490C>T (p.Arg164X) mutation in SAMHD1, a gene most recently described in AGS, on both alleles in all affected siblings. Clinical diagnosis of AGS was then confirmed by demonstrating intracerebral calcifications on cranial computed tomography and elevated pterin levels in cerebrospinal fluid in the two surviving siblings. In patient fibroblasts SAMHD1 protein was undetectable, while basal expression of interleukin-8 was increased and stimulated expression of interferon- was reduced. We conclude that the Arg164X mutation in SAMHD1 by modulating intravascular cytokine expression leads to a novel phenotypic variant of AGS. c12.5 systematic mutation search in families with X-linked mental retardation by next-generation sequencing

V. M. M. Kalscheuer1, W. Chen1,2, S. Haas1, H. Hu1, A. Emde1, C. Menzel1, M. Bienek1, T. Zemojtel1, R. Ullmann1, S. OKeeffe1, M. Vingron1, K. Wrogemann1, A. Tzschach1, A. de Brouwer3, H. van Bokhoven3, N. Lebrun4, M. Raynaud5, H. Van Esch6, H. H. Ropers1;

Concurrent Sessions
Max-Planck-Institute for Molecular Genetics, Berlin, Germany, 2Max-Delbrck Center, Berlin, Germany, 3Department of Human Genetics, Radboud University Medical Centre, Nijmegen, Netherlands, 4INSERM 129-ICGM, Facult de Mdecine Cochin, Paris, France, 5INSERM, U930; Centre Hospitalier Rgional Universitaire de Tours, Service de Genetique, Tours, France, 6Center for Human Genetics, University Hospital Leuven, Leuven, Belgium.
1

c13.1 A recurrent 14q32.2 microdeletion mediated by expanded tGG repeats

X-linked mental retardation (XLMR) affects 1-2/1,000 males and accounts for 10% of all forms of mental retardation. Recent screening of X-linked genes has revealed truncating mutations in 25% of the families studied (Tarpey et al, 2009). Previous studies had indicated that mutations in known XLMR genes account for at least 42% of XLMR families (de Brouwer et al, 2007). To resolve this discrepancy and to shed more light on the molecular causes of XLMR, we have combined genome partitioning techniques and Next Generation Sequencing (NGS) to find the causative gene defect in another 200 families from the European MRX Consortium. Here we report the results on the first 100 families. About 25% of the families carried potentially pathogenic mutations in known XLMR genes. Many other families carried non-synonymous, possibly pathogenic changes in novel genes, but only 3 pathogenic truncating mutations were observed. There are several plausible explanations for this: prior to these investigations, many of these large families had already been screened for mutations in known XLMR genes and positional candidates. Secondly, the EuroMRX cohort is enriched for families with non-syndromic XLMR, and it is conceivable that complete loss of function will often result in severe syndromic forms or even lethality. Thus, the majority of genes whose loss gives rise to non-syndromic XLMR may already be known. Finally, the fundamental defect may not reside in coding regions of the X-chromosome. Sequencing of entire sorted X-chromosomes is being performed to shed more light on these issues. Supported by BMBF and Max-Planck-Society. C12.6 Deep sequencing leads to the identification of 3 independent mutations in the st3GAL3 gene in patients with autosomal recessive intellectual disability from 3 consanguineous iranian families

F. Bna1, S. Gimelli1, E. Migliavacca2, N. Brun-Druc3, K. Buiting4, S. E. Antonarakis1,5, A. Sharp5; 1 Service of Genetic Medicine, University Hospitals of Geneva, Geneva, Switzerland, 2Swiss Institute of Bioinformatics, Department of Genetic Medicine and Development, University of Geneva Medical School, Geneva, Switzerland, 3 Service of General Pediatrics, University Hospitals of Geneva, Geneva, Switzerland, 4Institut fr Humangenetik, Universittsklinikum Essen, Essen, Germany, 5Department of Genetic Medicine and Development, University of Geneva Medical School, Geneva, Switzerland.

Nearly all recurrent microdeletion/duplication syndromes described to date are characterized by the presence of flanking low copy repeats that act as substrates for non-allelic homologous recombination (NAHR) leading to the loss, gain or disruption of dosage sensitive genes. We describe an identical 1.11 Mb heterozygous deletion of 14q32.2 including the DLK1/GTL2 imprinted gene cluster in two unrelated patients. In both patients the deleted chromosome 14 was of paternal origin, and consistent with this both exhibit clinical features compatible with UPD(14)mat. Using a high-resolution oligonucleotide array we mapped the breakpoints of this recurrent deletion to large flanking (TGG)n tandem repeats, each approximately 500bp in size and sharing 88% homology. These expanded (TGG)n motifs share features with known fragile sites and are predicted to form strong guanine quadruplex secondary structures. We suggest that this recurrent deletion is mediated either by NAHR between the TGG repeats, or alternatively results from their inherent instability and/or strong secondary structure. Our results define a recurrent microdeletion of the 14q32.2 imprinted gene cluster mediated by flanking (TGG)n repeats, identifying a novel mechanism of recurrent genomic rearrangement. Our observation that expanded repeats can act as catalysts for genomic rearrangement extends the role of triplet repeats in human disease, raising the possibility that similar repeat structures may act as substrates for pathogenic rearrangements genome-wide. c13.2 Recurrent chromosomal translocations mediated by Genomic interchromosomal NAHR

A. W. Kuss1, H. Hu1, M. Garshasbi1, I. Bahman2, S. Ghadami2, M. M. Motazacker1, L. Abbasi-Moheb1, S. Esmaeeli-Nieh1, L. Puettmann1, M. Mohseni2, M. J. Soltani Banavandi2, H. Darvish2, P. Jamali3, P. Nikoui4, F. Soleimani2, K. Kahrizi2, A. Tzschach1, W. Chen1, H. Ropers1, H. Najmabadi2; 1 Max Planck Institute for Molecular Genetics, Berlin, Germany, 2Genetics Research Center of the University of Social Welfare and Rehabilitation Sciences, Teheran, Islamic Republic of Iran, 3Shahroud Welfare Institution, Sharoud, Islamic Republic of Iran, 4Bandar Abbas Welfare Institution, Bandar Abbas, Islamic Republic of Iran.

S. W. Cheung1, P. Stankiewicz1, A. M. Breman1, J. Wiszniewska1, L. M. Cooper1, S. T. South2, S. Kang1, A. Patel1, J. R. Lupski1, Z. Ou1; 1 Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, United States, 2Department of Pediatrics and Pathology, University of Utah, Salt Lake City, UT, United States.

Most mutations causing intellectual disability (ID) are thought to affect autosomal genes, yet to date only six are known to play a role in the etiology of autosomal recessive ID (ARID). By identifying 12 additional loci in 78 families with non-syndromic ARID (NS-ARID) we have previously shown, that the molecular basis of NS-ARID is extremely heterogeneous (Najmabadi et al., 2007, Hum.Genet. 121(1):43-8). Indeed, only twice have more than one independent NS-ARID causing mutation so far been found to affect the same gene (TUSC3, TRAPPC9). We have now identified three consanguineous families where linkage analysis yielded overlapping homozygous intervals on chromosome 1p34. Using Chromosome sorting to enrich Chr1 from lymphoblastoid cell lines of patients from two of these families followed by deep sequencing, we found two different missense mutations affecting the ST3GAL3 gene. Subsequent screening of ST3GAL3 in the remaining family revealed an additional mutation with a putatively damaging splicing effect. All three changes co-segregate with the disease and were absent in more than 150 controls. This supports the assumption that ST3GAL3 might have a considerably increased mutation frequency in ARID, at least in Iran. The ST3GAL3 gene product is a type II membrane protein, which catalyzes sialic acid transfer from CMP-sialic acid to galactose-containing substrates. Together with colleagues from the Hannover Medical School (M. Muehlenhoff et al.), we are presently investigating the impact of the mutations on ST3GAL3 protein function to understand the pathology of MR in these patients and thus also gain new insights into human brain function.

Sequence analysis of the breakpoints were performed in 4 unrelated families with the same unbalanced translocation der(4)t(4;11)(p16.2;p 15.4) and revealed the presence of large 200-300 kb low copy repeats (LCRs) on 4p16.2 and 11p15.4 of 94% interchromosomal sequence identity. The breakpoints for both the short arms of chromosomes 4 and 11 were mapped within the homologous subunits; consistent with nonallelic homologous recombination (NAHR) as the mechanism for translocation. To investigate the potential involvement of interchromosomal LCRs in recurrent chromosomal translocation formation, we performed computational genome-wide analysis and identified 470 interchromosomal LCRs substrate pairs, greater than 30 kb in size and sharing >94% sequence identity, that may mediate chromosomal translocations. Several predicted recurrent translocations were identified, initially at the G-band level of resolution from clinical cytogenetic databases; in two translocations, t(4;8)(p16.1;p23.1) and t(8;12)(p23.1;p13.31), the breakpoints of seven cases and two cases, respectively, were mapped molecularly to the predicted LCRs. We show that interchromosomal LCRs in 11p15.4 mediate the recurrent constitutional translocation t(4;11)(p16.2;p15.4) via NAHR, provide a computationally determined recurrent translocation map and demonstrate its utility, and further suggest that NAHR may mediate recurrent translocations throughout the human genome. c13.3 the interpretation of copy number gains detected by high resolution array cGH in routine diagnostics; a practical guideline

C. M. A. van Ravenswaaij-Arts, N. M. Hanemaaijer, G. van der Vries, T. Dijkhuizen, R. Hordijk, A. van Essen, B. Leegte, K. Kok, B. Sikkema-Raddatz; Department of Genetics, University Medical Centre Groningen, University of

Concurrent Sessions
Groningen, Groningen, Netherlands.

7
FANCD2 visibly localizes to telomeric foci and PML bodies in ALT, but not in telomerase-positive cells. FANCD2 almost always localizes to telomeric foci containing BLM, and telomeric localization of FANCD2 is BLM-dependent. FANCD2 depletion results in ALT-specific telomere dysfunction characterized by increases in telomeric DNA synthesis, entanglements, recombination events, and association with DNA damage response proteins. Amplified telomeric DNA in FANCD2-depleted cells is primarily extrachromosomal, accumulating both outside of and within PML bodies. We previously reported that BLM overexpression causes a similar phenotype of rapid, large scale ALT-specific amplification of telomeric DNA. Co-depletion of BLM with FANCD2 completely suppresses the telomere phenotypes caused by FANCD2 knockdown, suggesting that the FANCD2-depletion phenotype requires functional BLM. In contrast, co-depletion of RAD51 with FANCD2 does not suppress the FANCD2-depletion phenotype, suggesting that human ALT is RAD51 independent. We propose that FANCD2 restrains BLM-dependent, RAD51-independent recombination and amplification of telomeric DNA in ALT cells by limiting production of ssDNA gapped regions in telomeric DNA, and/or by affecting stability of recombination/replication intermediates. c13.6 Delineating complex genomic architecture involving chromosome segmental duplications
W. A. Khan, J. H. M. Knoll, P. K. Rogan; University of Western Ontario, London, ON, Canada.

The introduction of high resolution array Comparative Genomic Hybridisation (CGH) has led to the detection of large numbers of copy number variations (CNVs) in patients with developmental delay and/or multiple congenital anomalies (DD/MCA) as well as in healthy individuals. The detection of a CNV in DD/MCA does not automatically imply a phenotypic effect. The interpretation of copy number gains is even more complicated than of copy number losses, because of the milder and more variable phenotype associated with microduplications. The aim of the present study was to develop a guideline for the interpretation of gains detected by array-CGH. All copy number gains of at least 4 adjacent oligonucleotides but less than 10 Mb in size, detected in 300 consecutive patients analysed by custom made105K Agilent oligo array were collected and the clinical relevance was evaluated using an interpretation scheme. Of a total of 797 gains, 45% were de novo and 55% were familial. 95% of all de novo and 87% of all familial gains were concluded to be benign CNVs. Fifteen pathogenic gains, sized 288-7,912 kb, were detected. These gains were significantly larger than benign gains and gains of unknown clinical relevance (p<0.001). Surprisingly, they were more often familial (10/14) than de novo (4/14) (not statistically significant). A threshold of 200 kb for the detection of gains in routine diagnostics was concluded to be satisfactory at the moment. A practical guideline to interpret copy number gains was formulated and validated using an independent patient cohort. This guideline will be discussed. c13.4 Paternal origin of de novo constitutional t(11;22)(q23;q11)
T. Ohye1, H. Inagaki1, H. Kogo1, M. Tsutsumi1, T. Kato1, M. Tong1, M. V. E. Macville2, L. Medne3, E. H. Zackai3, B. S. Emanuel3, H. Kurahashi1; 1 Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Japan, 2University of Hospital Maastricht, Maastricht, Netherlands, 3 The Childrens Hospital of Philadelphia, Philadelphia, PA, United States.

The constitutional t(11;22)(q23;q11) is a well-known recurrent nonRobertsonian translocation in humans. While translocations generally occur in a random fashion, the breakpoints of t(11;22)s are concentrated within several hundred base pairs on 11q23 and 22q11. These regions are characterized by palindromic AT-rich repeats (PATRRs), which could cause the genomic instability. De novo t(11;22)s are detected in sperm from healthy males at a frequency of 1/104-105, but never in lymphoblasts, fibroblasts or other human somatic cell lines, suggesting that the generation of a t(11;22) is linked to gametogenesis. However, female germ cells have not been tested since the number of human oocytes that can be examined is limited. To investigate whether the translocation is gametogenesis-specific or male germ cell specific, we attempted to determine parental origin of de novo t(11;22) cases. We studied 8 carriers of de novo t(11;22)s and their parents. The PATRRs and flanking regions on chromosome 11, 22, der(11) and der(22) were amplified by PCR and the nucleotide sequences were determined. The highly polymorphic nature of the PATRRs allowed us to determine the parental origin of the de novo t(11;22)s. All of the eight translocations were found to be of paternal origin. This result implicates a possible novel mechanism of sperm-specific generation of t(11;22)s. It is proposed that replication errors during the numerous cell divisions in pre-meiotic spermatogenic cells or conformational changes of the DNA during chromatin remodeling in the post-meiotic stages of spermatogenesis might contribute to male-specific formation of de novo t(11;22)s. c13.5 ALt-immortalized human cells are critically dependent on the Fanconi anemia protein FANcD2 to limit BLm-dependent recombination and amplification of telomeric repeat DNA
H. Root, A. Larsen, D. Bazett-Jones, S. Meyn; Hospital for Sick Children, Toronto, ON, Canada.

Complex genomic architectures predispose to chromosomal rearrangements. The rearrangements can cause a diverse range of phenotypes due to haploinsufficiency or positional effects from genes in cis with families of segmental duplications (SDs). Genomic regions flanked by SDs in Angelman and Prader-Willi syndromes (AS/PWS) show a great proclivity for such chromosomal structural changes. The most common rearrangements in AS/PWS involve large (Class I) and small (Class II) deletions sharing a common telomeric region of breakage. FISH can detect juxtaposed DNA sequences in normal and abnormal chromosomes which can delineate the locations of these sequences within individual SD elements based on differences in their respective genomic contexts. In this study, we computationally designed and developed 47 single copy (sc) and low copy (lc) sequence-defined FISH probes to delineate proximal and distal ends of the chromosome 15q11-q13 deletion in Class I and Class II deletions. Departure from the expected fluorescent signals using a two-step hybridization algorithm directed which sc and lc FISH probe combinations were used in subsequent hybridizations. In situ analysis demonstrated variations in deletion extent at both the proximal and distal ends within each patient group. Delineated genomic intervals suggest that SDs of both high and low sequence homologies as well as regions adjacent to SD blocks are giving genesis to the dynamic breakage activity in this highly unstable region. These strategies should be useful for delineation of rearrangement boundaries for other disorders associated with SDs and can provide unifying mechanisms by which complex genomic architectures cause instability in human chromosomes. c14.1 Dysostin, a new gene involved in cDG and affecting pH homeostasis
F. Foulquier1, J. Jaeken2, M. Amyere3, R. Zeevaert2, L. Keldermans4, M. Vikkula3, E. Van Schaftingen3, G. Matthijs4; 1 Universit des Sciences et Technologies de Lille, Lille, France, 2Center for Metabolic Disease, University of Leuven, Leuven, Belgium, 3de Duve Institute, Universit Catholique de Louvain, Brussels, Belgium, 4Center for Human Genetics, University of Leuven, Leuven, Belgium.

Fanconi anemia (FA) is an inherited disorder characterized by bone marrow failure, cancer predisposition and congenital malformations. FA proteins are implicated in homologous recombination (HR), a process involved in Alternative Lengthening of Telomeres (ALT) pathways. The FA protein FANCD2 interacts with other HR proteins, however the precise role of FANCD2 in HR is unclear. We now find FANCD2s ability to limit BLM-dependent telomeric recombination and amplificiation events is critical for human ALT cell telomere maintenance and survival.

The correct structure of the oligosaccharides on glycoproteins depends on the enzymatic activities of numerous glycosyltransferases and remodeling glycosidases, but also on a precise Golgi localization of these enzymes and an adequate intracellular environment. Genetic defects affecting this glycosylation pathway cause a range of diseases known as Congenital Disorders of Glycosylation (CDG). Through a combination of homozygosity mapping and expression analysis in fibroblasts, we have been able to identify a novel gene involved in CDG in a single, nuclear family, and pinpoint the defect to a deep intronic mutation. We subsequently identified missense mutations in 2 other, unrelated cases. Because of the patients phenotype, we named it dysostin. In the patients fibroblasts, a dilated Golgi morphology associated with

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a fragmentation of the trans Golgi network was observed. A slight delay in the retrograde translocation of Golgi membranes to the endoplasmic reticulum was detected after treatment with brefeldin A, suggesting that intracellular trafficking is compromised. Co-localization of the fluorescently-tagged protein with relevant markers revealed a late endosomal/lysosomal localization, which is peculiar in view of the glycosylation defect. pH measurements in late endocytic structures using the Lysosensor dye revealed a higher staining - and hence a more pronounced acidity - in those compartments in the patients. This result was reproducible in Hela cells, using a siRNA strategy to knockdown dysostin. In summary, the powerful combination of genetic and expression studies allowed us to discover a novel CDG gene; the cellular assays revealed completely novel connections between lysosomal pH, glycosylation and intracellular trafficking. c14.2 A new inborn error of glycosylation due to DPm2 deficiency

the ten annotated genes in the candidate region revealed homozygosity for a single-nucleotide deletion at position c.-95 in the proteasome maturation protein (POMP) gene, in all patients. The deletion was associated with a shift in expression of POMP transcript variants with a marked increase of transcripts with elongated 5 untranslated regions (UTRs) in keratinocytes. POMP functions as a chaperone for proteasome maturation and immunohistochemical analysis of skin biopsies from KLICK patients revealed an altered distribution of POMP, the proteasome subunit proteins 7 and 5 and the ER stress marker CHOP in the most differentiated skin layers. Furthermore, the KLICK patients showed a deviant expression of the skin differentiation marker filaggrin. Our results suggest that KLICK syndrome is caused by a single-nucleotide deletion in the 5 UTR of POMP, resulting in altered distribution of POMP and proteasomes in epidermis and a perturbed formation of the outermost layers of the skin. These findings imply that the proteasome has a prominent role in the terminal differentiation of human epidermis and that elevated ER stress is a disease mechanism in genodermatoses. c14.4* mutation in SHOC2 promotes aberrant protein Nmyristoylation and underlies Noonan-like syndrome with loose anagen hair

V. Race1, R. Bammens1, W. Vleugels1,2, R. Barone3,4, A. Fiumara4, F. Foulquier2, G. Matthijs1; 1 Center for Human Genetics, University of Leuven, Leuven, Belgium, 2Unit de Glycobiologie Structurale et Fonctionnelle UMR/CNRS 8576, IFR147, Lille, France, 3Department of Neuroscience, University of Catania, Catania, Italy, 4 Department of Pediatrics, University of Catania, Catania, Italy.

Congenital Disorders of glycosylation (CDG) are a group of complex metabolic diseases. About 40 types have been identified. Most CDG with a type I pattern on transferrin isolectric focusing (CDG-I) result from defects in the glycosyltransferases and sugar transporters, needed for glycan assembly in the endoplasmic reticulum. We report on a CDG-I patient with severe developmental delay, epilepsy and dysmorphic features,. Analysis of the lipid-linked oligosaccharides (LLO) showed an accumulation of dol-PP-GlcNAc2-Man5. This pattern is compatible with a defect in DPM1, ALG3 or MPDU1, in which mutations have been described; however, no mutations were detected. We decided to further analyze the DPM2 and SAC1 genes. The patient was found to be compound heterozygote for 2 mutations in DPM2: a splice mutation (c.4-1G>C) and a missense mutation (c.68A>G, p.Y23C). Hence, this patient presents with a novel type of CDG.The human dolichol-phosphate-mannose (DPM) synthase is a heterotrimeric complex composed of DPM1, DPM2 and DPM3. Until now, only mutations in DPM1 (the catalytic subunit) and more recently in DPM3 have been described. DPM2 is a hydrophobic protein of 84 amino acids, whose function is still not clear. It is reported in the literature that DPM2 could be involved in the regulation of the DPM synthase complex but also in the regulation of the glycosylphosphatidylinositols-N-acetylglucosaminyltransferase, and a defect in DPM2 may also affect GPI-anchored proteins - a feature which has not been studied so far in CDG patients. Further investigation of the DPM2 mutations by complementation in Lec15 hamster DPM2-deficient cells is ongoing. c14.3* A single-nucleotide deletion in the POMP 5 UtR causes a transcriptional switch and an altered epidermal proteasome distribution in KLicK genodermatosis

V. Cordeddu1, E. Di Schiavi2, A. Maayan3, A. Sarkozy4,5, V. Fodale1, S. Cecchetti1, G. Zampino6, L. Mazzanti7, M. C. Digilio8, S. Martinelli1, E. Flex1, F. Lepri4, D. Bartholdi9, K. Kutsche10, G. B. Ferrero11, A. Selicorni12, C. Anichini13, C. Rossi14, R. Tenconi15, M. Zenker16, B. Dallapiccola4, R. Iyengar3, P. Bazzicalupo2, B. D. Gelb3, M. Tartaglia1; 1 Istituto Superiore di Sanit, Rome, Italy, 2Istituto di Genetica e Biofisica A. Buzzati Traverso, Naples, Italy, 3Mount Sinai School of Medicine, New York, NY, United States, 4IRCCS-CSS Istituto Mendel, Rome, Italy, 5Institute of Human Genetics, Newcastle University, Newcastle upon Tyne, United Kingdom, 6 Universit Cattolica del Sacro Cuore, Rome, Italy, 7Universit di Bologna, Bologna, Italy, 8Ospedale Bambino Ges, Rome, Italy, 9Institute of Medical Genetics, University of Zurich, Zurich, Switzerland, 10Universittsklinikum Hamburg-Eppendorf, Hamburg, Germany, 11Universit di Torino, Turin, Italy, 12 IRCCS Fondazione Policlinico Milano, Milan, Italy, 13Universit di Siena, Siena, Italy, 14Policlinico S. Orsola-Malpighi, Bologna, Italy, 15Universit di Padova, Padua, Italy, 16University of Erlangen-Nuremberg, Erlangen, Germany.

J. Dahlqvist1, J. Klar1, N. Tiwari2, J. Schuster1, H. Trm3, J. Badhai1, R. Pujol4, M. van Steensel5, T. Brinkhuijzen5, L. Gijezen5, A. Chaves6, G. Tadini7, A. Vahlquist3, N. Dahl1; 1 Department of Genetics and Pathology, Uppsala University, Uppsala, Sweden, 2Department of Biomedicine, Institute of Biochemistry and Genetics, Universtiy of Basel, Basel, Switzerland, 3Department of Medical Sciences, Uppsala University and University Hospital, Uppsala, Sweden, 4Department of Dermatology, Hospital del Mar IMAS, Barcelona, Spain, 5Department of Dermatology, University Hospital Maastricht, and the GROW school for oncology and developmental biology, University of Maastricht, Maastricht, Netherlands, 6Department of Dermatology, University Hospital Infanta Cristina, Badajoz, Spain, 7Institute of Dermatological Sciences, Fondazione Ospedale Maggiore Policlinico, Milan, Italy.

N-myristoylation is a common form of co-translational protein fatty acylation resulting from the attachment of myristate to a required Nterminal glycine residue. We show that aberrantly acquired N-myristoylation of SHOC2, a leucine-rich repeat-containing protein that positively modulates RAS-MAPK signal flow, underlies a clinically distinctive condition of the neuro-cardio-facial-cutaneous disorders family. Twenty-five subjects with a relatively consistent phenotype previously termed Noonan-like syndrome with loose anagen hair [OMIM 607721] shared the 4A>G missense change (Ser2Gly) in SHOC2 that introduces an N-myristoylation site, resulting in aberrant targeting of SHOC2 to the plasma membrane and impaired translocation to the nucleus upon growth factor stimulation. Expression of SHOC2S2G in vitro enhanced MAPK activation in a cell type-specific fashion. Induction of SHOC2S2G in Caenorhabditis elegans engendered protruding vulva, a neomorphic phenotype previously associated with aberrant signaling. These results document the first example of an acquired N-terminal lipid modification of a protein causing human disease. c14.5* Another gene for autosomal recessive ALX-related frontonasal dysplasias: Disruption in ALX1 (CART1) causes anophthalmia and severe facial clefting

KLICK syndrome is a rare autosomal recessive skin disorder characterized by palmoplantar keratoderma, linear hyperkeratotic papules over joints, sclerotic constrictions around fingers and ichthyosiform scaling. We identified twelve cases, from five European countries, that share the specific KLICK manifestations. Using genome-wide SNP-based homozygosity mapping we identified a 1.5-Mb homozygous candidate region on chromosome 13q. Sequence analysis of

E. Uz1, Y. Alanay2, D. Aktas1,2, I. Vargel3,4, S. Gucer5, G. Tuncbilek4, F. von Eggeling6, E. Yilmaz7, O. Deren8, N. Posorski6, H. Ozdag9, T. Liehr6, S. Balci2, M. Alikasifoglu1,2, B. Wollnik10,11, N. A. Akarsu1; 1 Department of Medical Genetics, Gene Mapping Laboratory, Hacettepe University, Ankara, Turkey, 2Pediatric Genetics Unit, Department of Pediatrics, Hacettepe University, Ankara, Turkey, 3Department of Plastic and Reconstructive Surgery, Kirikkale University, Kirikkale, Turkey, 4Department of Plastic and Reconstructive Surgery, Hacettepe University, Ankara, Turkey, 5Pathology Unit, Department of Pediatrics, Hacettepe University, Ankara, Turkey, 6Jena University Hospital, Institute of Human Genetics and Antropologie, Jena, Germany, 7Department of Medical Biology, Hacettepe University, Ankara, Turkey, 8Department of Obstetrics and Gynocology, Hacettepe University, Ankara, Turkey, 9Biotechnology Institute of Ankara University, Ankara, Turkey, 10Center For Molecular Medicine, University of

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Cologne, Cologne, Germany, 11Institute of Human Genetics, University of Cologne, Cologne, Germany.

c15.1* Genome-wide association study of regional brain volume suggests involvement of known psychiatry candidate genes, identifies new candidates for psychiatric disorders and points to potential modes of their action
A. Arias-Vsquez1, B. Franke1, J. Veltman2, H. Brunner2, P. Hagoort3, G. Fernandez3; 1 Radboud University Nijmegen Medical Center, Departments of Psychiatry & Human Genetics, Nijmegen, Netherlands, 2Radboud University Nijmegen Medical Center, Department of Human Genetics, Nijmegen, Netherlands, 3 Donders Centre for Cognitive Neuroimaging, Donders Institute for Brain, Cognition and Behavior, Radboud University Nijmegen (Medical Centre), Nijmegen, Netherlands.

Molecular genetics of recessively inherited frontonasal dysplasias (FNDs) are largely unknown. In 2009, two studies have drawn attention to the critical role of aristales-related homeobox transcription factors, ALX3 and ALX4, in the molecular pathogenesis of autosomal recessive-FND in humans (Am. J. Hum. Genet. 2009; 84: 698 and Hum Mol Genet. 2009; 18: 4357, respectively). We present a new autosomal recessive frontonasal dysplasia in two families characterized by bilateral anophthalmia/microphthalmia, bilateral oblique facial cleft, complete cleft palate, hypertelorism, wide nasal bridge with hypoplasia of ala nasi, and low set and posteriorly rotated ears. Using Affymetrix 250K SNP Array genotyping and homozygosity mapping, we mapped this clinical entity to chromosome 12q21. In one of the families with three affected sibs, CNV analysis of the critical region detected a homozygous 3.7 Mb deletion containing the ALX1 (CART1) gene. In the second family, we identified a homozygous donor splice site mutation (c.537+1 G>A), which is predicted to disrupt the functionally essential homeodomain structure of the ALX1 protein. These results provide evidence that loss of ALX1 function causes severe impairment of early craniofacial development. Unlike its murine ortholog, complete loss of human ALX1 does not result in neural tube defects, however, severely affects the orchestrated fusions between frontonasal, nasomedial, nasolateral, and maxillary processes in early embryogenesis. This study further expands the spectrum of the recently recognized autosomal recessive ALX-related FND phenotypes in humans. The study was done within the CRANIRARE consortium supported by the European Research Area Network, E-RARE through Turkish Scientific Council. c14.6* High-throughput mutation screening in combination with cellular complementation of rare variants aid gene identification in mitochondrial disorders

K. Danhauser1, F. Madignier1, M. Herzer1, T. Haack1, B. Haberberger1, P. Freisinger2, B. Rolinski3, R. Horvath2, H. Mayr4, W. Sperl4, V. Tiranti5, M. Tesarova6, B. Plecko7, S. Biskup8, D. Boehm8, A. Giovanetti5, B. Garavaglia5, M. Zeviani5, T. Meitinger1, H. Prokisch1; 1 Institute of Human Genetics, Technical University of Munich and Helmholtz Zentrum, Mnchen, Germany, 2Technical University of Munich, Stoffwechselzentrum Kinderklinik, Mnchen, Germany, 3Klinikum Mnchen GmbH Medizet Stoffwechselzentrum, Mnchen, Germany, 4Universitt Salzburg, Kinderklinik, Salzburg, Austria, 5Division of Molecular Neurogenetics, National Neurological Institute, C. Besta, Milano, Italy, 6University of Prague, Department of Paediatrics, Prag, Czech Republic, 7Department of Pediatrics, Medical University Graz, Graz, Austria, 8CeGaT GmbH, Tbingen, Germany.

Psychiatric diseases are highly heritable disorders with complex etiology partially explained by combinations of genes with small effect size interacting with each other and with environmental factors. The small effect sizes of genetic risk factors have hampered the identification of genes predisposing to these diseases. A possible way to aid identification of risk genes is the use of intermediate phenotypes. These can be defined e.g. at the level of brain function and of regional brain structure. Both are highly heritable, and regional brain structure is linked to brain function. Within the Brain Imaging Genetics (BIG) study at the Radboud University Nijmegen (Medical-Centre) we performed a genome-wide association study (GWAS) in 600 of the currently 1400 healthy study participants. For all BIG participants, structural MRI brain images were available. Gray and white matter volumes were determined by brain segmentation using SPM software. FSL-FIRST was used to assess volumes of specific brain structures. Genotyping was performed on Affymetrix 6.0 arrays. Known candidates from studies on psychiatric genetics and mental disorders are implicated in the regulation of regional brain structure. E.g., CDH13, associated with ADHD, schizophrenia and substance abuse, was associated with amygdala, hippocampus and white matter volumes (P<10E-05); CANA1C, associated with bipolar disorder, was found associated with brainstem volume (p<10E-05) and three SNPs on the PLXNA2 gene showed p-values <10E-05 with caudate nucleus volume. Our data suggests that genes involved in psychiatric and mental disorders are also associated with the variance of intermediate phenotypes based on brain volumes from healthy individuals. c15.2 NXF/ARNt2/sim2, RET Expression regulation and specific HscR Associated DNA Variants
R. M. W. Hofstra1, Y. Sribudiani1, M. Metzger2, J. Osinga1, A. Rey1; 1 UMCG, RUG, Groningen, Netherlands, 2University Leipzig, Leipzig, Germany.

Mitochondria are key players in the maintenance of cellular energy metabolism through the process of oxidative phosphorylation. This task requires five multimeric respiratory chain complexes, with complex I with 45 subunits being the largest one. Complex I deficiency is a major cause of mitochondrial diseases, presenting with an array of phenotypic features and the elucidation of the molecular correlate remains a challenge. We performed a multi-centre large-scale mutation screen of 70 candidate genes in 150 patients with isolated complex I deficiency by high resolution melting curve analysis and Sanger sequencing. For the first time we identified pathogenic mutations in the mitochondrial respiratory chain complex I subunit NDUFB9. Additionally, we identified causative mutations in another 16 genes, which have previously been associated with complex I deficiency: 5 mtDNA encoded subunits, 3 tRNA genes, 6 nuclear encoded subunits and 3 assembly factors. The causality of newly identified missense mutations was established by complementation of complex I activity in patient-derived fibroblast cell lines by lentiviral expression of wildtype cDNA. We have initiated an exome sequencing approach starting with familial cases to identify mutations in the 70% of patients with no mutations found yet. Taking advantage of the complementation approach, we will be able to discriminate pathogenic from non-pathogenic variants.

Two non-coding RET variations, the T allele of SNP rs2435357 (Enh1T) and the A allele of SNP rs2506004 (Enh2-A) proved strongly associated with Hirschsprung (HSCR) susceptibility. Furthermore, in two SNPs are in strong linkage disequilibrium and both located in an enhancer element in intron 1 of the RET gene. For Enh1 it has been shown that the disease associated T allele results in reduced expression in Luciferase experiments, via reduced SOX10 binding, when compared to non-disease associated C allele. The goals of this study were to determine whether Enh2-A also is a functional variant, i.e. affect RET expression. We generated reporter constructs containing both alleles of the two SNPs, separately or in combinations, coupled to the Luciferase gene. Luciferase assays showed that not only the Enh1-T allele but also the Enh2-A allele decreased reporter gene, showing that both SNPs do contribute independently. MatsInspector software identified the sequences of Enh2-C (non-disease associated variant) and its surroundings sequences (-ACGTG-) as a potential binding site for the (heterodimer) transcripton activator NXF/ARNT2, and the (heterodimer) transcription repressors SIM2/ARNT2. Binding affinity of NXF/ARNT2 to Enh2-C was confirmed by Electrophoresis Mobility Shift Assays and Supershift. Transfections of NXF/ARNT2 or SIM2/ARNT2 into Neuroblastoma cell lines increase and decrease RET expression respectively as expected. Interestingly SIM2 is located on chromosome 21 and trisomy 21 strongly increases the risk on HSCR. Most importantly, our data shows that more than one SNP on an associated haplotype might influence disease development making polygenic diseases even more complex than originally thought

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c15.3 Genomewide association of single nucleotide polymorphisms with subcutaneous adipose tissue gene expression.

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c15.5 Functional analysis of fatty acid desaturase (FADs) gene cluster polymorphisms

Exploration of the genetic control of gene expression in adipose tissue could provide valuable insights into the genetic architecture of common human obesity. Using whole genome microarrays, a genome wide association analysis was carried out using gene expression levels measured in subcutaneous adipose tissue samples from obesitydiscordant sibpairs as quantitative traits. The SOS SibPair cohort was utilised, which consists of 154 nuclear families (n=732), each containing an obesity-discordant sib pair (Body Mass Index difference 10 kg/m2). RNA was extracted from subcutaneous adipose tissue and DNA was extracted from blood. Gene expression and genotyping data was generated from all 349 siblings using the Affymetrix Human U133 Plus 2.0 and Illumina Human 610K arrays, respectively. Analysis of the genomewide gene expression and genotype data was carried out using Merlin. Approximately 10% of all transcripts (~5500) were associated with SNP markers in the genome, either in cis or in trans, using the very stringent threshold of a Bonferroni correction of p=3.6 x 10-12. For a False Discovery Rate of 10%, 4400 transcripts differentially expressed between lean and obese siblings were detected. Using a Bonferroni correction of p=1x10-7, approximately 350 of the differentially expressed transcripts were also cis-eQTLs and approximately 150 were also trans-eQTLs. In the next stage, differential expression analysis and incorporation of the obesity status in the association framework will allow the identification of SNP markers and pathways involved in obesity susceptibility. c15.4 mutations of VANGL1 in patients with neural tube defects

A. J. Walley1, M. F. Falchi1, J. Andersson1, P. Jacobson2, A. Bibi1, M. Jarnas2, A. Siddiq1, L. Sjostrom2, L. M. S. Carlsson2, P. Froguel1,3; 1 Department of Genomics of Common Disease, School of Public Health, Imperial College London, London, United Kingdom, 2Department of Molecular and Clinical Medicine and Center for Cardiovascular and Metabolic Research, The Sahlgrenska Academy, Gothenburg, Sweden, 3CNRS 8090-Institute of Biology, Pasteur Institute, Lille, France.

E. Lattka1, S. Eggers1, G. Moeller2, K. Heim3, M. Weber3, D. Mehta3, H. Prokisch3,4, J. Adamski2,5, T. Illig1; 1 Helmholtz Zentrum Mnchen, Institute of Epidemiology, Neuherberg, Germany, 2 Helmholtz Zentrum Mnchen, Institute of Experimental Genetics, Neuherberg, Germany, 3Helmholtz Zentrum Mnchen, Institute of Human Genetics, Neuherberg, Germany, 4Technische Universitt Mnchen, Institute of Human Genetics, Munich, Germany, 5Technische Universitt Mnchen, Lehrstuhl fr Experimentelle Genetik, Freising-Weihenstephan, Germany.

Fatty acid desaturases play a pivotal role in the endogenous formation of n-6 and n-3 long-chain polyunsaturated fatty acids (LC-PUFAs). Several studies reported associations of single nucleotide polymorphisms (SNPs) in the human desaturase encoding genes (FADS1, FADS2) with LC-PUFA levels, with an amazingly high genetically explained variance (28.5%) for arachidonic acid. Moreover, SNPs in this cluster were associated with more complex phenotypes such as cholesterol and triglyceride levels as well as glucose levels. The functional relevant SNP(s) could not be identified by these association studies, because all associated SNPs are in high linkage disequilibrium. The aim of our study is therefore to identify the causative variant(s) to learn more about the regulatory pathways involved in LC-PUFA homeostasis. We identified two interesting polymorphisms (rs3834458 and rs968567) in the FADS2 gene promoter by bioinformatics analysis. Luciferase reporter gene assays revealed that the minor T allele of SNP rs968567 leads to a significant increase in promoter activity in cell lines, whereas rs3834458 showed no significant effect. Competitive electrophoretic mobility shift assays showed allele-specific binding of nuclear proteins for the rs968567 surrounding region, but no differential results were obtained for rs3834458. One of the proteins binding to the rs968567 surrounding region in an allele-specific manner was identified as transcription factor ELK1. Altogether, our results indicate that rs968567 influences FADS2 transcription, probably by genotype dependent ELK1 binding, and offer first insights into modulation of FADS2 gene transcription by SNPs. c15.6 mutation and functional analysis of the iRAK-m gene in asthmatic patients.

P. De Marco1, C. M. Bosoi2, E. Merello1, A. Reynolds2, S. Lachance2, J. R. McDearmid3, P. Gros4, A. G. Bassuk5, A. Cama1, K. Zoha2, V. Capra1; 1 Neurosurgery Department, G. Gaslini Institute, Genoa, Italy, 2CHU Sainte Justine Research Center, Department of Obstetrics and Gynecology, University of Montreal, Montreal, QC, Canada, 3The Center for Research in Neuroscience, Research Institute of the McGill University Health Center, and Department of Pathology and Cell Biology, University of Montreal, Montreal, QC, Canada, 4 Department of Biochemistry, McGill University, Montreal, QC, Canada, 5 Department of Pediatrics, University of Iowa, Iowa, IA, United States.

S. Sanna1,2, C. A. Caria1, F. Anedda1, A. Loi1, F. Virdis1, N. Olla1, L. Balaci1, G. Sole1, M. Uda1, S. Naitza1; 1 Istituto di Neurogenetica e Neurofarmacologia, Consiglio Nazionale delle Ricerche, Monserrato, Monserrato, Italy, 2Regione Autonoma della Sardegna, Italy.

Neural tube defects (NTD) are congenital malformations resulting from failure of neurulation. Recent years witnessed a breakthrough in elucidating the role of planar cell polarity (PCP) pathway in neurulation and how molecular lesions in this pathway lead to NTD in animal models and humans. PCP is controlled by the non canonical Fz/Dvl signalling pathway that involves a number of additional core gene, including Stbm/Vang, Fmi, Pk, and Dgo. The Loop-tail (Lp) mouse that develops craniorachischisis carry missense mutations in the Vangl2 gene, that is the mammalian homolog of the Drosophila Stbm/Vang gene required for establishing planar cell polarity in many tissues. This mouse provided the first line of evidence for involvement of PCP pathway in NTD in mammals. Vangl1, a vertebrate homolog of Vangl2, encodes for a transmembrane protein containing a PDZ-domain binding motif involved in protein-protein interactions. We sequenced human VANGL1 gene in a cohort of 810 NTD patients and we identified 8 missense mutations both in familial (V239I, R274Q, S83L, and R181E) and sporadic (M238T, F153S, L202F, and A404S) cases. These mutations affect highly conserved residues and were not found in 1200 controls. We demonstrated that V239I mutation abrogates in vitro the interaction between VANGL protein and DVL, strongly suggesting a pathogenic effect on the protein function. Finally, we demonstrated that two human mutations, V239I and M238T, affect convergent extension in zebrafish and we hypothesize that they most likely affect a similar process in humans. These results support a role for VANGL1 as genetic risk factor for NTD.

Asthma is a complex disease caused by the interaction between genes and environment. Since its prevalence and associated mortality is largely increasing in the latest years, there is a great interest in understanding the molecular basis of this pathology. In 2007 we identified IRAK-M as an asthma susceptibility gene in the Sardinian population and replicated the study in two genetically distant populations. To better understand the role of IRAK-M in the pathogenesis of asthma, we first conducted a mutation screen in affected individuals and in controls to identify the functional variations responsible of the association. Our analysis revealed a clustering of rare coding mutations in patients and functional studies in different cell lines showed that some affect downstream activation of NF-B, a transcription factors involved in innate immunity and inflammation negatively regulated by IRAK-M. Combining bioinformatics analysis and deletion studies in cell lines, we identified a region upstream of IRAK-M with promoter activity, containing several binding sites for transcription factors known to modulate inflammatory responses (c-REL, AP1, NFAT). EMSA experiments for the predicted binding sites demonstrated specific binding in monocyte cell lines stimulated with LPS, whereas site directed mutagenesis showed that destruction of the binding sites greatly reduced activity of the IRAK-M promoter region. These data further support a potential role of IRAK-M in the pathogenesis of chronic inflammation and asthma.

Concurrent Sessions
c16.1 the revised Ghent nosology for the marfan syndrome (mFs)
B. L. Loeys1, J. De Backer1, B. Callewaert1, H. Dietz2, A. De Paepe1; 1 Center for Medical Genetics, Gent, Belgium, 2Johns Hopkins University, Baltimore, MD, United States.

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GVMs, somatic UPID may have wide implications. c16.3 Lethal Vascular syndrome from south india due to a Novel mutation in Fibulin 4

The diagnosis of MFS relies on a set of international criteria, outlined by expert opinion. In 1996, the initial Berlin nosology was redefined because of the risk of overdiagnosis into the Ghent nosology, a more stringent set of major and minor criteria. These Ghent criteria have proven to work well since with improving molecular techniques, confirmation of the diagnosis is possible in over 95% of patients. However, concerns with the Ghent criteria are that some of the diagnostic manifestations have not been validated as thresholds and others necessitate cumbersome imaging studies. Moreover, in the absence of aortic dilation, the diagnosis can be stigmatizing, hamper career aspirations and restrict life-insurances opportunities. The label MFS may cause psychosocial burden by restricted exercise permission. Following an international expert meeting, we propose a revised Ghent nosology in which aortic root aneurym and ectopia lentis are cardinal features. In absence of family history, the presence of these two manifestations is sufficient for the unequivocal diagnosis of MFS. In absence of any of these two, the presence of bonafide FBN1 mutation or a combination of systemic features is required. For the latter a new scoring system has been designed and validated. In this way FBN1 testing is not mandatory but useful when available. The proposed new nosology puts more weight on the cardiovascular manifestations of the disease. We anticipate that the new nosology can delay a definitive diagnosis of MFS but decreases the risk of premature or mis-diagnosis and facilitates discussion of risk and management guidelines. c16.2 Acquired uniparental isodisomy as a common somatic 2nd-hit explains multifocality of glomuvenous malformations

S. Nampoothiri1, M. Kappanayil1, A. De Paepe2, B. Loeys2, L. Van Laer2, R. Kannan1, M. Faiyaz-Ul-Haque3, R. Krishna Kumar1; 1 Amrita Institute of Medical Sciences and Research Center, Cochin, Kerala, India, 2Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium, 3 King Faizal Hospital, Saudi Arabia.

M. Vikkula1, V. Aerts1, F. Duhoux2, P. Brouillard1, M. Wassef3, O. Enjolras4, J. Mulliken5, H. Antoine-Poirel2, O. Devuyst6, L. Boon7, M. Amyere1; 1 Laboratory of Human Molecular Genetics, de Duve Institute, Universit catholique de Louvain, Brussels, Belgium, 2Center for Human Genetics, Cliniques universitaires Saint-Luc, Universit catholique de Louvain, Brussels, Belgium, 3Service danatomie et de cytologie pathologiques, Hpital Lariboisire, Paris, France, 4Pediatric Vascular Clinic, Departement of Maxillofacial and Plastic Surgery, Hpital denfants Armand Trousseau, Paris, France, 5Vascular Anomalies Center, Division of Plastic Surgery, Childrens Hospital and Harvard Medical School, Boston, MA, United States, 6Unit of nephrology, Universit catholique de Louvain, Brussels, Belgium, 7Centre for Vascular Anomalies, Division of Plastic Surgery, Cliniques Universitaires SaintLuc, Universit catholique de Louvain, Brussels, Belgium.

We report a lethal disorder characterized by a distinct phenotype and arterial tortuosity in a unique cohort of sixteen infants, caused by a novel mutation in the Fibulin 4 gene. Methods: Prospective, hospital-based study (Jan 2006 - Dec 2009)Results: Sixteen children (9M / 7F) were identified with arterial tortuosity and a distinct phenotype, characterized by long philtrum and thin upper lip (87%), cutis laxa (53%), sagging cheeks (47%), hypertelorism (53%),and long fingers(40%). All children presented with early onset respiratory symptoms (median age 1.5 months) and belonged to unrelated families from the same geographical (South India) background with a history of third degree consanguinity in seven families. Cardiovascular features were documented by echocardiography,cardiac CT and MRI which revealed marked dilatation and tortuosity of aorta and its branches. Thirteen patients died between 36 hours -17 months of age (median 2.5 months) due to respiratory failure. Genetic studies led to identification of a novel homozygous c.608A>C (p.Asp203Ala) mutation in exon 7 of the Fibulin 4 gene in 15/16 patients. In the only patient surviving to the age of 6 years, compound heterozygosity was found for this mutation with a c.679C>T (p.Arg227Cys) mutation. Prenatal study in one couple identified an affected foetus. Haplotype analysis revealed a shared haplotype with Fibulin 4 gene, which strongly suggested common ancestry for all probands due a relatively old founder mutation. Conclusions: This is the first complete description of this lethal genetic disorder and illustrates that Fibulin 4 is critical to human elastogenesis and vascular integrity. c16.4 A new locus for a syndromic form of thoracic Aortic Aneurysms maps to chromosome 15q

I. M. B. H. van de Laar1, R. A. Oldenburg1, B. de Graaf1, J. A. M. Verhagen1, Y. M. Hoedemaekers1, I. Frohn-Mulder2, J. Roos-Hesselink3, J. M. Kros4, B. A. Oostra1, M. W. Wessels1, A. M. Bertoli-Avella1; 1 Department of Clinical Genetics, Erasmus Medical Center, Rotterdam, Netherlands, 2Department of Pediatric Cardiology, Erasmus Medical CenterSophia, Rotterdam, Netherlands, 3Department of Cardiology, Erasmus Medical Center, Rotterdam, Netherlands, 4Department of Pathology, Erasmus Medical Center, Rotterdam, Netherlands.

Inherited vascular anomalies are commonly characterized by autosomal dominant inheritance with high penetrance, highly variable expressivity regarding size, number and localization of malformations, and small size of postnatal lesions. In 1994, when we identified the genomic locus linked to inherited cutaneomucosal venous malformations (VMCM), we hypothesized that the Knudsons double-hit model for retinoblastoma could be applicable to inherited vascular anomalies. Rarity of accessible resected inherited vascular malformations has hindered such studies. In 2002, we reported a somatic second-hit in one glomuvenous malformation (GVM) and in 2009 another one in VMCM. In this study, we screened 16 GVM tissues for somatic 2nd-hit mutations using DNA extracted from whole tissue or laser capture microdissected tissue, cDNA made of total tissular RNA, or whole tissular DNA for DNA microarray-based analyses. We identified a somatic 2nd-hit in 11/16 lesions (65%). Three were intragenic changes leading to altered mRNA splicing, one was a 1p21-22 deletion and seven were acquired uniparental isodisomies (aUPID) of the whole short arm of chromosome 1. Difficulty of identification, enrichment of somatic mutations by LCM and cDNA studies, and the need for pairwise copy number analysis suggests important tissular heterogeneity for the 2nd-hits. The cellular double-hits lead to localized complete glomulin loss-of-function. Thus, the inherited mutations are phenotypically recessive and need a coexisting somatic mutation, a hallmark of paradominant mode of inheritance. The data demonstrate, for the first time, that aUPID is involved in non-malignant disorders and although the 1p aUPID was specific to

Background: Thoracic aortic aneurysms (TAA) are familial in at least 15-20% of the cases and can be classified in syndromic and non-syndromic forms. The TGF-beta signalling pathway plays a central role in the pathogenesis of both syndromic and non-syndromic TAA. Methods and results: We present a large four-generation family with a syndromic form of TAA. Thirty family members had an extensive physical and cardiologic examination. Eleven familymembers were considered affected on the basis of cardiac and/or skeletal and connective tissue abnormalities. Nine patients had an aortic or other large artery aneurysm. TAA patients had a high risk of aortic dissection or rupture at an early age. Many TAA patients had additional heart malformations, including mitral valve abnormalities, persistent ductus arteriosus and pulmonary valve stenosis. Histological examination of the aorta showed medial fragmentation, disarray of elastic fibres and excess of collagen deposition. After excluding mutations in the known syndromic TAA genes (FBN1, TGFBR1, TGFBR2, COL3A1), we performed a genome wide linkage analysis in this family using the Affymetrix 250K Nsp arrays. A new locus on chromosome 15q with a significant LOD score of 3.6 was identified within a critical region containing 120 genes. Sequencing of positional candidate genes is ongoing. Conclusions: The clinical phenotype overlaps with known TAA syndromes such as Marfan syndrome, Loeys-Dietz syndrome and vascular type Ehlers-Danlos syndrome. Our data provide evidence for a new locus for a syndromic form of TAA. Identification of this novel gene will provide insight into the pathogenesis of arterial aneurysms.

Concurrent Sessions
c16.5 Loss of function ENPP1 mutations cause both generalized arterial calcification of infancy and autosomal recessive hypophosphatemic rickets

2
NHS Trust, Harrow, United Kingdom, 2DNA laboratory, Department of Clinical Genetics, AMC University Hospital, Amsterdam, Netherlands, 3Department of Clinical Genetics, Academic Medical Centre, Amsterdam, Netherlands, 4 Wessex Regional Genetics Laboratory, Salisbury District Hospital,, Salisbury, United Kingdom, 5Section of Medical and Molecular Genetics, Department of Paediatrics and Child Health, University of Birmingham, Birmingham Womens Hospital, Birmingham, United Kingdom, 6Wessex Regional Genetics Laboratory, Salisbury District Hospital, Salisbury, United Kingdom, 7Division of Human Genetics, University of Southampton, Southampton, United Kingdom, 8Department of Medical Genetics, University Medical Center, Utrecht, Netherlands, 9Department of Pediatric Genetics, Emma Kinderziekenhuis AMC, Amsterdam, Netherlands.

B. Lorenz-Depiereux1, D. Schnabel2, D. Tiosano3,4, G. Husler5, T. M. Strom1,6; 1 Helmholtz Zentrum Mnchen, German Research Center for Environmental Health, Inst. of Human Genetics, Neuherberg, Germany, 2ChariteUniversittsmedizin Berlin, Department of Pediatric Endocrinology, Berlin, Germany, 3Meyer Childrens Hospital, Rambam Medical Center, Pediatric Endocrinology, Haifa, Israel, 4Technion-Israel Institute of Technology, Faculty of Medicine, Haifa, Israel, 5Medizinische Universitt Wien, Department of Pediatrics, Vienna, Austria, 6Klinikum rechts der Isar, Technische Universitt Mnchen, Inst. of Human Genetics, Munich, Germany.

The analysis of rare genetic disorders affecting phosphate homeostasis led to the identification of several proteins that are essential for the regulation of phosphate homeostasis, for example fibroblast growth factor 23 (FGF23) a phosphaturic factor, which inhibits phosphate reabsorption and 1,25-dihydroxyvitamin D synthesis in the proximal renal tubules. Cases of hypophosphatemia remain, including familial and consanguineous ones, which do not show mutations in any of the three known genes - PHEX (XLH [MIM 307800]), FGF23 (ADHR [MIM 193100]) and DMP1 (ARHP [MIM 241520]) which, if mutated, causes hypophosphatemic rickets. Here, we present three different homozygous, presumable loss of function mutations in a further gene, ENPP1 (ectonucleotide pyrophosphatase/phosphodiesterase), in members of four families affected with hypophosphatemic rickets. Intact plasma levels of FGF23 were clearly elevated in one of five affected individuals, providing a possible explanation for the phosphaturia and inappropriately normal 1,25-dihydroxyvitamin D levels. Surprisingly, ENPP1 loss of function mutations have previously been described in generalized arterial calcification of infancy (GACI [MIM 208000]), a severe autosomal recessive disorder with a hypermineralizing phenotyp, suggesting an as yet elusive mechanism which balances arterial calcification with bone mineralization. With the identification of ENPP1 mutations as the cause of hypophosphatemia, we added a further component to the growing list of genes involved in the regulation of phosphate hoemostasis. c16.6 Epigenotype-phenotype correlations in silver-Russell syndrome

Silver-Russell syndrome (SRS) is characterised by intrauterine growth restriction, poor postnatal growth, relative macrocephaly, triangular face and asymmetry. Maternal uniparental disomy (mUPD) of chromosome 7 and, hypomethylation of the imprinting control region (ICR)1 on chromosome 11p15 are found in around 5-10% and up to 60% of patients with SRS, respectively. As many clinical features of SRS are non-specific and may change with time, the diagnosis remains difficult. Studies of patients in whom the molecular diagnosis is confirmed therefore provide valuable clinical information regarding the condition. We undertook a detailed, prospective study of 64 patients with mUPD7 (n=20) or ICR1 hypomethylation (n=44). The considerable overlap in clinical phenotype makes it difficult to distinguish these two molecular subgroups reliably. ICR1 hypomethylation patients were more likely to be scored as classical SRS. Asymmetry, fifth finger clinodactyly and congenital anomalies were more commonly seen with ICR1 hypomethylation, whereas learning difficulties and referral for speech therapy were more likely with mUPD7. No correlation was found between clinical severity and level of ICR1 hypomethylation. Use of assisted reproductive technology in association with ICR1 hypomethylation appeared increased compared with the general population. ICR1 hypomethylation was also observed in siblings, though recurrence risk remains low in the majority of cases. Overall, a wide range in severity was observed, particularly with ICR1 hypomethylation. No clinical feature was present in all cases and even low birth weight only in 78%. We would therefore recommend a low threshold for investigation of patients with features suggestive, but not typical, of SRS.

E. L. Wakeling1, M. Alders2, J. Bliek3, H. Bullman4, D. H. Lim5, D. J. Mackay6,7, M. M. Van Haelst8, I. K. Temple6,7, J. M. Cobben9; 1 North West Thames Regional Genetic Service, North West London Hospitals

Genetic counseling Genetics education, Genetic services, and Public policy Abstracts of EsHG Posters P01 Genetic counseling, including Psychosocial aspects, Genetics education, Genetic services, and Public policy
P01.01 misunderstanding and misuse of Genetics counseling at the Expenses of medical Genetics Rules and Basic Human Rights
M. Shariaty, A. Daneil; Tehran Uni.Cancer Research Institute, Tehran, Islamic Republic of Iran.

propose a solution that is both respectful of the childs preferences and that poses not too much administrative overhead for the researchers. Third, we discuss the issue of returning individual research results and incidental research findings for young participants. We argue that such results, when minors are concerned, deserve special attention in guidelines and recommendations. P01.03 the scope of prenatal diagnostics: broad or narrow? Ethical reflections.

Genetic counseling began with the Eugenics movement in Europe and USA around 1910 leading to very destructive inhuman negative effects on health and dignity of humankind. Fortunately the misuse of Eugenic was officially abandoned in 1944. Though it is persists in many developed and developing countries up to today in a rather hidden manner. Clinical genetic counseling based on DIGNITY and EQUALITY of mankind was introduced by Reed in 1944. In Iran and the Islamic countries Genetic counseling was introduced in 1968 by the first author on the same basic principles taught by his graduate mentor professors Yamada, Fuhrmann, Vogel and Reed. These Principles give consideration to ALL of the Humanitarian, Ethical, Moral, Religious and Medical issues concerning the clients and their society On these principles, since 1968 in Iran the first author have provided a genetic counseling service to over 18500 couples seeking academic advice concerning consanguineous marriage, their risk assessments and promer medical advices. Since 1980 due to higher demand for counseling services some unauthorized and un- or inadequately educated MD or PhD holders have started genetic counseling with improper advices or test recommendations in many occasions. These inappropriate or misused issues are NO Marriage, NO Child for many of carriers of monogenic diseases and clinically unjustified Karyotyping of quite normal and healthy young couples and forecasting all of the genetics abnormalities of conception from couples karyotypes .This last issue unfortunately is taken by the lay people as a MUST test for avoiding genetic diseases in their offspring. False positive and negative hopes as well as spending a very time consuming, expensive and inappropriate practice is among these malpractices of genetic counseling in Iran and many developing countries. We feel it is time for WHO and international medical genetic congresses to address this very humanitarian and academic issue and draw a very precise nomenclature for the practice of Genetic Counseling all over the world. Also the very scientific fact of presence of a general 57% chance of having an abnormal child for every healthy, normal and unrelated young couple without any clinical indications of a genetic abnormality is a fix and fusible fact cannot be avoided through couple testing. This is the price of nature for unknown biological task of continuing generations of mankind. P01.02** the use of stored tissue samples from children for genetic research. Ethical issues.
K. Hens, K. Dierickx; Centre for Biomedical Ethics and Law, Leuven, Belgium.

A. de Jong1,2, W. J. Dondorp1,3, G. M. W. R. de Wert1,3; 1 Maastricht University, Research Institute GROW, Maastricht, Netherlands, 2 Centre for Society and Genomics, Nijmegen, Netherlands, 3Maastricht University, Research Institute CAPHRI, Maastricht, Netherlands.

Various new techniques of rapid aneuploidy diagnosis (RAD) challenge the tenability of karyotyping as the gold standard of prenatal diagnosis for pregnancies at increased risk. RAD, targeted at a few chromosomal abnormalities, generates less so-called unexpected findings. This would have the advantage of simplifying counselling, alleviating the burden of choice and preventing unnecessary abortions. Furthermore, RAD would allow mending the present rift between a narrow entrance gate to prenatal screening (risk-assessment for aneuploidy of chromosomes 21, 18, 13) and the much wider scope of subsequent karyotyping. However, RAD misses some clinically relevant abnormalities and may therefore be seen as making suboptimal use of foetal material obtained through invasive and risky procedures. Moreover, it can be asked whether the advantages of having rapid and unequivocal results justify depriving pregnant women of potential useful information. Various views on possible aims of prenatal screening are relevant to this debate, although not conclusive for determining the precise scope of prenatal testing. One possible option may be to provide women with a choice between karyotyping and RAD. The opposing perspectives are also informed by various views on the principle of autonomy: should the emphasis be on optimizing the process of decision-making or on maximising reproductive options? The latter view may imply welcoming the use of tests with an even broader scope than karyotyping. Our preliminary moral assessment suggests that offering RAD as a stand alone test would unjustifiable trespass on reproductive autonomy and amount to a rather restricted view on the aim of prenatal screening. P01.04 EPmA: Predictive, Preventive & Personalised medicine as a novel strategic trend in Europe

D. Radojkovic1, O. Golubnitschaja2, V. Costigliola3; 1 IMGGE, Belgrade, Serbia, 2Department of Radiology,University of Bonn, Bonn, Germany, 3European Medical Association, Brussels, Belgium.

Collections of biological samples have a great potential for genetic research. Much has been written in the last decade on the ethical aspects of the use of such samples from adults, but the ethical issues surrounding the use of stored tissue samples from minors continue to cause controversy. This was made clear this summer by the publication of a policy forum in Science. During the last three years we have conducted theoretical and empirical research on this topic, and will present the conclusions of this research in this talk. We discovered three main areas of interest. First, the requirement that research on vulnerable populations such as children should pose no more than minimal risk is problematic for biobank research, as it is as of yet unclear what risk is entailed in such research. We propose an approach of burden and benefit rather than one of minimal risk to decide on the conditions for participation of children. Second, we reflected on the scope of parental consent. Should parents be allowed to give broad consent for any type of research and for an unlimited amount of time? If and when should the children themselves be involved? We

Predictive diagnostics is considered as the basis for targeted preventive measures and consequent development of individualized treatment approaches. Of paramount importance is the communication among professionals - medical doctors, biotechnologists, computer-scientists, healthcare providers, policy-makers, educators, who are obligatorily involved in the paradigm change from curative to predictive medicine. The paradigm change can be achieved only by well-coordinated action towards : adequate investment in creating novel technologies, development of non- or minimally-invasive diagnostic tools, well-organised exchange and transfer of knowledge among biomedical research entities and biotech industries for production of the advanced diagnostic tools, quality assurance through the introduction of international standards for technological tools and devices, patenting and licenses, professional education in terms of the application of biotechnological high-tech in medicine, political regulations in the health-care sector: introduction of the obligatory guidelines and clear regulations for the health insurance industry to ensure patients needs are met, measures to ensure confidentiality of patient information and personal databank and distribution of relevant information among health-care professionals and users. The mission of the European coordinator in this field is performed by the European Association for Predictive, Preventive and Personalised Medicine (EPmA). The Association is clearly structured in order to reach the possibly best coordination of the PPPM-related multifaceted activities over the whole Europe: there are National Representatives in all 27 Country-members of the European Union and the Associated-Countries (e.g. Israel, Serbia, etc).

Genetic counseling Genetics education, Genetic services, and Public policy

P01.05 timE-trial: A multicentre study on the behavioural and psychosocial effects of rapid genetic counselling and testing in newly diagnosed breast cancer patients P01.07 Cardiac genetics: the first 250 clinic patients

M. R. Wevers1,2, M. G. E. M. Ausems2, E. M. A. Bleiker1, D. E. E. Hahn1, T. Brouwer2, E. T. J. Rutgers1, R. van Hillegersberg2, F. Hogervorst1, R. B. van der Luijt2, H. Valdimarsdottir3, S. Verhoef1, N. K. Aaronson1; 1 Netherlands Cancer Institute-Antoni van Leeuwenhoek Hospital, Amsterdam, Netherlands, 2University Medical Centre Utrecht, Utrecht, Netherlands, 3Mount Sinai School of Medicine, New York, NY, United States.

J. McGaughran1, J. Atherton2; 1 Genetic Health Queensland, Brisbane, Australia, 2Department of Cardiology, Royal Brisbane and Womens Hospital, Brisbane, Australia.

Background: In 15% of high-risk families, a BRCA1or BRCA2 gene mutation can be found. Female carriers with breast cancer have an increased risk of new primary tumours and may opt for preventive surgery. Usually, genetic counselling and DNA-testing are offered after primary treatment and take 4-6 months. However, some Dutch laboratories can generate test results within 3-6 weeks. Little is known about the effect of such rapid genetic counselling and testing (RGCT) procedures on treatment decisions and psychosocial health. Methods: The TIME-trial is a prospective randomized study. Newly diagnosed breast cancer patients (N=255) with 10% or greater chance of having a BRCA mutation are being recruited from 12 Dutch hospitals. They are randomized on a 2:1 basis to either a RGCT-group (referral to a clinical geneticist within one week of diagnosis), or a usual care control-group. If needed, DNA-test results are made available within 3-6 weeks. The primary study outcome is choice of surgical treatment. Secondary outcomes include perceived cancer risk, cancer-related distress, health-related quality of life and decisional satisfaction. Psychosocial assessments take place at study entry and at 6 and 12 months follow-up. Results: Patient recruitment started in November, 2008. To date, 124 women (response 82%) have been entered into the trial. Of the 80 participants in the RGCT-arm, approximately one-third has opted for rapid DNA-testing. Conclusion: This multicenter clinical trial of RGCT is recruiting well and, based on the figures available to date, a higher percentage of women is opting for rapid genetic testing than had been anticipated. P01.06 introducing a patients choice in how to learn the results of presymptomatic DNA testing for hereditary cancer
J. C. Oosterwijk, J. S. Voorwinden, J. ter Beest, B. K. Leegte, R. H. Sijmons, J. P. C. Jaspers; UMCG, Groningen, Netherlands.

In February 2007, a monthly cardiac genetics clinic was commenced at the Royal Brisbane and Womens Hospital. The clinic was to ensure appropriate counselling, genetic testing and management of those with or at risk of an inherited cardiac condition. A range of conditions have been seen including; hypertrophic cardiomyopathy, familial dilated cardiomyopathy, arrythmogenic right ventricular cardiomyopathy, catecholaminergic polymorphic ventricular tachycardia and long QT syndrome. Rarer conditions such a Fabry disease and Danon disease have also been seen. To date 250 patients have been seen through the clinic. Because of pressure on clinic space, a number of the patients referred to the genetic service have been seen in genetics clinics at locations around the state. A number of challenges have been encountered: management of those with a mutation but no phenotype; testing of at risk children; funding for testing; waiting times with increasing numbers of cases referred. The clinic also collaborates with the forensic pathology service and coroner in the assessment of those families where there has been a sudden unexplained death in a young individual. The data from the clinic will be presented P01.08 Ethical aspects of Array Comparative Genomic Hybridisation (cGH) for mental retardation diagnosis and genetic counselling

S. JULIA1,2, A. Soulier2, D. Sanlaville3,4, E. Rial-Sebbag2, B. Keren5, D. Heron5, M. Till3, P. Edery3,4, P. Calvas1, G. Bourouillou1, A. Cambon-Thomsen2; 1 Service de Gntique Mdicale,, toulouse, France, 2Inserm, U558, Epidemiology and analyses in public health and University Toulouse III, Toulouse, France, 3Service de Cytogntique Constitutionnelle, CHU de Lyon, Bron, France, 4EA 4171, Universit Claude Bernard, Lyon 1, Lyon, France, 5 Dpartement de gntique, Hpital de la Piti Salpetrire, Paris, France.

AIM: To test whether coping after DNA testing for hereditary cancer, is similar when counselees have the choice to learn the results by letter instead of in a face to face contact. BACKGROUND: In pre-symptomatic testing for hereditary cancers, the availability of treatment /prevention seems to mitigate the impact of the test result, as compared to Huntingtons chorea. Moreover, counselees indicated they preferred to learn the test results by letter for several reasons. METHODS: Cases referred for pre-symptomatic testing for BRCA or Lynch syndrome were randomised between standard care protocol (disclosure of DNA results in a face to face contact), and choice protocol (the counselee gets the choice to either learn the result in a face to face contact or to be sent the results in a letter with a subsequent appointment). Included counselees filled out questionnaires covering knowledge, coping, stress and satisfaction before the intake session (t1), 2 days after DNA test result disclosure (t2) and 6 weeks later (t3). RESULTS: In total 244 out of 340 ascertained cases consented in the study and returned the first questionnaire: 205 BRCA1/2 and 39 Lynch syndrome. Some refrained from or postponed DNA testing after intake and were excluded To date 48 cases got the standard protocol (20 carriers, 28 non-carriers) and 82 cases got the choice protocol (26 carriers and 56 non-carriers). Of the 82 cases in the choice protocol, 58 chose disclosure by letter and 24 chose disclosure in an appointment. The results of our analysis will be presented.

The recent rapid emergence of new technologies in genetic medicine contributes to the diagnosis and the understanding of the molecular basis of numerous diseases. The high throughput and the sensitivity of these technologies will create an unprecedented volume of information for patients, counsellors, and health care providers, raising also unique ethical challenges. Although the applications of many of these techniques are still limited to research, Array Comparative Genomic Hybridisation (CGH) has evolved to a standard application in clinical genetics, especially in individuals with syndromic or non-syndromic mental retardation. CGH allows a whole genome analysis at a resolution, 10-10 000 times higher than that of routine chromosome analysis by karyotyping. This method has revolutionized the present clinical practice in the detection and diagnosis of human chromosome abnormalities in mental retardation. Some ethical issues specifically challenge the transfer of these improved technologies from the research laboratory to their clinical applications. In our experience we have identified several situations asking ethical difficulties. They will be presented according to two main dimensions: uncertainty of results and incidental findings. Ethical issues of genetic disease diagnostics and various health applications have already been addressed in numerous reports, but with the use of these screening technologies in the clinical setting, the incidental becomes usual, representing new challenges for the clinician. Specific reference documents addressing the ethical issues of genetic testing in relation to CGH are still lacking. P01.09 Evidence-based information guides to rare chromosome disorders for families and professionals
B. A. Searle1, P. Middlemiss1, S. Wynn1, M. Hulten2; 1 Unique, Caterham, United Kingdom, 2University of Warwick, Warwick, United Kingdom.

Purpose: To develop reliable, relevant, accurate leaflets for affected families and health professionals that fill an information gap about rare chromosome disorders . Method: In 2003 Unique surveyed information materials published in the UK about specific rare chromosome disorders: for over 93% of members no accessible disorder-specific information was available. Unique asked families what they most wanted to know at diagnosis

Genetic counseling Genetics education, Genetic services, and Public policy

and what questions remained unanswered. Unique prioritised 66 disorders according to frequency on its database (7122 member families at February 2010) and absence of existing information accessible to families. Information was compiled from the medical literature; from Uniques database; and from detailed surveys sent to member families. Draft texts were reviewed for accuracy by Uniques medical adviser and by medical and genetics professionals expert in the specific disorders. Photographically illustrated draft leaflets were vetted for content by families. Results: Leaflets have been developed on 113 topics including numerical and structural disorders; subtelomere deletions; mosaic disorders; emerging microdeletion and microduplication syndromes and a broad range of less common diagnoses. Discussion: Twenty-one leaflets have been translated into at least one European language. Leaflets, available to families and health professionals in print format and online at www. rarechromo.org, improve families understanding and acceptance of a rare chromosome disorder and help diminish the acute stress and anxiety associated with diagnosis. P01.10 teaching the genetics of complex traits

type C/C in the group of environmental risk (68.2% versus 59.87% in the comparison group, P = 0.0478). For a comprehensive analysis was studied genotypes of the five polymorphism in the studied genes. Revealed a statistically significant increase of genotypes for the study of polymorphism at environmental risk (AADDAGCCCC: 2 = 9,69, P = 0.0027, AGDDAACCCC: 2 = 9,47, P = 0.003), may determine that a combination of data most efficient biotransformation of xenobiotics and reduces the negative impact of environmental factors and some carcinogens in the body. P01.12 medical and genetic counseling of families with cystic fibrosis in the Republic of Moldova

N. I. Barbova; State Center of Reproductive Health and Medical Genetics, Chisinau, Moldova, Republic of.

M. J. Dougherty; American Society of Human Genetics, Bethesda, MD, United States.

The primary goals of secondary and post secondary science education are to help prepare scientifically literate citizens and future scientists. Efforts to improve science education have focused on competency standards and the professional development of teachers, and recent efforts have promoted the use of high-stakes exams and benchmarks for school performance. In spite of this work, students in the United States continue to lag behind their peers in other countries. This underperformance is true for genetics and for science and math in general, and it is particularly worrisome given the accelerating need for scientists and engineers in an increasingly technology-driven economy. Unfortunately, even students in high-performing European countries, such as Finland and Slovenia, have relatively small percentages of students performing at the higher levels of proficiency. Perhaps nowhere is the need for scientific literacy more personally meaningful and urgent than in genetics--because of its connection with medicine. Rapid changes in human genetics--in particular recent insights into the genetics of complex traits--have the potential to transform healthcare. However, citizens are ill-prepared to participate in this transformation because the curriculum is outdated and remains focused on single-gene inheritance and simplified notions of risk. One potential solution is to modernize the genetics curriculum so that it matches the science of the 21st-century. This paper outlines the problems with current genetics instruction, highlights changes in human genetics that support a curricular reorganization, and proposes a new genetics curriculum for secondary and postsecondary education. P01.11 molecular genetic study of polymorphic variations in genes within the cascade of reactions detoxification of xenobiotics with aromatic structure

Cystic fibrosis is an important medico-social problem for the Republic of Moldova, which is associated with low life expectancy of patients (average age - 13,5 years), the difficulties of clinical and molecular diagnostics, low economic level of the population and lack of social support for families with cystic fibrosis. An important step in the prevention of cystic fibrosis is a medico-genetic counseling in families at high risk. We are presenting an 18-year experience of retrospective counseling of 58 families in which there were 69 children suffering from cystic fibrosis aged from 1 month to 18 years (35 boys and 34 girls). In 56 families there were moral and ethical difficulties with improvement of the diagnosis. Most families (63.8%) lived in rural areas, 66,4% of parents had secondary education. The survey showed that 72% of parents knew that the disease has a genetic nature. The results of DNA testing showed mutations in both alleles in 14 families, mutation of one allele in 25 families and 19 families were non-informative for molecular diagnosis. The prenatal diagnosis carried out in 6 families resulted to 4 healthy children. Of the 58 families observed in 52 families there are no children with CF, that is, the effectiveness of genetic counseling was 89.7%. Information of parents about the presence of high risk have genetic nature of CF lead to positive changes of reproductive behavior. The prenatal diagnosis in affected families can allow decrease the frequency of ill children. P01.13 the opinion of cF patients parents about different aspects of genetic services in Russia

L. Y. Ivanova1, V. L. Izhevskaya2, I. V. Zhuravleva1, E. K. Ginter2; 1 Institute of Sociology of Russian Academy of Sciences, Moscow, Russian Federation, 2Research Centre for Medical Genetics of RAMS, Moscow, Russian Federation.

E. M. Vasilyeva, E. V. Vorobjeva, V. Y. Gorbunova; Bashkir State Pedagogical University of the name M. Akmulla, Ufa, Russian Federation.

Development of industrial production around the world have led to a large number of chemical compounds that adversely affect human health. Toxic substances violate the metabolism, the structure of cells and tissues, resulting in abnormal reaction of the organism. The negative effect of xenobiotics depends on the activity of physical and biochemical methods of protection, including the immune system and the system of biotransformation of alien connections. We studied DNA samples - 714 apparently healthy individuals in the two groups, differing in environmental conditions of residence in the same territory, examined polymorphism of genes within the cascade of reactions detoxification of xenobiotics with aromatic structure: CYP1A1/A4889G, CYP2E1/Ins 96,EPHX1/A4156,NQO1/C609T,C465T. A group of environmental risk revealed a significant increase in the frequency of the protective genotype D / D polymorphism Ins96 gene CYP2E1 (79,74% against 58.44%). Comparative analysis of the frequency distribution of genotypes and alleles of polymorphism C609T gene NQO1 revealed a statistically significant difference by increasing the frequency of the protective geno-

The newborn screening for CF started in Russia in 2006. The opinion of CF patients parents to different aspects of genetic services in Russia was estimated. 68 respondents were participated in the research. Majority of parents (90 %) have learnt about hereditary character of their child disease from the pediatrician, only 81 % of them have been referred to genetisist, and 67 % have been held DNA testing. However 90% respondents have consider, that they have understood the information about repeated genetic risk for CF. However only 50% of them could correctly specify the value of recurrence risk of CF, and only 30 % of them could correctly attribute a risk category. The birth of the CF child has affected reproductive plans in 30 % of CF families, only 35% of families wanted to have more children. 80 % of CF childrens parents knew about possibility of prenatal diagnostics of CF, however almost 40 % from them have learnt about it not from experts, but from the press. Prenatal diagnostics of CF was acceptable for 66% of respondents, and the abortion of a CF foetus is morally acceptable for 82% of them. Only 4 % of respondents have answered that they didnt want to terminate the CF foetus pregnancy. In 11/15 women who were pregnant after the birth of CF child, prenatal diagnostics of CF was done: 7 foetuses were healthy, 4 - with CF. All CF foetus pregnancies have been terminated. P01.14 Psychosocial aspects of genetic paternity testing ordered by courts: does biological relatedness create a father?
H. Machado1, S. Silva2, S. Costa3, D. Miranda3, A. Amorim4, C. Alves5; 1 Department of Sociology, University of Minho and Center for Social Studies, University of Coimbra, Portugal, 2Institute of Public Health (ISPUP),

Genetic counseling Genetics education, Genetic services, and Public policy

Department of Hygiene and Epidemiology, University of Porto Medical School, Portugal, 3Center for Social Studies, Coimbra, Portugal, 4Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP) and Faculty of Sciences, University of Porto, Portugal, 5Parentage testing and Genetic Identification Unit, Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP), Portugal.

Colentina Hospital, Bucharest, Romania, 5Psychiatric Hospital, Poiana Mare, Romania, 6Romanian Parent Project Association, Craiova, Romania.

Many European societies support efforts to establish genetic parentage of children born outside marriage when no father is listed on the birth registration. This State effort is usually part of public policies to ensure that children are cared for not only financially but also with regards to education and upbringing. There are also medical reasons to establish genetic parentage, and the importance of parents in the psychological development of infants. We discuss the benefits and/or disadvantages of genetic paternity testing from the point of view of mothers and fathers. We focus on cases ordered by Portuguese courts of law and rely on data proceeding from interviews conducted with women and men who went through the process of DNA paternity testing. Some topics of debate are: psychosocial aspects of submission to genetic paternity testing; informed consent; access to information about genetic testing procedures and communication of results; perceptions of the role of genetic relatedness in defining fatherhood. The results suggest that there are important psychosocial impacts produced by genetic paternity testing which question the value of the legal primacy of biological relatedness in forming fatherhood and in serving the best interests of the child. We recommend better practices of informed consent, communicating the results of paternity tests and even psychological counseling to motivate mothers and fathers for the necessity of creating and continuing a relationship with the child and between the progenitors. These practices demand a more effective coordination between courts of law and laboratories and specific training of personnel. P01.15 DOUBLE cOUsiN mARRiAGE iN tHE iRANiAN PROViNcE OF HORmOZGAN (2002-2009)

The right management of Duchennes muscular dystrophy (DMD) should include several steps: a) clinical and genetic diagnosis and counseling, b) refer to a specialist and multidisciplinary team, c) information and support for the family. Our study was conducted for 16 Romanian boys affected by DMD and their families. We examined how the diagnosis was made, the application of specific treatment with corticosteroids, tracking of supportive therapy protocol, involvement of a team of specialist in the disease management, genetic counseling and psychosocial support for families. The results of this study revealed a system of management of this disease fractured at all levels: - Late clinical diagnosis (4 cases), late genetic diagnosis (8 cases) - Incomplete diagnosis - lack of immunohistochemical diagnosis and/ or molecular diagnosis (6 cases) - Wrong diagnosis (2 cases, although diagnosed with DMD, after molecular analysis it is being evaluated for a different type of dystrophy). - Lack of therapeutic instrument provided by a multidisciplinary team of specialists, inadequate treatment (15 cases) - Lack of genetic counseling (5 cases) and psychosocial support to families (15 cases) From the 16 children, only one benefits by a correctly and completely mangement of the disease. In the context of the emergence of the first innovative treatment (PTC 124, exon-skipping), if we want Romanian children to have access to clinical trials, it is vital to have a standard of care compatible with the European one; the management of DMD by a network of specialists can buy time for our children and stop the reappearance of the disease. P01.17 Population-based study of dystrophin mutations in canada

P. Nikuie, G. Tabasi, F. Hajizadeh; Bandarabbas soicial welfare organization, Bandarabbas, Islamic Republic of Iran.

Background: Double first cousins arise when siblings of one family reproduce with siblings of another family. The resulting children are related to each other through both parents families. Double first cousins share both sets of grandparents in common and have double the degreeof consanguinity than ordinary first cousins.Genetically, they are as related as half -sibling sharing 25 %of their DNA. A consanguineous couple especially double cousin couple is at increased risk for both autosomal recessive disorders and several congenital malformations. Material and methods: In seven years from 2002 to 2009,we performed genetic counseling for 2400 couples.The consanguinity between 34 couples was double cousin.(1.4%)Familial pedigree established and risk of genetic disorders for their children was calculated. Results: 1428 couples (59.5%) had consanguinity and 34 couples (1.4%) were double cousins. -13 couples (38.3%) came premariage, 17 couples preconception (50%) and 4 couples (11.7%) during pregnancy. -inbreeding coefficient in 15 couples (44.1%) was more than 1/8. -18 couples (52.9%) had positive history of genetic disorders in family. -15couples had children and in 14 of them (93.3%) at least one child was involved with genetic disorders. -3 couples (8.8%) were carrier of beta thalasemia and referred for prenatal diagnosis. Conclusion: Based on high risks for genetic disorders in double cousin marriage and according to all of couples announced if they knew about these risks, they would avoid marriage. Performing genetic counseling is recommended before all consanguious marriages especially double cousin type. P01.16 Management deficiencies of Duchenne Muscular Dystrophy: A reality that we can changes

G. Yoon1, J. K. Mah2, K. Selby3, C. Campbell4, A. Nadeau5, M. Tarnopolsky6, A. McCormick7, J. Dooley8, H. Kolski9, A. Skalsky10, G. Smith11, D. Buckley12, P. Bridge2, P. N. Ray1; 1 The Hospital for Sick Children, University of Toronto, Toronto, ON, Canada, 2 University of Calgary, Calgary, AB, Canada, 3University of British Columbia, Vancouver, BC, Canada, 4University of Western Ontario, London, ON, Canada, 5 Universite de Montreal, Montreal, QC, Canada, 6McMaster University, Hamilton, ON, Canada, 7University of Ottawa, Ottawa, ON, Canada, 8Dalhousie University, Halifax, NS, Canada, 9University of Alberta, Edmonton, AB, Canada, 10 University of Manitoba, Winnipeg, MB, Canada, 11Queens University, Kingston, ON, Canada, 12Memorial University, St. Johns, NL, Canada.

A. Dobrescu1, F. Leturcq2, M. Ioana3, M. Alexianu4, L. Sirbu5, I. Tudorache6; 1 Department of Medical Genetics, University of Medicine and Pharmacy, Craiova, Romania, 2Biochemistry and Molecular Genetics Department, Cochin Institut, Paris, France, 3Department of Cell and Molecular Biology, University of Medicine and Pharmacy, Craiova, Romania, 4Department of Neuropathology,

Background: Duchenne and Becker muscular dystrophy (DBMD) are allelic disorders caused by mutations of the dystrophin gene on Xp21. This study describes the diagnostic methods and mutation frequency among the patients of the Canadian Pediatric Neuromuscular Group (CPNG). Methods: De-identified data containing the clinical phenotypes, diagnostic methods, and mutational reports from DBMD patients followed by CPNG centers during 2000-2009 were analyzed using descriptive statistics. Patients were considered to have had complete genetic testing if all 79 exons of the dystrophin gene were examined and if this was negative, sequencing of the gene. Diagnosis solely by muscle biopsy or PCR analysis of a subset of exons was considered incomplete genetic testing. Results: 773 DBMD patients had a confirmed diagnosis based on genetic testing (97%), muscle biopsy (2.3%), or family history (0.7%). 573/773 (74%) had complete deletion/duplication analysis of all 79 exons or gene sequencing resulting in 366 (64%) deletions, 64 (11%) duplications, and 143 (25%) point mutations. The most common point mutations were nonsense (47%), followed by frameshift (31%), splicing (15%), and missense/amino acid deletions (7%). Access to complete genetic testing was variable and ranged from 88% for patients from ON to 35% for patients from NS. Conclusion: This is the first comprehensive report of dystrophin mutations in Canada. It highlights the need for complete genetic testing of patients with dystrophinopathy, and the necessity for collaboration among academic centers and neuromuscular disease registries to ensure that patients are receiving optimal care and are eligible for mutation-specific therapies.

Genetic counseling Genetics education, Genetic services, and Public policy

P01.18 Influencing how genetics is taught in UK secondary schools: the Nowgen schools Genomics Programme

L. E. Holmes1,2, P. Finegold2, I. Starling1,2, K. Dack1,2, A. Hall3, J. Worthington2, J. Harris2, A. Read2, H. R. Middleton-Price1,2, D. Donnai1,2; 1 Nowgen, A Centre for Genetics in Healthcare, Manchester, United Kingdom, 2 Manchester Academic Health Science Centre, The University of Manchester, Manchester, United Kingdom, 3Nuffield Foundation, London, United Kingdom.

Current research in human genomics has great potential for sparking interest amongst secondary school students, yet it is not prominent in UK school curricula. The Nowgen Schools Genomics Programme aims to redress that situation; narrowing the gap between genomics research and classroom genetics. The genetics content of UK school curricula currently concentrates on single-gene genetics with few references to more recent approaches exploring the human genome. Funded by a three-year grant from The Wellcome Trust, the Nowgen Schools Genomics Programme brings together leading scientists, clinicians, educationalists and bioethicists to contribute to a range of approaches designed to equip young people to assess the real potential of genomics, and to make informed decisions about future healthcare. The programme will introduce genomewide association studies, pharmacogenetics and genetic medicine to teachers and their students, and will support them to: - explore genetic and lifestyle contributions to disease; - consider the methodological challenges contemporary large population studies require; - examine the social and ethical challenges to society; - explore the potential impact of data arising from contemporary genetics. P01.19 DNA Day:a good starting point to disseminate genetic education in high school

disclosure of individual genetic data to research participants is ethically imperative. On the other end of the spectrum, it is argued that no individual genetic research results should be disclosed whatsoever. An intermediate position is defended by those who opt for a qualified disclosure. Although most commentators adopted a variant of this latter position, their underlying argumentation and the conditions for disclosure varies widely. Some for example argue that individual genetic data should only be disclosed when these data are of analytic validity and clinical relevance. Others argue that the decisive factor concerns the relationship between participant and researcher; for example, the more intense this relationship, the stronger the obligation to disclose would be. Clarity on the appropriate standard of disclosure is important in view of the changing genetic landscape, such as the current genome wide association studies, the impending whole genome sequencing-studies and the upcoming commercial activities. In this paper, we present the results of our systematic search and explore whether and when individual genetic data should be returned to research participants. P01.21 construction of an ethics policy for a bioinformatics European project: GEN2PHEN

A. Cambon-Thomsen1, A. Pigeon1,2, E. Rial-Sebbag1, E. U. GEN2PHEN Consortium3; 1 Inserm, U558 & University of Toulouse, Toulouse, France, 2Centre de droit des affaires, Universit Toulouse 1 Capitole, Toulouse, France, 3University of Leicester, Leicester, United Kingdom.

B. Zanini1, M. Borriello2,3, V. Marini4, R. Ravazzolo1; 1 University of Genoa,Istituto G.Gaslini, Genova, Italy, 2University of Naples,Federico II, Naples, Italy, 3Istituto Marco Polo, Genova, Italy, 4University of Genoa, Genova, Italy.

DNA Day-a special day-initiated in 2003 to commemorate the completion of the Human Genome Project in April of that year, and the discovery of the DNA double helix fifty years earlier! In 2008 The European Society of Human Genetics (ESHG) joined the American Society of Human Genetics (ASHG) in making a Europewide celebration of genetics and its promises and decided to expand a very successful initiative: the DNA Day Essay contest. That contest is one of several ongoing initiatives to promote knowledge and understanding of genetics in secondary schools. In Italy, the Dna Day contest has been a fantastic tool and occasion to propose an advandced knowledge in genetics, increasing need of an accurate education at school. The collaboration and support between teachers, the Italian Society of Human Genetics (SIGU) and the University of Genova has carried on inititives of lectures by geneticists in order to close high school students to research topics, facilitate and stimulate a much more wide reflection on the essays proposed by the contest. In 2008, in Genova 400 people (students and teachers) attended the conference at the Faculty of Medicine and in 2009, more than 250 people were present at the theatre of Advanced Centre of Biotecnology. This year,the SIGU has set up a more strict plan to promote a genetic education project: - addressing a letter to the high school teachers - publishing a banner on the national website - having a national selection and awarding three italian students at the national meeting P01.20 Feedback of individual genetic data to research participants: a qualified disclosure?

Genomic technologies including bioinformatics methods and creation of genomic databases have become increasingly important for the scientific community but have increased faster than its possible framing by an appropriate legislation. However theses activities must be developed with respect to fundamental values such as dignity, privacy, autonomy. The lack of a binding instrument regulating all aspects of these issues and the varied and disparate legal and ethical frameworks has resulted in confusion for professionals working in genomics to construct their own ethics policies. Compliance with the various international, European, and National ethical and legal instruments requires a more considered approach. We analysed how a specific bioinformatic European project, GEN2PHEN, is dealing with this issue in the development of its ethics policy. To provide ethical oversight, analysis, and guidance for GEN2PHEN, we considered texts and existing guidelines reviews, an international consortium questionnaire survey of opinions, discussion in general assembly, consultation with relevant EU and international projects and review of recent literature. The main issues identified were requirements for entering data into the system, managing them over time in federated databases and conditions for their accessibility. A process for checking the computer tools produced for ethics compliance was elaborated. The policy takes into account the differences between various types of data, mandatory items of consent and modalities of their verification, clarification of responsibilities within and outside the project, data and database ownership issues and transparency of the process. It involves a project ethics oversight committee and specifies relations with local research ethics committees. P01.22 Eurogene: a pan-european e-learning service in human genetics
M. Dutto1, Z. Zdrahal2, E. Sklavounou3, M. D. Teare4, F. Clerget-Darpoux5, G. Romeo1; 1 European Genetics Foundation, Bologna, Italy, 2The Open University (Knowledge Media Institute), Milton Keynes, United Kingdom, 3Systran, Paris, France, 4University of Sheffield, Sheffield, United Kingdom, 5Universit ParisSud, Paris, France.

A. L. Bredenoord, J. J. M. van Delden; University Medical Center Utrecht, Julius Center, Utrecht, Netherlands.

Last years, a debate evolved regarding the question whether individual genetic research results should be disclosed to research participants in genetic/genomic research, and if so, which data, and by whom. With genetic research results we refer to a broad category of information, including validated and non-validated, highly and poorly predictive, and more or less probabilistic genetic data generated by a medical scientific study. On the one end of the spectrum, it is argued that full

The primary objective of the Eurogene project is to develop the means for accessing, sharing and reusing high quality educational resources in human genetics with multilingual support. The Eurogene portal offers access to learning resources produced by a network of professionals in the field represented by universities, hospitals and educational centres across Europe. The scope of Eurogene includes statistical, medical and molecular genetics and the content is targeted at people with different levels of previous knowledge ranging from the lay person

Genetic counseling Genetics education, Genetic services, and Public policy

to the expert practitioner. Working towards this goal Eurogene has created a genetic domain ontology and a hierarchy of terms that are first used to annotate and enrich the e-learning resources. This automatic annotation forms structure to support the suite of web based tools developed specifically for providing, finding and translating digital educational content in the field of genetics. Interactive machine translation tools allow approved users to refine, train and hence validate the genetic context multilingual dictionary. The Eurogene portal exploits technology which makes it easy to navigate and explore the available articles, images and videos. Educators can thus move toward blended learning in an interactive and natural way, while students are given the opportunity to finally become actors in the educational process.We will demonstrate the main features of the tools offered by the Portal. This will include viewing recent sample content, illustration of the annotation process and machine translation options, and construction of learning packages. EUROGENE is a 36 month project supported by the ECs e-Content Plus program. P01.23 Detection of FMR1 cGG repeat expansions from genomic DNA and single cells - a direct triplet-primed PcR approach

Cancer Registry is a population-based databank with all individual cancer information recorded since 1955. A database-generated pedigree (DGP) can be made by linking IGCD pedigrees to records in the Cancer Registry using personal identification numbers. This results in a very comprehensive cancer family history, potentially supporting more efficient assessment of the individuals inherited cancer risk. In Iceland, cancer genetic counselling based on DGP has been offered since 2007 and 310 counselees have used the service, the majority (97%) because of a breast cancer family history. A counselees pedigree is constructed as well as a DGP (3-4 generations with 150-1600 individuals on each side). The DGP is not revealed at the personal level to the counselee for privacy reasons. Risk assessment for cancer, including possible indication for molecular testing, is based on both the counselees pedigree and the DGP. We are not aware of this approach resulting in any problems or privacy concerns. The DGP and the counselees pedigree need to be compared including cost efficiency and predictive power. If advantages are demonstrated, other communities could address legal issues and generate their genealogical databases. P01.25 Participation of nurse in genetic counseling; present status in Japan

C. Teo1, C. G. Lee2,3, S. S. Chong1,4; 1 Department of Paediatrics, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore, 2Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore, 3Division of Medical Sciences, National Cancer Center, Singapore, Singapore, 4University Childrens Medical Institute and Department of Laboratory Medicine, National University Hospital, Singapore, Singapore.

S. Izumi1,2, H. Yokoyama3,2, M. Ishii3, M. Mizoguchi3,2, A. Kondo1,2, K. Takahashi1,2, Y. Onuki2,4; 1 Department of Obstetrics and Gynecology, Tokai University School of Medicine, Isehara, Japan, 2Dept. Clinical Genetics, Tokai University Hospital, Isehara, Japan, 3School of Health Sciences, Tokai University, Isehara, Japan, 4 Department of Neurology, Tokai University School of Medicine, Isehara, Japan.

Fragile X syndrome (FXS) is the most common cause of heritable mental retardation. It is mostly caused by the hyperexpansion and hypermethylation of CGG repeat expansions in the 5 untranslated region of FMR1, leading to gene silencing. There are 4 allelic classes of FMR1 CGG repeats: normal (5-44 repeats), gray zone (45-54 repeats), premutation (55-200 repeats), and full mutation (>200 repeats). Full mutation alleles are associated with FXS, while unmethylated premutation alleles have been associated with premature ovarian failure (POF) in females and with late-onset fragile X-associated tremor ataxia syndrome (FXTAS) in males. Molecular diagnosis of FXS usually involves PCR across the CGG repeat to determine repeat length, supplemented by Southern blot analysis for male samples with null amplification or female samples with a single allele size by PCR. We describe a highly specific direct triplet-primed PCR (dTP-PCR) approach to detect FMR1 premutation and full mutation expansions. Unique capillary electropherogram patterns also enable determination of repeat lengths and structures of normal, gray zone and small premutation alleles in all males and some females. Detection sensitivity is at the single cell level, paving the way for direct detection of expanded FMR1 alleles in preimplantation embryos. For more rapid single-step screening, dTPPCR products can be immediately subjected to melting curve analysis in the presence of SYBR Green dye to yield distinct, clearly differentiated melting peaks from normal versus expanded alleles, demonstrating the utility of this homogeneous assay as a rapid first-line screen for FXS, POF and FXTAS. P01.24 Linking a genealogical database and a cancer Registry to generate comprehensive pedigrees for cancer risk assessment.

Objective: Since 2005, Tokai University had started a Master course of genetic nursing in School of Health Science, which is only one course in Japan. To improve this condition, we are continuously emphasizing several merits in participation of genetic nurses in genetic counseling clinics. In this study we tried to elucidate the present status of participation of nurse(s) in clinical genetic counseling in Japan, as well as the expected roles of nurse(s) from other medical professionals in genetic counseling team. Method: Questionnaires were mailed on January 2009 to 129 facilities, which are registered as genetic counseling clinics in Japanese Society of Obstetrician and Gynecologist. We focus in the prenatal diagnosis, which is the most common subject in the a genetic counseling clinic, Results: Answers were retrieved from medical doctor, nurse, and psychologist, where the response rate was highest (36.2%) from doctors. Nurses are incorporated in the team on 37.0% of responded facilities, where their role is guidance on the patients daily activity before and after the amniocentesis. On the other hand, they give crucial information about the procedure in fewer occasions. Medical doctors expect nurses in a future to collect the information for counseling, to make a precise pedigree, and to shape up the clients will. Conclusion: Comparing our previous research result, more nurses participate in genetic counseling than double of those on 2000. However, the distribution imbalance between the facilities becomes more prominent than the past. P01.26 Developments in Genetic counselling: a European Perspective

H. Skirton1, M. Voelckel2,2; 1 Faculty of Health, Plymouth, United Kingdom, 2Hpital dEnfants de la Timone, Marseille, France.

V. Stefansdottir1, O. T. Johannsson1, H. Skirton2, L. Tryggvadottir3, H. Tulinius4, J. J. Jonsson1,5; 1 Landspitali, Reykjavik, Iceland, 2University of Plymouth, Plymouth, United Kingdom, 3Icelandic Cancer Society, Reykjavik, Iceland, 4The Icelandic Genetics committee., Reykjavik, Iceland, 5The Department of Genetics and Molecular Biology, Reykjavik, Iceland.

Genetic cancer counselling is generally based the counselees pedigree, i.e. information provided by the counselee and confirmed, when possible through hospital records and occasionally a cancer registry. Electronic genealogical databases allow quick construction of an extensive pedigree, encompassing individuals beyond the counselees knowledge. The Icelandic Genetics Committee Database (IGCD) holds accurate genealogical information based on official demographic records about individuals living in Iceland since 1850. The Icelandic

The European Network of Genetic Nurses and Counsellors was formed in 2008 and now exists under the auspices of the Ad Hoc Genetic Nurse and Counsellor Accreditation Committee of the European Society of Human Genetics. Currently there are almost a hundred members. There is wide variation in the profession across Europe, with many registered practitioners in some countries, and few or none in others. In the interests of patient safety, common minimum standards are required. The aims of the network are to ensure that professionals using the title genetic counsellor are competent to practice. Through a series of workshops and consultation with the membership, guidance for the future of the profession has been agreed. The role of the genetic counsellor includes: identifying the needs of the family, collecting and interpreting genetic information relevant to genetic counselling, making a genetic risk assessment and communicating information about the risk and options available to the family. It

Genetic counseling Genetics education, Genetic services, and Public policy

is envisaged that by 2020 the standard training and education for a genetic counsellor will be the Master degree in genetic counselling. The content of these educational and training programmes must include: human genetics, medical genetics, counselling skills and clinical experience. Genetic counsellors will be expected to practice according to the Code of Professional Practice for genetic counsellors in Europe. Further details will be presented and work is now required to consult widely with those from other professional disciplines to ensure that best practice in genetic counselling is adopted in Europe. P01.27 Ethical Aspects of Genetic testing in Russia

P01.29 MPAG - first steps of a competences-centred programme for genetic counsellors in Portugal
M. Paneque, J. Sequeiros; IBMC, ICBAS - Universidade do Porto, Porto, Portugal.

V. L. Izhevskaya, V. I. Ivanov; Research Centre for Medical Genetics of RAMS, Moscow, Russian Federation.

Studies of ethical problems concerning the application of genomic technologies in public health care in Russia commenced in the late 1980s in connection with the works according to the Russian programme Human Genome. These studies showed that the majority of Russian medical geneticists were inclined to be too directive, especially in cases of genetic counselling concerning prenatal detection of foetal pathology. A substantial part of the specialists (approximately 10 - 20 %) were inclined to in some way or another exert pressure on the womans procreation choices (i.e., trying to impose or convey a feeling of an incorrect nature of the actions being taken, to persuade a woman not to have children). Russian specialists turned out rather reluctant to reliably protect the patients confidentiality. For example, a considerable part of geneticists in Russia (about 60 %) strongly believed that bank-stored DNA samples should be readily available to the blood relatives of the patient without prior consent of the latter. Hence, the substantial part of Russian geneticists were inclined to both paternalistic decisions and directiveness. This was largely predetermined by insufficient legal regulation of the genetic services activity in Russia, the peculiarities of the system of organization of the genetic service, poor availability and accessibility of the facilities engaged in providing proper treatment of hereditary diseases, as well as medical and social rehabilitation.Based on the obtained findings, studying of ethical issues in medical genetics was included into the professional training programmes. P01.28 Genetic Counselling in Forensic Medicine

Genetic counselling is an essential area in healthcare. Educational programmes for non-physicians exist since 1969, in the USA. In Europe, the first started in 1992, in the UK (Manchester and Cardiff). In the last 5 years, new master courses began in Europe (Bergen, Marseille, Genoa, Barcelona, Groningen and Uppsala). In 2009, we initiated a two-year full-time master programme (120 ECTS). It is limited to 5-6 students (nurses, clinical psychologists), with some previous clinical experience. Its main objective is to train professional counsellors, to join multidisciplinary teams at medical genetic services and consultations. A structured curriculum was based on the ESHG core-competences for genetic counsellors. The course consists mainly of small-group tutorials and has a large practical component. Educational areas are (1) principles and practice of genetic counselling; (2) human and medical genetics; (3) communication skills, clinical psychology, mental health, and psychosocial genetics; (4) public health, community genetics, organization of services, health policy and health economy; (5) quantitative and qualitative methods and research methodologies; and (6) bioethics and medical ethics. Nuclear disciplines in these areas are required, as are clinical rotations (prenatal diagnosis, paediatrics, neurological disorders, cancer genetics, clinical psychology); a large range of optional disciplines and clinical rotations are also offered as part of the first curricular year. The second year is fully in training at a medical genetics unit, and includes a research seminar. We expect this to harmonize with other European programmes and help moving toward recognition of the profession of genetic counsellors in Portugal and in Europe. P01.30 Genetic counsellors and predictive medicine : the profession and her evolution in France

C. Cordier1,2, M. Voelckel3,2; 1 Department of Cytogenetic, Mulhouse, France, 2French Association of Genetic Counsellors, Marseille, France, 3Department of medical genetic, Marseille, France.

A. Kondo, I. Hasegawa, Y. Onuki, Y. Nishijima, M. Ikeda, H. Yokoyama, M. Mizoguchi, S. Izumi; Tokai University, Isehara, Japan.

Background: Genetic counselling is becoming more general medical service these days. In our country, this situation is same as other countries and Genetic Counsellor and Clinical Geneticist are increasing recently. However they are not enough for covering all patients still now. In fact there is no report of collaboration between Clinical Genetics and Forensic Medicine at present. Aim: To develop good relationship with Forensic Medicine and provide fair genetic support after sudden death of family member because of genetic disease. Methods: To elucidate the better situation of genetic counselling, we examine our previous two cases and estimate needs of genetic support from national and our Universitys sudden death statistics. Result: In our previous medical service system, it was difficult to make any contact with genetic staffs from the family members because forensic medicine is not a clinical department. However, at our University Hospital, we had 41(7.2%) sudden death cases related to genetic condition out of 569 sudden death in total (2005-2008). Most cases were male between 20s - 40s and had genetic heart condition (hypertrophic cardiomegaly, long QT syndrome, familial hypercholesterolemia, Burgada syndrome, etc), Marfan syndrome and Polycystic Kidney. Conclusion: For the bereaved family, we could provide more efficient medical service not only physically but also mentally. It will make the family members to see doctors for regular check up and avoid another future sudden death. It is very important that the collaboration between Genetics and Forensic medicine.

The professional master of Human Pathology, Genetic Counsellor and Predictive Medicine welcomed the first students of this speciality in France in the academic year 2004-2005. Today, 72 graduates have been trained to practise this new profession of health, which is governed by a law, in different genetics services throughout France and abroad. Using data from the French Association of Genetic Counsellors, we present this phase in the evolution of the dynamic and multidisciplinary profession of genetic counselling. This course has a vital role in the recruitment and training of these practitioners. The diversity of skills acquired through the course include, for examples, to establish relationship and to clarify patients concerns and expectations, to make appropriate and accurate genetic risk assessment, to work with a multidisciplinary health team, to integrate ethical and legal aspects, to use information systems, and to contribute to the research and education. Genetic counsellors have been integrated into multidisciplinary centers for prenatal diagnosis and are involved in consultations and the predictive diagnosis of assisted reproduction. Graduates are also working in different services like medical genetics and oncology, cardiology and neurology. We also assist in the efforts made by the french genetic counsellors to integrate national professionals in genetics into the FFGH (French Federation of Human Genetics) and the Group of Genetics and Cancer. French genetic counsellors are also participating with the ESHG and the European Network of Genetic Nurses and Counsellors to write the professional and educational standards for genetic counsellors in Europe.

Genetic counseling Genetics education, Genetic services, and Public policy

P01.31 Reporting on genetic disorders and syndromes in cyprus

V. Christophidou Anastasiadou1,2, E. S. Aristidou1, A. Kotti2, T. Delikurt1; 1 Clinical Genetics Service, Cyprus Institute of Neurology and Genetics, Nicosia, Cyprus, 2Clinical Genetics Service, Makarios Medical Center, Nicosia, Cyprus.

Cyprus is a small island, of 867,600 inhabitants, in the eastern Mediterranean. First human activity in 10,000 BC was followed by a Greek settlement (1400BC-1050 BC) and many invaders added to the genetic pool. This study reports on the diversity of diagnosis established for 3400 individuals in the years 1995-2009. A registry reports on medical and demographic data. The population, initially mainly Greek-Cypriots, few Turkish Cypriots, Maronites, and Armenians, gradually became of mixed descent or more multiethnic. Referrals included all common indications. 1007 patients were diagnosed. Monogenic disorders represented the largest category while chromosomal aberrations were less frequent. Neurofibromatosis type 1 and Trisomy 21 were the commonest accordingly. 96 patients were reported with multifactorial congenital anomalies and exposures accounted for 24 patients. 986 individuals were referred for genetic counseling for various indications (except Thalassemia which is managed by the National Thalassemia center). Presymptomatic genetic counseling and testing was offered to 290 individuals. Several rare and very rare syndromes were recognized. Discussion: Founders effect is evident in several communities and geographical areas. Congenital anomalies are under-represented probably due to the inconsistency in reporting and a high uptake of prenatal surveillance leading several affected pregnancies to termination. Genetic counselling initially was not appreciated but is now increasingly demanded. In conclusion, we are reporting on the evolution of clinical genetics services and the epidemiology of genetic disorders in Cyprus in the years 1995-2009. P01.32 the genetic testing offer in Europe: the need for crossborder testing

a half-day workshop for general practitioner (GP) trainers involved in supporting the genetic component of the national curriculum for general practice training. Instead of delivering the content as a series of didactic lectures, the workshops were designed so that content was provided using interactive case studies. This allowed GPs to draw on their own clinical experience, and highlighted where genetics was relevant in primary care practice. Sessions were evaluated using a multistaged approach, including evaluation forms immediately after the workshop (n=51), and follow-up questionnaires (n=17) and interviews (n=4) several months afterwards. Findings showed that the structure and delivery of the workshop served as a model for good teaching practice, with participants using similar interactive case studies in their own teaching sessions. In addition, participants reported how using the clinical scenarios in the workshops allowed reflection on their own practice, which has resulted in GPs changing their clinical management. All GPs trainers (n=17) who returned the follow-up questionnaire reporting changes in their clinical practice such as: changes to referral patterns; use of family history within consultations and improved communication with patients about genetic issues. The National Genetics Education and Development Centre is now offering this course to GP trainers throughout the UK. P01.34 Large normal (intermediate) [27-35 cAGs] and reduced penetrance [36-39 cAGs] alelles are not a rare event in Huntington disease (HD)
J. Sequeiros1,2, E. Ramos1, J. Cerqueira1, M. Costa1, A. Sousa1,2, J. PintoBasto1,2, I. Alonso1; 1 IBMC, Univ. Porto, Portugal, 2ICBAS, Univ. Porto, Portugal.

M. Jovanovic1, E. Dequeker2, L. Desmet2, M. Morris2, J. Cassiman2, S. Aym1; 1 Orphanet - INSERM SC11, Paris, France, 2EuroGentest, Leuven, Belgium.

Genetic tests are now offered internationally, through both public and private sector genetic testing services. Physicians prescribing these tests and biologists receiving the samples need to know which tests are available, where they are performed and whether identified laboratories meet quality standards. To fulfill this need, www.orpha.net was launched thirteen years ago to set up a database of clinical laboratories in the field of rare diseases. Data was collected in 1 country in 1997, 15 in 2003, 26 in 2006 and 38 in 2010, with resources from the EC DG Public Health. In collaboration with the EuroGentest Network of Excellence, information on quality management has been added to the Orphanet database over the past four years. Information on genetic testing in Orphanet can be searched by disease name or gene (symbol or name in English) as well as by laboratory or professional. The information provided on laboratories includes data on quality management. Currently, 956 laboratories offering tests for 1,559 genes are registered. The test offer differs greatly from one large country to another: Germany (1,141 genes), France (874 genes), Italy (625 genes), Spain (582 genes), UK (414 genes). Medium and small-sized countries test offer ranges from 1 to 233 genes. This situation explains the large cross-border flow of specimens, highlighting the need to provide access to services in other countries when necessary, especially for very rare diseases. According to available data, only testing for Cystic fibrosis is provided by every country. The distribution of this test offer will be presented. P01.33 Achieving change in clinical management: results from a genetics education intervention
M. Bishop1, S. Burke2, P. Farndon1; 1 NHS National Genetics Education and Development Centre, Birmingham, United Kingdom, 2Centre for Research in Medical and Dental Education, University of Birmingham, Birmingham, United Kingdom.

Four classes of alleles exist in HD, according to (CAG)n size; nevertheless, only two outcomes (carrier or non-carrier) are usually discussed in genetic counselling, as the possible results of presymptomatic testing (PST). Though large normal alleles may produce de novo expansions, little is known about their prevalence and impact in daily practice. We estimated the frequency of large normal (class 2) and reduced penetrance (class 3) alleles, and the frequency of genotypes carrying them, in (1) our diagnostic laboratory (1,214 samples, 350 families), (2) our genetic counselling clinic (146 testees), and (3) our general population (2,000 control chromosomes). Large normal alleles were 6% of population control alleles, 7% of consultands for PST, and 7% of all diagnostic samples (they represented 5% of all normal HD alleles at the laboratory). Reduced-penetrance alleles were found in only one control chromosome (0.1% individuals), but in 5% of PST, and >2% of all lab samples (they were 4.3% of all expansions found, and >9% in PST). Evidence thus showed that large normal alleles are relatively frequent in the general population, while reduced penetrance alleles are also not rare at the lab or the clinic. Together, these two classes were present in >10% of all consultands for PST. The four existing classes of alleles must be addressed in pre-test counselling. If not aware of all possible test outcomes, consultands may become very distressed when learning that their results fall within a grey-zone, rather than providing a definite prognosis for themselves and/or their children (<90% cases). P01.35 clinical coding of Rare and Genetic Diseases

M. Milicic Brandt, M. Cornell, A. Devereau; National Genetics Reference Laboratory, Manchester, United Kingdom.

While there is a continual emphasis on the need to educate non-genetic specialists in genetics, does education actually make any difference to the clinical management of patients? To answer this question, the UK National Genetics Education and Development Centre developed

NGRL is participating in a clinical coding project that includes mapping the Orphanet (www.orpha.net) rare disease catalogue to three established medical terminologies, including SNOMED CT, in order to improve the representation and traceability of rare diseases in health records and systems. Orphanet medical terminology contains data on approximately 6,000 rare (largely genetic) diseases, relating clinical signs and genes to diseases, including epidemiological data, as well as classification of rare diseases. The Orphanet group are revising both their classification of rare diseases and the ICD-10 classification of rare diseases as part of this project and the development of ICD11. In order to enforce the consistency of Orphanet data we developed a logical (OWL) model and used automated reasoning to detect errors. For example, we identified instances where epidemiological constraints were inconsistent between a disease and its sub-categories,

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implying either errors in classification or in the epidemiological data. Ensuring that the classification of diseases is correct is of great importance for producing valid mappings between different terminologies and in ensuring the utility of the terminologies. The mapping process included several steps. The first, lexical mapping, based on syntactic comparison of disease names, provided us with a set of exact and partial matches. The second step included the quality control of these mappings by clinical experts, and the last step used structural data, such as classifications of diseases, to provide additional partial mappings. Our final goal is to revise SNOMED CTs data on rare diseases by using mappings from Orphanet to this terminology. P01.36 From newborn screening to primary prevention of hemoglobinopathies

speech: he is not the unique information provider and his patient has his own opinion about genetic information. We therefore aim to discuss the potential impact of this evolution on patient/clinician relation and its incidence about clinical practice. P01.38 The study of polymorphic variants in the serotonin receptor gene HtR2A have professional athletes.

A. Timkova, O. Gumerova, V. Gorbunova; Bashcir state pedagogical univercity from name it M.Akmulla, Ufa, Russian Federation.

J. O. Kaufmann1, I. Krapels2, B. Van Brussel1, R. C. Zekveld-Vroon3, J. C. Oosterwijk4, F. van Erp5, P. J. G. Zwijnenburg6, P. C. Giordano1; 1 LUMC, Leiden, Netherlands, 2Maastricht University, Maastricht, Netherlands, 3 Erasmus University, Rotterdam, Netherlands, 4University Medical Center Groningen, Groningen, Netherlands, 5University Medical Center St Radboud, Nijmegen, Netherlands, 6VuMC, Amsterdam, Netherlands.

Newborn screening for sickle cell disease (SCD) started in the Netherlands upon approval of the national health council. This was after growing numbers of patients were born in the Netherlands, due to increased immigration from endemic regions, their endogamous partner choice, and their relative high number of consanguine marriages. The first year of screening revealed a hemoglobinopathy in 64 children (0.35) 41 of which had SCD and 806 (4.2 ) was a HbS carrier. The detection of carriers of sickle cell offers parents the chance to prevent a possibly affected child in the next pregnancy. After indentifying a carrier the parents ought to be offered diagnostics and when both are carriers the couple at risk should be referred to a genetic center. Therefore, we monitored the hemoglobinopathy intakes at the genetic centers since the start of the newborn screening. We asked all 8 clinical genetic centers in the Netherlands for their HbP counselings and gave them a short questionnaire per case asking: reason for referral, ethnic background, diagnosis and plans with regards to family planning. We got cooperation of 7 centers that provided us with their anonymized data. Preliminary results show incidental referrals after newborn screening. Reasons for not seeing these families in the genetic centers may vary and remains unknown. One of the reasons maybe the pediatricians treating the affected children counsel these families themselves, but in a worse case scenario parents get no counseling and may be confronted with another ill child, without this being an informed choice. P01.37 clinical geneticists tomorrow: Are quantity & diversity of information an opportunity or a trap?
S. Julia1,2, A. Soulier1, A. Cambon-Thomsen1; 1 UMR 558 Inserm, Universit Paul Sabatier, Toulouse, France, 2Service de Gntique Mdicale, Hpital Purpan, CHU de Toulouse, Toulouse, France.

Important indicators of the success of sporting activity is a resistance to psychological stress, the ability to receive and processing of information, largely determined by the work of the neurotransmitter systems. One of the base genes neuromediator serotoninergic system is the serotonin receptor gene HTR2A localized on 13.q -14 - q 21 chromosome. The analysis of frequency distribution of alleles and genotypes of the serotonin receptor in the group of sportsmans. The analysis of the frequency distribution of alleles and genotypes of the serotonin receptor in the group of sportsmen. The analiz polymorphism A1438 in the gene HTR2A conducted in 250 unrelated individuals by PCR with subsequent processing amplifikons appropriate restriction endonuclease. Of these, 150 people who are not involved in sports, and 100 human professional athletes. Found a significant increase in the frequency of allele G (P = 0.01) and genotype GG (P = 0.005) in the group of athletes on the background of reducing the frequency of allele A (P = 0.002) and AA genotype (P = 0.005) in the comparison group.Allele G polymorphic locus HTR2A in gene A1438G have may a positive potential for sports activities.This work was partially funded grant from the Ministry of Education of Russia Thematic Plan for 2008-2010. P01.39 Psychological well being and quality of life in presymptomatic Huntingtons Disease gene carriers.

S. Serpentini1, L. Dal Sasso1, P. Castellan1, E. Pasquin1, M. Balestrieri2, G. Damante1, I. R. Lonigro1,3; 1 Istituto di Genetica - A.O.U., Udine, Italy, 2Clinica Psichiatrica - A.O.U., Udine, Italy, 3Dipartimento di Scienze e Tecnologie Biomediche - Universit, Udine, Italy.

Clinical geneticists have to interpret genetic data coming from the laboratory and to position themselves in the information society. Their interpretative task tightly depends on 1. the nature of data provided by genetic testing technologies and 2. the knowledge of the patient. Patients can indeed access information and their own understanding of genetics is a relevant parameter in the patient/clinician communication. Through the development of new technologies - both in the laboratory and in the society - clinical geneticists will therefore have to face new kinds and new sources of information. Rapid developments of high throughput technology (HTT) let expect that new genetic testing technologies will be implemented soon in clinical laboratories. Sequencing genes generates quantitatively abundance of data and qualitatively complex and heterogeneous information with different levels of clinical relevance. As data interpretation becomes tougher and unexpected health related information are more common, emerging technologies will obviously challenge medical practices. At the same time as clinicians deal with growing uncertainty, new technologies of communication provide diverse sources of information to patients. Networks strengthen patients communities, websites make genetic testing available directly to consumers and information flows literally on line. Diversity of information challenges the clinicians

Purpose: The aim of the study is to describe some preliminary data about psychological well being and quality of life in presymthomatic Huntingtons Disease (HD) gene carriers after 6-12 months from DNA testing. The study is part of a larger, long-term research and has only a descriptive purpose because of the small number of the sample. Methods: From May to December 2009, at the Institute of Genetics of Hospital/University S. Maria della Misericordia of Udine (Italy), we evaluated 10 presymthomatic subjects resulted carriers of HD gene. The instruments used are: 1) a semi-structured interview; 2) the Psychological General Well-being Index (PWBGI); 3) the Short Form 36 Health Survey Questionnaire (SF-36). Results: Until now we assessed 10 patients (M:4, F: 6; mean age: 50,20, range age:28-75). The means and standard deviations of the PWBGI scales are: Anxiety 15,60 (4,90), Depression 10,80 (2,57), Well-being 10,40 (3,56), Self control 9,70 (3,53), Global health 9,60 (4,55), Vitality 10,20 (4,18). The means and standard deviations of the SF-36 scales are: Physical functioning 70 (30,83), Role-Physical 47,5 (50,62), Bodily pain 78,6 (30,13), General health 53,22 (31,66), Vitality 51,11 (25,34), Social functioning 72,50 (28,75), Role-emotional 60,00 (51,64), Mental health 62,22 (18,98). Our mean scores found to be lower than normative scores. Conclusions: Our preliminary descriptive results indicate a critical condition in the psychological well being and the quality of life of the presympotamic HD gene carriers and suggest the importance of a specific psychological intervention. Therefore, we consider necessary to continue this study with a larger sample. P01.40 An analysis of the impact of illness representation in predicting distress in breast cancer patients; implications for genetic counselling
R. Moldovan; Babes-Bolyai University, Cluj Napoca, Romania.

The role played by psychological, social and cultural factors in health and illness related behavioral and emotional responses is no longer breaking news. Individual differences such as various cognitions have been shown to contribute in variability in distress when confronted with

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stressful events or certain illnesses. Still, there is considerable lack of empirical data concerning the impact that illness representation has in predicting distress for patients diagnosed with breast cancer. The purpose of this mixed methods study was to investigate the interrelations between illness representation and distress levels in breast cancer patients. The illness representation of 30 breast cancer patients was evaluated with semi-structured interviews; distressed was assessed using the Beck Depression Inventory, State Trait Anxiety Inventory and the Profile of Mood States. Results revealed no significant differences between levels of distress for different representations of illness (p>0.05). Implications for genetic counselling are discussed. P01.41 Genetic causes of intellectual disability
G. Cozaru, C. Papari; Andrei aguna University, Constanta, Romania.

community acceptance of depression-risk genotyping, even though a predisposition to depression may only manifest upon exposure to stressful life events. Our results suggest there will be a strong demand for predictive genetic testing for complex multifactorial disorders. P01.43 Genetic counselling and testing in inherited monogenic cardiac disorders : interviews with patients and their families

S. Fokstuen1, D. Kumar2, J. L. Blouin1, A. Clarke2; 1 Genetic Medicine, Geneva, Switzerland, 2Institute of Medical Genetics, Cardiff, United Kingdom.

This paper reviews several genetic syndromes that are associated with intellectual disability (ID). ID is a condition characterized by significantly sub-average intellectual functioning (often expressed as I.Q. < 70 to 75) combined with limitations of > 2 of the following: communication, self-direction, social skills, self-care, use of community resources, and maintenance of personal safety. Between 1 % and 3 % of a population have an intellectual disability. Management consists of education, family counseling, and social support. We want to present 5 cases: Cri du chat syndrome, Down syndrome, Turner syndrome, Coffin-Lowry syndrome and Sotos syndrome. All this cases have in common: delayed intellectual development, immature behavior, limited self-care skills, but genetic causes are different. All children with an intellectual disability (mental retardation) or global developmental delay should have a comprehensive evaluation to establish the etiology of the disability. Family support and counseling are crucial. As soon as ID is confirmed or strongly suspected, the parents should be informed and given ample time to discuss causes, effects, prognosis, genetic recurrence risk, education and training of the child, and the importance of balancing known prognostic risks against negative self-fulfilling prophecies in which diminished expectations result in poor functional outcomes later in life. A diagnosis also avoids unnecessary testing and can lead to opportunities for improved health and functional outcomes. Finally, early comprehensive prenatal care and preventive measures prior to and during pregnancy increase a womans chances of preventing intellectual disability. P01.42** Public interest in depression-risk genotyping in a large national sample.

Experience with genetic counselling and testing for monogenic cardiac disorders is limited. What has been learned from dealing with other conditions like Huntingtons disease or familial cancer disorders may be applicable in these families too but the context of inherited cardiac conditions present very particular problems, especially sudden cardiac death in young adults, restrictions placed on exercise and uncertainty regarding the prognosis. In addition to the advantages and disadvantages of diagnostic testing from a clinical perspective, there are issues of privacy or disclosure and coping within the family with the risk of transmission and the possible psychological consequences. One issue that requires particular attention is how families experience and cope with being told that several of them may die at any moment. Problems with the transmission of such information take different forms depending upon whether the information is to be passed vertically to a child or horizontally to a sibling or other relative and also depending upon the availability of preventive and/or therapeutic strategies. These issues were discussed in a small number of interviews with patients and/or their families, who have experienced cardiac death due to an inherited cardiomyopathy in a young adult first degree relative. We present their comments about genetic counselling and genetic testing. It clearly emerges that genetic counselling is a key component in the overall management of families affected with inherited monogenic cardiac disorders and that more research in the personal experience of these families is necessary to improve medical management and support. P01.44 Genetic counselling and testing in cardiomyopathies

N. Marziliano, P. Merlini, M. Frigerio, G. Masciocco, S. Veronese, F. Orsini, A. Colosimo, C. Lauricella, M. Gambacorta, F. Mauri; Azienda Ospedaliera Ospedale C Granda Niguarda, MILANO, Italy.

A. Wilde1,2, B. Meiser3, P. B. Mitchell1,2,4, D. Hadzi-Pavlovic1,2, P. R. Schofield5,4,6; 1 School of Psychiatry, University of New South Wales, Sydney, NSW, Australia, 2 Black Dog Institute, Sydney, Australia, 3Prince of Wales Clinical School, University of New South Wales, Sydney, NSW, Australia, 4Brain Sciences UNSW, Sydney, Australia, 5Prince of Wales Medical Research Institute, Sydney, NSW, Australia, 6School of Medical Sciences, University of New South Wales, Sydney, Australia.

Introduction: Despite international concern about unregulated predictive genetic testing, there is surprisingly little data on both the determinants of community interest in such testing and its psychosocial impacts. Methods: A large population-based public survey with community dwelling adults (N=1046) ascertained through random digit dialling. Attitudes were assessed via structured interviews. Results: The study found strong interest in predictive genetic testing for a reported susceptibility to major depression. Once perceived benefits and disadvantages of such testing had been considered, there was significantly greater interest in seeking such a test through a health care provider (62%) compared to direct-to-consumer (40%) (p<0.001). Personal history of mental illness (OR= 2.53, p<0.001); self-estimation of being at higher than average risk for depression (OR = 1.84, p<0.001); belief that evidence of genetic component for a mental illness would increase rather than decrease stigma (OR=1.51, p<0.001); sexual abuse as a casual attribution for depression (OR = 1.24, p <0.001) and endorsement of perceived benefits of genetic testing (OR=3.18, p<0.001) significantly predicted interest in having such a test. Conclusions: Despite finding attitudes that genetic links to mental illness would increase rather than decrease stigma, we found strong

The recent and major breakthrough in the molecular genetics of Cardiomyopathies has created a new understanding of these diseases, and also new expectations regarding the applications in clinical practice. A new task for Cardiologists and geneticists in the cardiovascular settings is therefore to integrate this new knowledge in order to improve the management of the patients and their families. Beyond the traditional goal of genetic counselling, defined as a communication process which deals with the human and psychological problems associated with the occurrence or the risk of occurrence of a genetic disorder in a family, the main goals are (i) to inform the patients and family about the genetic aspects of the disease, including the risk of transmitting the disease within the family; (ii) organise appropriate cardiac screening and follow-up of the relatives; (iii) discuss genetic testing, which may improve the medical management in various situations. By showing a clinical and diagnostic work-up of a family with a compound heterozigosity in the MYBPC3 gene, these objectives are achieved taken into account the specific issues related to a genetic disease, such as the psychological, social, professional, ethical and legal implications. Genetic counselling and genetic testing should therefore be performed by trained health care providers and usually through a close collaboration between different and complimentary specialties. P01.45 the neonatal screening programme in Leningrad province: 2009

I. A. Ivanov1, M. O. Mkheidze2,1, E. S. Davidova1, I. F. Mamontova1, I. M. Andreeva1, S. E. Khalchitsky1; 1 District Children Hospital, St.Petersburg, Russian Federation, 2Medical Academy for Postgraduate Studies, St.Petersburg, Russian Federation.

Genetic neonatal screening is an important public health and clinical activity. In Leningrad province the neonatal screening programme was expanded from 2 to 5 disorders for which screening met the Wilson and Jungner criteria, especially regarding treatability. In 2009 Labora-

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tory of molecular genetics and cytogenetics (LMGC) as a unit of the District Children Hospital realized neonatal screening for phenylketonuria (PKU), galactosemia (Gal), congenital hypothyroidism (CH), congenital adrenal hyperplasia (CAH), cystic fibrosis (CF). Guthrie blanks spotted with newborns blood (12800 infants) were analyzed with automatic fluorescent methods and fluoroimmunoassay. The PKU-neoscreen reagents (ZAO Immunoscreen Moscow, Russia) were used in PKU screening. Screening for Gal was realized with Neonatal Total Galactose kit (PerkinElmer.Inc). The Delfia Neonatal hTSH assay was used in CH screening. A risk group for CAH was formed according to the data of the Delfia Neonatal 17--OH-progesterone kit. CF screening was based on the Delfia Neonatal IRT assay. Two infants had constantly high levels of Phe and were found to be PKU infants. 30 babies were found to be a risk group of CH. 17 infants formed a risk group of CAH. Nobody was shown to be included in risk lists of Gal and CF. Clinical geneticists of LMGC confirm hereditary disorders, carry out long-term outpatient care, dietary management and genetic counselling P01.46 Newborn hearing screening

the groups of the subjects (100%) agreed that the destruction of an embryo prior to implantation less wrong than destruction of the fetus inside the uterus in comparison to only 30% of the PGD group. Views were more mixed for the other (IVF) procedure. Conclusion The outcomes of this study demonstrate that PGD might be considered as a valid alternative to prenatal diagnosis. However, couples referred for PGD must be selected and counselled appropriately, considering the complexity of the treatment and the relatively low take-home baby rate. P01.48 The structure of the public attitude toward the genome research by the latent class analysis in Japan

Z. Yamagata1, T. Maeda2, K. Muto3, A. Nagai1, A. Tamakoshi4, I. Ishiyama5; 1 Department of Health Sciences, School of Medicine, University of Yamanashi, Chuo, Yamanashi, Japan, 2The Institute of Statistical Mathematics, Tokyo, Japan, 3The Institute of Medical Science, The University of Tokyo, tokyo, Japan, 4Aichi Medical University, Aichi, Japan, 5Teikyo-Gakuen Junior College, Yamanashi, Japan.

E. Severin1, A. Toma2, C. Albu1, D. Albu1, C. Sarafoleanu1, C. Dragomir3, A. Stan3, D. T. Stefanescu3, L. Savu3; 1 Carol Davila Univ Med Pharm, Bucharest, Romania, 2Panait Sirbu Hospital, Bucharest, Romania, 3Genetic Lab, Bucharest, Romania.

Background: newborn hearing screening has been instituted in many countries since the identification of congenital hearing loss in the neonate period, followed by intervention within the first few months, is critical for limited communication, low school achievements and life-long dependency. In the absence of newborn screening, the hearing loss might not be noticed by parents until the child begins to have difficulties at speaking and learning at a later age. Approximately 90% of children with deafness are born to hearing parents. Most cases with non-syndromic deafness inherited the disorder in an autosomal recessive manner. Aims of the study: identification of congenital hearing loss in children before three months of age; detection of risk factors known to be associated with congenital hearing loss. Methods and Subjects: Otoacoustic emissions (OAEs) and auditory brainstem response (ABR) were performed for 4303 newborns. Physical examinations and family history were used to get information about hereditary / syndromic hearing loss. Results: 23 newborns did not pass the OAEs and ABR. All of them were referred to audiologic evaluation to confirm if the hearing loss is present. Physical examination revealed no other findings associated with a syndrome. Four newborns with hearing parents were found to have sensorineural hearing loss. They were referred to genetic counseling and testing for the etiologic diagnosis. Conclusion: screening newborns for hearing loss is highly accurate and leads to earlier identification and treatment of infants with hearing loss. All newborns identified with congenital hearing loss require a comprehensive genetic evaluation. P01.47 Pre-implantation Genetic Diagnosis in saudi Arabia: Parents Experience and Attitudes

The purpose of this study was to clarify the structure of the public attitude toward the genome research by the latent class analysis. The nationalwide surveys about the attitude toward the genome research were conducted in 2005 and 2009 in Japan. The participants were comprised of 4,000 people (age, 20-69) in 2005 and in 2009, selected from the Japanese general population by using the two-step stratified random sampling method. Five clusters were assumed as an explanation model of six variables related to the knowledge of genome and attitudes toward genomic research promotion about three themes; basic genome research, genome research related to agriculture and medicine at the survey in 2005. They were able to be named Group of aggressive promotion (40.8%), Group of passive support (20.2%), Group not making judgment (18.5), Group making prudent judgment (16.5%), and Group not interested in genome. The results in 2009 were almost the same as that in 2005. It is possible to forecast to which cluster to belong according to respondents attribute, and we can forecast the reaction to other questions by using a cluster oppositely. For examples, Group of aggressive promotion is the layer of a high academic background, and is positive to donate their blood for the genome research, Group making prudent judgment is high academic background persons as same as Group of aggressive promotion, and is interesting in the science and technology, but is negative to the blood donation for the research. P01.49 cross-disease support group

R. Rupps1,2, C. J. Condin3, I. Jordan2, C. du Souich1,2, C. F. Boerkoel1,2, W. H. McKellin3,2; 1 Department of Medical Genetics, University of British Columbia, Vancouver, BC, Canada, 2Rare Disease Foundation, Vancouver, BC, Canada, 3Department of Anthropology, University of British Columbia, Vancouver, BC, Canada.

A. Al Suliaman; Department of Genetics, Research Centre, King Faisal Hospital and Research Centre, Riyadh, Saudi Arabia.

Background preimplantation genetic diagnosis (PGD) has been proposed as a valuable alternative to prenatal diagnosis (PND) to select genetically normal human embryos and transfer them to the uterus of a woman. This study evaluates a range of social and moral concerns of Saudi couples with and without experience of the IVF procedure might have about the various procedures available. Methods A total of 184 subjects attending the King Faisal Specialist Hospital and Research Centre in Riyadh were interviewed using a semi-structured questionnaire. 87 of the subjects have complete at least one cycle of the in vitro fertilization (IVF) procedure. Results Forty-nine (100%) of the oncology group and forty-three (90%) of the ENT group were personally accept PGD technology. All the oncology and ENT group who would personally consider PGD were willing for others to be offered the procedure. Specific concerns about PGD related to the IVF procedure, waiting for the pregnancy results and egg collection were the most commonly mentioned concerns. All

Background: Rare diseases are individually uncommon (affecting fewer than 1 in 2000) but collectively affect approximately 1 in 10. Parents of children with very rare and undiagnosed diseases often feel hopeless and alone. Hypothesis: Support addressing the needs and isolation of individuals and families with very rare and undiagnosed diseases is possible within a diverse community. Results: To test this hypothesis, we profiled a series of individuals and families with very rare and undiagnosed diseases. Shared features that we identified included sense of isolation, need for story-sharing and desire for more information. Distinguishing features that we identified included parents experiencing greater anxiety with new compared to more established diagnoses and having an undiagnosed compared to a known condition. The parents identified common themes of basic medical, educational, social and advocacy needs. Based on these commonalities, we formed a trial support group composed of 19 families with 18 diagnoses. Over the course of 2 years, our observations are that it is possible to provide cross-disease support on issues relevant to a variety of disorders. Conclusion: Cross-disease support groups are very effective for addressing the shared medical, educational, social, advocacy, and research needs of the community with very rare and undiagnosed diseases. Our model allows universal establishment of such local support groups

Genetic counseling Genetics education, Genetic services, and Public policy

P01.50 WHO International Classification of Diseases (ICD) Revision Process: state of Art for genetic diseases
S. Aym, B. Bellet, A. Rath; Orphanet - INSERM SC11, Paris, France.

WHO has established various Topic Advisory Groups to serve as planning and coordinating advisory bodies in the update and revision process for specific areas of the ICD. A TAG for rare diseases (including all genetic diseases) was established in April 2007 as rare diseases should now be traceable in mortality and morbidity information systems. The production of the basic information needed to establish an Alpha draft of the classification of rare diseases has been assigned to Orphanet. The Orphanet database includes over 6,000 distinct phenotypes which are classified according to published classifications. These classification systems are mainly based on scientific grounds (aetiology and mechanism). Orphanet has developed a complementary, strictly clinical in-house classification to meet clinicians needs. Available on Orphanets website, this classification serves to elaborate an ICD revision proposal. The revised chapters currently circulating among experts for review are Haematology, Endocrinology, Nutrition, Metabolism and Immunology, Neurology and Malformation. Revised chapters follow a primarily clinical approach, only secondarily an aetiological one up to the gene level. When several names are possible for a disease, descriptive names formed in accordance with a clinical approach are preferred. Every entity is assigned a unique identifying number. Rare diseases affecting several body systems are included in every relevant chapter, as ICD11 will be polyaxial: a main code is also selected for linearisation purposes, according to the severest involvement and/or the specialist most likely to manage the disease. In some cases, the choice is open for debate. Input from the Rare Disease Community is expected. P01.51 An overview of rare diseases research in Europe based on data from the Orphanet database.
N. Martin, N. Doulet, V. Hivert, S. Aym; Orphanet - INSERM SC11, Paris, France.

tion allows for best management of at-risk individuals and prevents unnecessary examination under anaesthetic (EUA), which can be traumatic for the child and family. This review identified families who had attended West of Scotland genetics over the past twenty years with unilateral or bilateral retinoblastoma. We identified 12 families who had stored DNA samples but not had complete analysis. The samples were sent for analysis of RB1 by bi-directional sequencing and MLPA. Germline mutations were identified in six bilateral cases, including one mosaic, and testing is now available to relatives. One bilateral case requires additional analysis by RT-PCR. No mutation was identified in the five individuals with unilateral retinoblastoma and no other known family history. Unnecessary EUA may now be avoided in their relatives and their families reassured. These families highlight the challenges of rare conditions, where results may take many years and families may need support to adapt to new information many years after diagnosis. P01.53 conceptual frameworks determine experts views on clinical use of schizophrenia genetics results

T. Vrijenhoek1, C. van El2, H. G. Brunner1, A. Geurts van Kessel1, M. C. Cornel2, J. A. Veltman1, A. Nelis3; 1 Department of Human Genetics, Radboud University Nijmegen Medical Centre, Nijmegen Centre for Molecular Life Sciences, Nijmegen, Netherlands, 2 Department of Clinical Genetics and EMGO Institute for Health and Care Research, VU University Medical Center, Amsterdam, Netherlands, 3Centre for Society and Genomics (CSG), Institute for Science, Innovation and Society, Radboud University Nijmegen, Nijmegen, Netherlands.

Orphanet compiles 6500 research projects. This information been analysed to identify areas needing collaborative research projects and to target future calls for proposals. The analysis of the distribution of number of diseases by number of treatments in development shows that most RD have no more than 3 products in development, whereas 53 RD have over three such products. Similar analysis results were obtained for clinical trials, marketed drugs, patient registries and preclinical/epidemiological/basic research. Some of the diseases overrepresented upstream in the process of R&D (with a treatment on the market or drugs in development) are well represented regarding ongoing research (Cystic fibrosis, pulmonary arterial hypertension, some rare cancers). Diseases with a higher prevalence are anticipated to have more treatments in development: this assumption is not backed up by our data analysis. The most developed areas where the ones for which large European networks and consortium have been established as it is the only way to achieve the critical mass necessary in terms of resources and expertise. However the sustainability of the structures which have been created during the development of these collaborations, such as patient registries, biobanks, and technological platforms, is very difficult to ensure to date. The EC could be involved in the financing of the coordination and maintenance of these structures through a specific call for proposals. This work was supported by the European Unions Seventh Framework Programme (HEALTHF2-2008-201230). P01.52 Review of Retinoblastoma families and RB1 mutational analysis in the West of scotland genetics service
S. Gibson, V. Murday; Ferguson Smith Centre for Clinical Genetics, Glasgow, United Kingdom.

Recent developments in microarray technology have enabled largescale genetic studies on schizophrenia, and have led to the discovery of many risk alleles. This development has been accompanied by promising expectations regarding clinical applications. However, most of the known genetic variants meet validity nor utility requirements for application in clinical practice. Ongoing discussions are not only about when or under what conditions, but also if results from large-scale schizophrenia genetics studies are relevant for clinical use. Based on a literature review and in-depth interviews with experts, we explored the arguments used in these discussions. We have identified four different lines of arguing, which state that high-throughput schizophrenia studies will: (1) eventually reveal clinically relevant data; (2) not reveal clinically relevant data directly; (3) only reveal clinically relevant data when combined with non-genetic (environmental) factors; or (4) eventually be translated into a clinical setting, irrespective of clinical relevance. We show that the experts perception of the relevance of large scale genetic data predominantly depends on their conceptual framework of the relationship between genes and schizophrenia. This is exemplified by discussions around the development of direct-to-consumer (DTC) genetic tests. Furthermore, the experts conceptual framework determines their perception of schizophrenia itself, which is partly reflected in recent discussions on DSM-V. The challenges resulting from large scale genetic research and the demand from consumers and patients for genetic testing call for a solid infrastructural and translational strategy for schizophrenia genetic research. P01.54 screening tests for Genetic counselling in tURKEY
Y. Erdem1, F. Teksen2; 1 Ankara University Faculty of Health Sciences, Ankara, Turkey, 2Ankara University Faculty of Medicine, Ankara, Turkey.

Rare inherited diseases can cause challenges for genetic services, both clinical and molecular. Retinoblastoma is a rare childhood cancer of the eye. Bilateral retinoblastoma is caused by de novo or inherited germline RB1 mutations. Most unilateral cases are due to somatic mutations. As the germline and somatic forms cannot be distinguished clinically, molecular diagnosis is essential for affected families. It is important that testing is conducted by an experienced laboratory; that results are given to families in a timely manner and at-risk relatives are identified in order to target screening. Identification of a genetic muta-

Turkey is a country where consanguineous marriages are quite common. Approximately of one-fourth of marriages in Turkey is consanguineous and as a result, the incidence of autosomal recessive diseases are very high. In order to avoid the recurrence of these diseases, Ministry of Health has started screening programmes. For instance, in 1995, a premarital screening programme aiming to reduce the incidence of thalassemia major at 33 city center; in 2002, Phenylketonuria (PKU) Screening Programme; in 2003, national newborn hearing screening program (NNHSP); in 2006, congenital hypothyroidism screening program; in 2008, a metabolic disorder, Biotinidase screening programmes were established. Beta-thalassemia prevalence in the country is 2.1%. There are approximately 1.300.000 carriers and 4000 patients in Turkey. The frequency of PKU is 1 in 4500 live births and 1 in 10,000 live births in the United States comparatively. The newborn hearing, PKU, hypothyroidism, and Biotinidase screening programmes were applied nearly to 95.1% of all

Genetic counseling Genetics education, Genetic services, and Public policy

newborns in last five years according to Ministry of Health report. In conclusion, it was observed that, premarital screening is a very useful tool for detecting carrier couples and an effective way of controlling the recurrence of some of the autosomal recessive genetic diseases. Also, there is the necessity to newborn screening programmes. In addition to screening programmes, the establishment of Genetic Counseling Services is also very important and the employment of high quality educated genetic counselors in these centers is of great value as they have critical roles in the prevention of inherited diseases. P01.55 Prevalent mutant alleles in connexin 26 ( GJB2 ) to romanian children with autosomal recessive nonsyndromic deafness

P01.57 Providing access to genetic variation: The SNP & SEQ technology Platform at Uppsala University

T. Axelsson, L. Bckstrm, E. Englund, C. Enstrm, O. Karlberg, K. Larsson, U. Liljedahl, M. Lindersson, P. Lundmark, D. Magnusdottir, C. Pntinen, I. . Thorsteinsson, A. Wiman, T. st, A. Syvnen; Uppsala University, Uppsala, Sweden.

C. Strugaru1, L. Bohaltea1, R. Calarasu2, A. Branzan3; 1 Department of Genetics Carol Davila University of Medicine and Pharmacy, Bucharest,, Romania, 2Department of Oto-rhino-laringology - Prof. Dr. Dorin Hociota Phonoaudiology and Laringology Hospital, Bucharest,, Romania, 3 Department of Pediatrics Dr. Victor Babes Diagnostic & Treatment Center, Bucharest,, Romania.

Inherited hearing impairment affects about 1 in 1000 newborns. Up to 50 percent of all patients with autosomal recessive nonsyndromic deafness in different populations have mutations in the gene encoding the gap-junction protein connexin 26 ( GJB2 ) at locus DFNB1 on chromosome 13q12. We performed mutation screening of 50 probands for GJB2 in nonsyndromic hearing loss families, including those with cases of sporadic deafness, which were compatible with recessive inheritance. 35delG and 167delT are two GJB2 alleles that cause nonsyndromic recessive deafness, and carrier rates for these mutant alleles may be as high as 4% in some ethnic populations (Estivill et al. 1998; Morell et al.1998). However, a large fraction (20 to 63 percent) of patients with GJB2 mutations have only one mutant allele; the accompanying mutation has not been identified. In our study, these results highlight the usefulness of Cx26 mutation screening for genetic counseling and suggest importance of entire sequencing of the gene responsible for DNFB1. P01.56 A practice framework for promoting appropriate reporting and use of molecular genetic test results: combining education, test result reporting, and information resources

The SNP & SEQ Technology Platform (www.genotyping.se) performs SNP genotyping and next generation DNA sequencing (NGS) as a service to academic groups in Sweden and the other Nordic countries and as partner in international collaborative projects. The platform offers genotyping services to research projects from single SNP-markers to genome-wide panels of 1 million SNPs in hundreds or thousands of samples, both for association studies and analysis of copy number alterations and DNA-methylation. The genotyping services include all steps from target selection and assay design, to production scale genotyping and quality assessment of the data. Small panels of 112 SNPs are analyzed by fluorescent single base primer extension, medium multiplex genotyping is performed by the Golden Gate assay, and highly multiplexed and genome-wide genotyping by the Infinium II assay (Illumina). Sequencing services are provided for all NGS-applications using two Genome AnalyzerIIx instruments (Illumina). We have developed a lab-data system for handling and storing information on samples, markers and genotypes, and for quality control and delivery of the genotype data. The genotyping process is accredited according to the European ISO/IEC 17025:2005 quality standard. For storing and analyzing NGS data using computer cluster hardware we collaborate with UPPMAX (Uppsala Multidisciplinary Center for Advanced Computational Science). Most of the completed genotyping and sequencing projects have studied human complex diseases but several projects in other species in evolutionary studies and comparative genomics have also been conducted. For a complete list of publications to which the SNP & SEQ Technology Platform has contributed, see www.genotyping.se. P01.58 Ethical Aspects of Prenatal Testing for Neurodegenerative and Nervous-muscle Hereditary Diseases in sakha (Yakutia
S. K. Kononova1, O. G. Sidorova1, V. L. Izhevskaya2; 1 Yakut Research Center of Complex Medical Problems, Siberian Branch of RAMS, Yakutsk, Russian Federation, 2Research Centre for Medical Genetics, Russian Academy of Medical Sciences, Moscow, Russian Federation.

I. M. Lubin1, M. Scheuner2,3, J. V. Henderson4, W. A. Faucett1,5, L. Hilborne2; 1 Centers for Disease Control and Prevention, Atlanta, GA, United States, 2 RAND Corporation, Santa Monica, CA, United States, 3VA Greater Los Angeles Healthcare System, Los Angeles, Los Angeles, CA, United States, 4 Dartmouth Medical School, Dartmouth, NH, United States, 5Emory University School of Medicine, Atlanta, GA, United States.

Clinical molecular genetic testing for heritable conditions is a growing segment of laboratory medicine. However, studies suggest that clinicians are challenged in staying up to date with knowledge about the genetic tests used in their medical practices and this increases the risk that patient care will be compromised. To address this concern, we propose a practice model to promote appropriate clinical decision making that combines education, effective laboratory reporting of test results, and access to credible and useful information relevant to the test ordered and result reported. This model is based upon several studies that are now complete and will be summarized. Some of the issues that the model addresses include 1) variations in the use of terminology and nomenclature, 2) understanding the relevance of the indication(s) for testing, 3) effectively communicating the limitations of the test method and result interpretation, 4) integrating knowledge of family history, race ethnicity, and other factors that are necessary for interpreting the test result, and 5) using the test result report as a tool for clinical decision making. An interactive, multimedia education program targeting clinicians has been developed to support this practice model. We facilitated collaboration between clinicians and laboratory professionals who evaluated test result reporting processes and developed a clinician-friendly laboratory test result report that has been adapted to a number of clinical scenarios. Combining these tools with access to readily available information resources should prove useful to practicing clinicians by enhancing professional competency and promoting good clinical decision making.

Genetic counseling of total 38 pregnant women at risk for spinocerebellar ataxia type 1 (SCAI, n=26) and muscular dystrophy (MD, n=12) have been carried out. All individuals were sakha nationality; women at risk for SCAI significantly more often had higher education than women at risk for MD (76.9% and 33.3%, p<0.05). Also, pregnant women at risk for SCAI were more likely than individuals with MD to perceive that prenatal diagnostics would influence their life. In SCAI group 14(53.8%) women requested prenatal testing then in MD group -10 (83,3%). Pregnant women at risk for SCAI more often didnt want prenatal testing than in MD group (46.2% and 16.2% respectively, p<0.05). The relation of women at risk to prenatal diagnostics can be caused a number of the reasons. The first reason is poor information about prenatal diagnostics, confirmed by questionnaires data of the sakha rural population. The second one is financial reason, so as not every rural family has a possibility to get to Yakutsk for a genetic counseling and DNA testing. Another possible reason is the ethnical features of sakha character: in interpretation of such a philosophical category as a destiny, that is patience in confronting of destinys strokes such as hereditary disease transmitting in the generation. P01.59 Teaching genetic counseling to first year medical students
R. Dragotoiu; Medical and Pharmacy University Carol Davila, Bucuresti, Romania.

Should genetic counseling be taught or is it encompassed in the overall ethical concepts first year medical students bear among their knowledge? Philosophers have assumed that morality is an inborn human trait and science has nowadays established the cortical regions which are activated while making explicit right or wrong judgments as well as while facing situations containing implicit moral issues. Whether a result of specific inborn brain development or interneuronal connections, a question raises: Can genetic counseling issues prove that

Genetic counseling Genetics education, Genetic services, and Public policy

their ethical aspects show a tendency of being either inherited or already gained through learning until becoming a student? After having read an article about the discovery of the CF gene, students had to answer 3 questions motivating their opinion. They were asked if considering right or not, that persons with genetic disorders took part in the researches concerning their disease, acknowledging that genetic disorders are rare and often severe; then, if in case of monogenic diseases with known gene anomalies they believed (thought) a screening in the teaching units to detect carriers of autosomal or X-linked genes could now cause in Romania unhappiness, discomfort, panic, because those persons would feel labeled as ill in front of friends and society, in general; lastly, if they considered useful or not the premarital carrier state detection. No discussions had taken place about genetic counseling during the workshops (and lectures), the results reflecting only the students personal views. The pitfalls of the study and the students answers are discussed for asserting the conclusion. P01.60 co-presence of deletional mutations on alpha globin genes in the beta thalassemia carriers in tehran

tized (M=2.39) if carrier status would be identified. Awareness and attitude towards carrier testing explained future reproductive choice (R2=.21; p < .001). Conclusion: All responding mothers showed high interest in receiving information on both thalassemia and carrier testing. The less educated and/ or the more deprived they were, the keener they were on receiving this information. Awareness of thalassemia was low. Even mothers with affected children seemed unaware of the inheritance pattern of the disease, showing the need for genetic counseling in Indonesia. It is therefore advised not only to raise awareness on thalassemia, but to better educate healthcare professionals as well. P01.62** Usefulness of Factor V Leiden mutation testing in clinical practice

E. . Blinkenberg1, A. Kristoffersen2, S. Sandberg2, V. M. Steen1,3, G. Houge1,3; 1 Center for Medical Genetics and Molecular Medicine, Haukeland University Hospital, Norway, NO-5021 Bergen, Norway, 2Laboratory of Clinical Biochemistry, Haukeland University Hospital, Norway, NO-5021 Bergen, Norway, 3Department of Clinical Medicine, University of Bergen, Norway.

B. Zarbakhsh, E. Farshadi, F. Eghbalpour, m. Mohammadi, A. Amirian, M. Jafari, S. Jamali, S. Zeinali; Pasteur Institute of Iran, Tehran, Islamic Republic of Iran.

The exact determination of gene defect for thalassemia carriers is essential for premarital screening. It will also help the genetic counselor to advise the family accordingly. Among 142 blood samples from PND (prenatal diagnosis) center of Pasteur Institute of Iran which had exact mutation on beta globin gene, we performed simultaneously some molecular experiences such as Multiplex gap-PCR, ARMS-PCR and sequencing for detecting probable mutation on their alpha globin gene cluster. All samples (142) had 25 different mutations on their beta globin gene, but just one mutation on alpha globin gene in 16 samples successively identified. 15 samples were heterozygote and one sample was homozygote for deletional alpha mutation named-3.7. All these 16 samples had 8 different mutations on their beta globin gene, on which 7 samples were IVSII-I and 3 samples were IVSI-5 and simultaneously all 16 samples had -3.7mutation on their alpha globin gene cluster. Through these 16 samples, the descriptive statistics of the mean value of HbA2, were 4.2 1.0, and for MCV: 67.35.8, and for MCH: 20.92.7. Type and frequency of mutations in population with high prevalence of hemoglobin disorder is very variable. In the several publications from our geographical region, it is obvious that IVSII-I is the most prevalent mutation in the beta thalassemia patients and the most prevalent mutation among alpha thalassemia carriers is -3.7. Co-presence of both mutation in globin genes in chromosome 16 and chromosome11, is distinctly -3.7 with IVSII-I. P01.61 Feasibility of Preconception screening for thalassaemia in indonesia: Exploring the Opinion of Javanese mothers
C. G. Widayanti1, A. Ediati1,2, M. Tamam3, S. M. H. Faradz1,2, E. A. Sistermans4, M. C. Cornel4,5, A. C. Plass4,5; 1 Diponegoro University, Semarang, Indonesia, 2Center for Biomedical Research (CEBIOR), Semarang, Indonesia, 3Dr Kariadi Hospital Diponegoro University, Semarang, Indonesia, 4VU University medical center, Amsterdam, Netherlands, 5 The EMGO-institute for Health and Care research, Amsterdam, Netherlands.

We have investigated the clinical usefulness of the Activated protein C resistance (APCR)/factor V Leiden mutation (FVL) test by sending out questionnaires to all Norwegian physicians who ordered these tests from our publicly funded service laboratory during a three months period, and 70% (267/383) responded. Indications for testing, patient follow-up, the use of APCR versus FVL tests, and differences in practice between hospital doctors and GPs were examined. We found that 46% of the APCR/FVL tests were predictive tests, ordered for risk assessment in healthy individuals with no previous history of VTE. Among these, 42% of the tests were taken on the initiative of the patient, and 24% were screening tests before prescription of oral contraceptives. Fifty-four per cent of the tests were classified as diagnostic, among which 42% were ordered due to a previous history of VTE, and 22% due to a history of brain stroke or myocardial infarction. The prevalence of FVL heterozygotes was not significantly different between the predictive and diagnostic test groups, 26% and 20%, respectively. Only the predictive tests influenced patient follow-up. Here, physicians advice to patients depended on the test result. In general, the clinical usefulness of APCR/FVL testing was low. Many tests were performed on unsubstantiated or vague indications. Furthermore, normal test results led to unwarranted refrain from giving advice about antithrombotic measures, to the potential harm of the patient. P01.63 How do clinical geneticists-in-training spend their working hours?
B. R. Diness; Department of Clinical Genetics, Juliane Marie Centret, Rigshospitalet, Copenhagen OE, Denmark.

Background: Thalassaemia has become a major health problem in Indonesia. It has been estimated that about 10% of the population carries the mutated gene. However, there are no formal recommendations for thalassaemia screening. This study aimed to explore awareness of thalassemia, and attitude regarding carrier testing among Javanese mothers. Methods: A Quantitative questionnaire was applied cross sectional, using constructs of the Theory of Planned Behavior. Results: 180 mothers were invited: 74 had a child affected with thalassemia (RR=100%). Both, attitude towards receiving information about thalassemia (M=4.08), and attitude towards carrier testing (M=4.09) were high. Awareness was low: mothers whose children were not affected hardly ever heard of thalassemia (N=106; M=1.58), whereas mothers with an affected child showed high interest in carrier testing (N=74; M=4.02). Respondents did not perceived control over carrier testing (M=3.02), and feared being discriminated against or stigma-

Aim: To determine how clinical geneticists in training spend their time and to compare this to doctors in other specialties. Method: Time-motion study. A clinical geneticist-in-training recorded her activities prompted by an alarm clock at ten minutes intervals during five consecutive workdays. Activities were categorised as direct patient care, indirect patient care, administrative tasks, conference participation, external professional communication of laboratory results and test options, research, break, and other. Results: 235 observations were recorded. The proportion of time spent on different categories of work is shown in table 1. Approximately half of the time was used on patient care, but less than a third of this time was spent directly interacting with patients. Discussion: Studies of doctors-in-training are sparse, but one large study from Australia documenting the activities of doctors-in-training in internal medicine reported similar time fractions spent on direct and indirect patient. Other studies of how doctors in various specialties spend their time report with few exceptions that less than half of the time is spent on patient care and that most patient care takes place without the presence of the patient (charting, reviewing test results, etc.). Conclusion: The workday of doctors-in-training in clinical genetics may not differ as much from other medical specialties as commonly perceived.

Clinical genetics and Dysmorphology

Table 1
category of activity Direct patient care Indirect patient care Administrative tasks Conference participation External communication Research/project Break Other Proportion of time spent 14 % 32 % 11 % 8% 17 % 4% 6% 8%

7
results in the affected children are similar to those previously reported with evidence of increased growth in the pre-puberal age and an average incidence of congenital malformation and health needs. Mean age at first word was 15 months, showing a slight delay in language skills which tended to improve by the time they reached school age. Parental responses to the interview demonstrated residual anxiety but with a satisfactory adaptation to and a positive recall of the prenatal counselling session. We believe that an integrated approach to prenatal counselling is the best way to manage the anxiety and falsely imagined consequences which parents feel after being told that their fetus bears such a genetic abnormality. P01.66 Influence of associations of patients in rare diseases (Williams syndrome and smith magenis syndrome) in clinical Research. Research centers of reference

P01.64 Professional training for the new era of genetic medicine

A. C. Davies1,2, K. Dack1,2, M. Leech1,2, D. Donnai1,2, H. R. Middleton-Price1,2; 1 Nowgen A Centre for Genetics in Healthcare, Manchester, United Kingdom, 2 Central Manchester University Hospitals NHS Foundation Trust, Manchester, United Kingdom.

Genetics and genomics are making a dynamic and expanding contribution to mainstream healthcare, for example: the increasing use of pharmacogenetics and companion diagnostic testing to personalise medicine. In addition, genetic knowledge is increasing at such a pace that the rate of data generation is at risk of overtaking the capacity for its interpretation. Nowgen, a Centre for Genetics in Healthcare, is at the forefront of training provision in the arena of genetic medicine. In collaboration with our colleagues in the Central Manchester University Hospitals NHS Foundation Trust, The University of Manchester and the National Genetics Reference Laboratory, Manchester, we have developed a portfolio of training courses for health professionals, industry professionals and research academics in response to expressed needs for training in these diverse areas. Here we demonstrate Nowgens role in training provision, including the following areas: Bioinformatics for clinicians and scientists working in genetic medicine, to help develop skills interpreting genetic data; Genetics for research ethics committees, to provide a tool-kit to support committee members in their ethical review of genetic research studies; Oncology-based pharmacogenetics, which aims to provide an introduction to this area, including the regulatory issues, effect on clinical trial design, diagnostic testing and data interpretation; and Molecular genetics for cytogeneticists and genetic counsellors, which aims to provide attendees with an understanding of molecular techniques and analytical methods used in molecular genetics laboratories. All courses have been developed in close consultation with senior representatives from the relevant professional groups. P01.65 clinical characterization and analysis of temperamental traits and impact on their parents in 42 italian young girl with triple X syndrome.

M. Fernndez-Prieto1, E. Garayzbal2, M. Lens1, A. Sampaio3, M. Buceta4, . Carracedo5; 1 Fundacin Pblica Galega de Medicina Xenmica, Santiago de Compostela, Spain, 2Universidad Autnoma de Madrid, Madrid, Spain, 3Universidade do Minho, Department of Psychology, Braga, Portugal, 4Universidade de Santiago de Compostela, Department of Evolutive Psychology, Santiago de Compostela, Spain, 5Group for Genomic Medicine of University of Santiago de Compostela, Santiago de Compostela, Spain.

E. Folliero1, D. Quagliarini2, U. Cavallari3, B. Gentilin1, P. Castorina1, F. Forzano4, S. Forzano5, E. Grosso6, S. Gattone7, F. Faravelli8, L. Gargantini9, F. Lalatta1; 1 UOS Dipartimentale di Genetica Medica, Milano, Italy, 2UO Ostetricia e Ginecologia, Milano, Italy, 3Centro Malattie rare cardiologiche, UO cardiologia AO Sacco, Milano, Italy, 4SSD Genetica medica E.O. Ospedali Galliera Genova, Genova, Italy, 5Genetica Medica AOU S. Giovanni Battista and Dipartimento di Pediatria, Universit di Torino, Torino, Italy, 6Genetica Medica, AOU S. Giovanni Battista, Torino, Italy, 7Genetica Medica, Genova, Italy, 8SSD di Genetica Medica, EO Ospedali Galliera, Genova, Italy, 9UO Pediatria A.O Treviglio, Treviglio, Italy.

In rare genetic diseases research it is a critical issue to build national and international networks of study. Platforms providing the necessary logistics to carry out the proposed work of researchers and clinicians are also needed, due to geographic dispersion. This usually occurs because of the low incidence of rare diseases. Logistic platforms integrating information and centralized federations of rare diseases allow associations concerned to coordinate their activities, ranging from research to therapeutic activities, including leisure and family entertainment. Also, they allow to overcome the barrier of geographical dispersion. In this context, the Association of Williams Syndrome of Spain has a long tradition in the collaboration with researchers. By other hand, the Spanish Association of Smith Magenis has been recently created. Both associations have joined together to promote research, integrating professionals who study these diseases, in order to develop different projects and clinical programs. They found strategic support in the Regional Center for Rare Diseases (CREER) that allows cooperation among associationism and research. The Spanish Federation for Rare Diseases also supported them, and also another specific centers of genetics as the Galician Public Foundation of Genomic Medicine did so. This allows to establish a real national internal network of detection and early diagnosis of these diseases, facilitating research. It also helps to develop a better understanding of the neurocognitive, clinician and educational profiles affected and to publish specific materials for different diseases. The main objective of this paper is to present the existing networks in Spain studying rare diseases. P01.67 Role of genetic family testing in Wilson disease.

N. Mocanu, S. Groppa; National Center of Reproductive Health and Medical Genetics, Chisinau, Moldova, Republic of.

We report clinical and temperament evaluation data in a large Italian cohort of young girls with triple X syndrome whose diagnosis was made prenatally between 1998 and 2006 in three Italian Centres. At initial evaluation reproductive and medical histories were collected. Clinical assessment of the child was performed by a clinical geneticist and included a detailed personal history, physical evaluation and auxological measurements. To analyze how parents coped with specific events in the prenatal and postnatal periods, we conducted an interview which included 35 specific questions designed to elicit retrospective judgements on prenatal communication, present and future worries, needs and expectations. In a subset of probands we also administered the formal Italian Temperament Questionnaire assessment test which investigates adaptation, general environment and socialization. This test also assesses the emotional component of temperament. Clinical

Wilsons disease (WD) represents a severe hereditary disorder that predominantly affects liver and brain which in case of late revealing and inefficient treatment or its absence leads to severe disabilities, demands considerable financial expenses and even might be fatal. Affected individuals may require health, financial and personal care for 10-15 years. This care is frequently undertaken by family members. We investigated Moldavian families with WD for identification of mutations at members of families for the purpose of carrying out of preliminary presymptomatic therapy. We analyzed 34 Moldavian WD patients and family members who were genetically confirmed. In 8 (23,5 %) cases we revealed diseases in siblings of probands on presymptomatic stage, that given us a chance to carry out required therapy and to avoid appearance of clinical WD symptoms and physical disability in these patients. Disease identification in 23, 5 % presymptomatic patients has a major importance for diagnosis of WD in Moldova. These cases highlight

Clinical genetics and Dysmorphology

the importance of accessible and accurate information about rare diseases. Genetic counseling in WD family and molecular testing has a major practical importance for diagnosis what due to early revealing of patients without clinical symptoms of disease gives a chance to begin appropriate therapy in time, to avoid physical disability and to keep a life in such patients. P01.68 Written information to patients in clinical genetics: whats the impact?

Pharmacy, Craiova, Romania, 5University Hospital San Carlos, Madrid, Spain.

C. Cassini1, C. Thauvin-Robinet1, F. Coron1, S. Vinault2, C. Binquet2, S. Mercier3, N. Laguette3, L. Faivre1; 1 Centre de Gntique, Dijon, France, 2Centre dInvestigations Clinique pidmiologie Clinique, CHU Bocage, Dijon, France, 3Direction des Droits des Patients, de la Qualit et de la Gestion des Risques, CHU Bocage, Dijon, France.

In all countries of the European Union, oral information must be given to the patient. Written information is generally optional, but physicians are tending more and more to send a copy of the clinical report to the patient. In this study, we aimed to evaluate the impact on patients of sending them written information after a clinical consultation in the genetics department. During a three months period, two geneticists and one genetic counselor offered to send each patient a copy of the letter sent to their general practitioners. A questionnaire was sent with this copy. 375 patients were seen, 60% of the questionnaires were sent back. Of these, 99% showed that this practice was considered a good idea, and 82% reported that the letter reflected the clinical aspects well. 72% thought that receiving this letter improved their understanding, and only 9% found the letter confusing. In general, patients found the words understandable (83%) and only 2% were shocked. 58% would have preferred a letter sent specifically to them, and 63% said that they would have asked their general practitioner to show them the letter. Their main motivation for wanting a copy of this letter was to have the information to pass on to other physicians involved in their health in the future, or to have information concerning the family. There were no significant differences depending on the physician, the indication for the clinical consultation, the age or gender of the patients, or their level of general understanding. J01.1 Early Diagnosis and screening of congenital Heart Anomalies

A mutation of the fibrillin-1 gene gives an inherited multisystemic connective-tissue disease, called Marfan syndrome. It is characterized by a wide range of clinical manifestations. The major mortality generators are the cardiovascular manifestations: annuloaortic ectasia, aortic dissection, aortic aneurysm, pulmonary artery dilatation and mitral valve prolapse. Common musculoskeletal manifestations include: scoliosis, pectus excavatum and carinatum, arachnodactyly and acetabular protrusion. Dural ectasia is a characteristic nervous system affection. In some patients there is also pulmonary and ocular involvement. To improve the life expectancy and quality, an early identification and treatment of these clinical aspects is a must. Geneticists play a key role in Marfan syndrome diagnosis. For a better patient care, a comprehensive diagnostic is driven by a prenatal genetic counseling and the knowledge about the various manifestations of Marfan syndrome. J01.3 Genetic screening in Europe

P. Javaher1, E. Nyoungui1, H. Kriinen2, U. Kristoffersson3, I. Nippert4, J. Sequeiros5, J. Schmidtke1; 1 Department of Human Genetics, Hannover Medical School, Hannover, Germany, 2Department of Medical genetics, University of Turku, Turku, Finland and National Institute for Health and Welfare, Helsinki, Finland, 3Department of Clinical Genetics, University Hospital, Lund, Sweden, 4Department of Human Genetics, Universitaetsklinikum Muenster, Muenster, Germany, 5UnIGENe and CGPP, IBMC Institute for Molecular and Cell Biology and ICBAS, University Porto, Porto, Portugal.

M. Taghizadeh, S. Dastgiri, M. Heidarzadeh; Department of Community and Family Medicine, Tabriz, Islamic Republic of Iran.

There are many cases of congenital heart anomalies missed at the time of birth. Literature demonstrates that CH anomalies are the second most common birth defect in many countries. Despite this fact, our previous study showed that the prevalence of CH anomalies is the fifth most common one indicating that many of these defects are not diagnosed at the time of birth. The aim of this study was to estimate the missing frequency of CH anomalies at the time of birth. The population of the study was 185650 births (183579 live births and 2071 still births) in the northwest of Iran covered by Tabriz Registry of Congenital Anomalies (TRoCA). A total of 451 children with CH anomalies were studied in this region from 2000 to 2009. The expected prevalence of CH anomalies at birth was estimated to be 24.2 per 10000 births (CI 95%: 22.1-26.5) while a prevalence of 9.2 per 10000 births (CI 95%: 7.8-10.6) was observed at the same time/place. This indicated that 59.1 percent of children with CH anomalies were not identified at birth (P<0.05). the same proportion increased by 13 percent over the study from 2000 to 2009 (P>0.1). The result indicated that a remarkable frequency of CH anomalies was not diagnosed at birth because there was no pediatric cardiologist available at birth in the gynecology and obstetrics wards. This may necessitate the presence of pediatric cardiologist at the time of delivery or soon after birth for early diagnosis and screening of CH anomalies. J01.2 marfan syndrome in children

Background: Genetic screening has been defined as any kind of test performed systematically for the early detection or exclusion of a genetic disease, genetic predisposition or resistance to a disease, or to determine whether a person carries a gene variant, that may produce disease in his or her offspring. Methods: The data were collected from answers of experts via a selfdesigned questionnaire, which addressed the conditions screened, screening methods, organisational aspects of screening programmes, and conditions screened in prenatal, population-based carrier, and cascade screening programmes, as well as data from websites of national screening authorities and societies and several organisations dealing with neonatal screening issues. Results and conclusion: The current (2006-2008) status of genetic screening and the organisation of genetic screening programmes in selected European countries is presented in this survey as a background for future attempts of harmonizing standards and procedures of genetic screening, an explicit aim of the European Network of Excellence, EuroGentest (www.eurogentest.org).

P02 clinical genetics and Dysmorphology

P02.001 interstitial deletion of 10p12-p11: A novel micro-deletion syndrome associated with mental retardation and a distinct phenotype
C. Wentzel; Department of Genetics and Pathology, Uppsala University, Uppsala, Sweden.

R. Pakai1, D. Iacob2, R. E. Iacob3, P. Stancu4, M. Puiu2, M. Manescu5; 1 Louis urcanu Childrens Emergency Hospital, Timisoara, Romania, 2 University of Medicine & Pharmacy, Victor Babe, Timisoara, Romania, 3 Pediatric Surgery, SCJU, Arad, Romania, 4University of Medicine and

The ability to detect cryptic unbalanced rearrangements in patients with syndromic mental retardation has improved considerably with the clinical implementation of genomic microarrays. Here we report on the molecular karyotyping of three unrelated patients with partially overlapping microdeletions at chromosome 10p11.23-p12.1 ranging from 0.99 Mb to 4.03 Mb in size. The smallest region of overlap (SMO) between the deletions is 400 kb. Yet another patient has previously been described with a 10 Mb deletion, partially overlapping with our three patients {Shahdadpuri, 2008 #1}. All four patients have a developmental delay and growth retardation. Apart from the SMO, the genomic region could be divided into three separate intervals when related to phenotypic expression and symptoms. Together with the previously reported patient our study suggests that the deletions may represent a novel micro-deletion syndrome with a distinct clinical expression associated with haploinsufficiency for genes in this region. Reference Shahdadpuri R, de Vries B, Pfundt R, de Leeuw N and Reardon W. American Journal of Medical Genetics Part A 146A: 233-237 (2008)

Clinical genetics and Dysmorphology

P02.002 Deletion of 13q32.3-34: A genotype-phenotype correlation of three cases

Amiens University hospital, amiens, France, 6department of genetics, Nantes University hospital, Nantes, France, 7department of genetics, Lille University hospital, Lille, France.

J. Davidsson1, H. gerstam1, M. reberg2, M. Soller1; 1 Department of Clinical Genetics, University and Regional Laboratories, Skne University Hospital, Lund, Sweden, 2Department of Pediatrics, Kalmar Hospital, Kalmar, Sweden.

Background: Partial deletions of chromosome 13 are rare. Based on their clinical and cytogenetic features patients have been categorized into three groups: 1) deletions proximal to 13q32, 2) deletions involving 13q32; and 3) deletions of 13q33-34. Of these, group 2 has the most severe phenotype, including, among others, dysmorphology of the brain, eyes, gastrointestinal tract and distal limbs. Methods: We have collected clinical data and performed molecular cytogenetic mapping of three cases with de novo deletions of 13q32.334, using metaphase FISH and genome-wide array CGH. Results: In case 1, FISH analysis mapped the deletion to range from 99.53 Mb to pter on chromosome 13. The clinical symptoms included, among others, developmental delay, an intracranial dural arteriovenous fistula and low levels of clotting factors 7 and 10. In case 2, FISH analysis mapped the deletion to span from 107.74 Mb to pter. The clinical symptoms included, among others, developmental delay, microcephaly, hypertelorism and low levels of clotting factors 7 and 10. In case 3, array CGH mapped the deletion to range from 101.07 Mb to pter. The clinical symptoms included, among others, developmental delay, microcephaly and hypotonia. Conclusions: We present three cases with 13q32.3-34 deletions, characterized at the molecular cytogenetic level. In the deleted regions, several candidate genes responsible for the diverse phenotypes of the patients could be identified. Characterization of additional patients harboring similar distal deletions might lead to better, and clinically more useful, description of the 13q deletion syndrome. P02.003 Narrowing down the minimal critical deletion region of recurrent 16p11.2-p12.2 microdeletion syndrome

Duplication of the critical Rubinstein Taybi deletion region on the chromosome 16p13.3 have recently been proposed to be a cause of a recognizable syndrome, characterized by normal to moderately retarded mental development, normal growth, mild arthrogryposis, mild abnormalities of the hands, characteristic facial features, and occasional anomalies of the heart. Up to date, only 13 patients with an interstitial duplication of the 16p13.3 region encompassing the CREBBP gene have been reported. We report 4 new cases with such 16p13.3 microduplication. They present mental retardation (4/4), normal growth (4/4), mild arthrogryposis anomalies (2/4), and no anomalies of the heart. Remarkable facial features such as microretrognathia (4/4), broad nose (4/4), dolicocephaly (2/4) were noted, and also frequent eye involvement (4/4) with upslanting eyes with narrow palpebral fissures (4/4), ptosis (3/4), bilateral epicanthi (3/4). Minor anomalies of the extremities- hands and feet- were always found (4/4), with frequent involvement of the thumb: proximally adducted thumbs (3/4), adducted thumbs (2/4). Several other occasional findings reported before were also noted : cleft palate (2/4)/ bifid uvula (1/4), inguinal hernia (2/4), hyperconvex nails (2/4) and hammer halluces (1/4)/short halluces (1/4). One patient also presents retinal hypogmentation. A detailed clinical description of the 4 patients with review of the literature will be made, confirming the recognizable microduplication syndrome, and helping to delineate the associated phenotype. P02.005 microduplication of 17p13.3 including YWHAE in a patient with strongly increased nuchal translucency as a fetus, and postnatal abnormal phenotype.
D. Svaneby1, J. Graakjaer1, G. Dybmose2, C. Fagerberg1; 1 Department of clinical Genetics, 7100 Vejle, Denmark, 2Department of pediatrics, Esbjerg, Denmark.

K. M. Roetzer1, A. C. Obenauf1, B. Plecko-Startinig2, M. Brunner-Krainz2, M. R. Speicher1; 1 Institute of Human Genetics, Graz, Austria, 2Department of Pediatrics, Graz, Austria.

Microdeletions in16p11.2-p12.2 were recently described in a number of patients exhibiting mental retardation, speech delay, developmental delay, hypotonia, dysmorphic features, and recurrent infections. Yet, due to the size of the deletions (~ 8Mb) and the number of genes contained it was difficult to link the deleted genes to the phenotype. Here we report the case of an 11-year-old girl with muscular hypotonia, speech delay, ptosis, recurrent infections and dysmorphic features similar to those reported in patients with microdeletions in 16p11.2-p12.2. A muscle biopsy at the age of 1 year showed unspecific changes, whereas the altered structure of the mitochondria was most striking. However, biochemical investigations for mitochondriopathies were inconclusive. Array-CGH analysis revealed a 205kb microdeletion in 16p11.2, overlapping the deleted regions in the 6 patients recently reported. The microdeletion in our patient was inherited from the mother, who following anamnesis and physical examination, also exhibited mild muscular hypotonia, ptosis, and recurrent ear infections. The microdeletion contains only three disease causing genes: TUFM, ATP2A1 and CD19. We hypothesize that the muscular hypotonia could be due to a deletion of ATP2A1 and the recurrent infections might be associated with CD19 deficiency. This theory is further supported by the observation that only one patient reported so far did not exhibit muscular hypotonia, and this was also the only patient with a microdeletion not including ATP2A1. However, further investigations to confirm the central role of ATP2A1 and CD19 in the phenotypic presentation of patients with 16p11.2-p12.2 microdeletions are necessary. P02.004 16p13.3 microduplication syndrome : report of 4 cases and futher phenotype delineation

Background: Deletion of 17p13.3 causes Miller-Dieker syndrome. Duplications in this region have been described with a variable phenotype depending on which genes are included. The genes PAFAH1B1, encoding LIS1, and YWHAE seem to be important for the genotypephenotype correlation. Duplications including the YWHAE gene but not PAFAH1B1 have been described in a few patients, with common features being facial dysmorphism, macrosomia, and mild developmental delay. Individuals with duplications including PAFAH1B1 have no particularly dysmorphic features, but more severe developmental delay, and brain defects. Case: Our patient is a boy, 2 years old. The prenatal ultrasound scan showed an increased nuchal translucency of 7,5 mm, no malformations were seen. A normal male karyotype was found. At the age of 2 years the patient showed increased growth and psychomotor retardation. He was hypotonic, had a bad temper, and expressive and impressive language delay. Facial dysmorphism included epicanthus, low set ears, smooth philtrum, thin upper lip, and short nose. Array-CGH showed a de novo 1.02 MB duplication on chromosome 17p13.3. The duplication included the YWHAE gene, but not the PAFAH1B1 gene. Conclusion: A few cases of duplication 17p13.3 have been described with variable phenotype depending on whether YWHAE or PAFAH1B1 is involved. We here describe a patient with duplication including YWHAE with very similar features to previous cases, but also with an increased nuchal translucency, thus extending the phenotypic spectrum. It can be considered in pregnancies with very high nuchal translucency combined with normal karyotype to investigate for duplications in 17p13.3. P02.006** the 17q21.31 microdeletion syndrome

D. A. Koolen, B. B. A. de Vries; Radboud University Nijmegen Medical Centre, Nijmegen, Netherlands.

B. Demeer1, G. Morin1, S. Julia2, D. Martin-Coignard3, G. Plessis4, F. Amram1, A. Receveur5, H. copin5, G. Bourrouillou2, C. Le Caignec6, J. Andrieux7, M. Mathieu1; 1 department of genetics, Amiens University hospital, amiens, France, 2 department of genetics, Toulouse University hospital, Toulouse, France, 3 department of genetics, Le Mans hospital, Le Mans, France, 4department of genetics, Caen University hospital, Caen, France, 5department of cytogenetics,

The 17q21.31 microdeletion syndrome is a novel genomic disorder with a prevalence of around 1/16,000. We present an update of the delineation of the phenotype based on the clinical description of 47 individuals with the recurrent 17q21.31 deletion and call for an international collaboration to collect information on this emerging syndrome (www.17q21.com). In our cohort of patients the most consistent features are hypotonia and mild to severe global psychomotor developmental delay. The facial dysmorphisms include a high/broad forehead,

Clinical genetics and Dysmorphology

long face, upward slanting palpebral fissures, epicanthic folds, an abnormally shaped nose, large prominent ears and an everted lower lip. Abnormal hair pigmentation and texture is also frequently present. The facial phenotype may evolve with age, leading to coarsening and elongation of the face. Short stature, pectus excavatum, spine anomalies, dislocation of the hip(s), long slender fingers and slender lower limbs, and positional deformities of the hand/feet are also reported. A history of epilepsy is noted in ~50% of all cases. Other serious abnormalities include atrial and ventricular septal defects, kidney/ urologic anomalies, and cryptorchidism. Our data show that the 17q21.31 microdeletion syndrome is a clinically well recognizable genomic disorder. Longterm studies, however, are required to further define the syndrome and to determine the prognosis in adult patients. Acknowledgements: Many thanks to our collaborators, for providing DNA samples and clinical and/or molecular data. P02.007 Loss of cytochrome P450 17A1 Protein Expression in a 17-Hydroxylase/17,20-Lyase-Deficient 46,XY Female Caused by two Novel mutations in the CYP17A1 Gene
S. Kofman-Alfaro1, N. Njera1, I. Palma1, R. Pea2, G. Queipo1; 1 Hospital General de Mexico-Facultad de Medicina UNAM, Mexico City, Mexico, 2Hospital Infantil de Mxico Federico Gmez, Mexico City, Mexico.

0
gion at 1p36.31 for ,dysgenesis of the corpus callosum and epilepsy. P02.009 severe Lysosomal Lipid storage Disease of the Liver in monosomy1p36: New Presentation Extending the Phenotype
M. Haimi1, T. Iancu2, L. Shaffer3, A. Lerner4; 1 Childrens Health Center, Clalit Health Services, Haifa, Israel, 2Pediatric Research and Electron Microscopy Unit, Technion-Rappaport Faculty of Medicine, Haifa, Israel, 3Health Research and Education Center, Washington State University & Signature Genomic Laboratories, Spokane, WA, United States, 4Pediatric Gastroenterology and Nutrition Unit, Carmel Medical Center, Haifa, Israel.

17-hydroxylase deficiency (17OHD) is a rare form of congenital adrenal hyperplasia (CAH) caused by mutations in the CYP17A1 gene. This condition shows considerable clinical and biochemical variation. Molecular characterization of novel mutations in the CYP17A1 gene and detailed study of their structural, enzymatic, and clinical consequences are required to fully understand enzyme behavior. Here, we present the first molecular characterization of two novel mutations in CYP17A1 in a 15-year-old female Mexican mestizo 46,XY female with primary amenorrhea and lack of pubertal development, and severe hypertension that manifested only after surgery. A complete clinical and biochemical evaluation was compatible with 17OHD. Structural anomalies in the CYP17A1 gene were discovered by direct automated sequencing, which revealed a novel compound heterozygous K110X/R362H mutation that leads to a complete lack of enzyme activity. Immunohistochemical analyses performed to determine protein expression and localization showed that cytochrome P450 17A1 was completely absent in the patients testicular tissue. Studies of novel mutations, such as those described here, provide important information that allows us to better understand the effect of a given mutation on enzyme function and to observe the impact of the mutation on clinical phenotype. P02.008 Three Interrupted Deletions at 1p36 in a Patient with Lateral Ventricular Enlargement, Epilepsy and corpus callosum Abnormality
S. Cingz1, A. nalp2, S. Betinolu1, S. Kurul3, Z. Tmer4; 1 Department of Medical Biology and Genetic, School of Medicine, Dokuz Eyll University, zmir, Turkey, 22Department of Pediatrics, Division of Pediatric Neurology, Behcet Uz Child Disease and Pediatric Surgery Training and Research Hospital, zmir, Turkey, 33Department of Pediatrics, Division of Child Neurology, Dokuz Eyll University School of Medicine, zmir, Turkey, 4Kennedy Center, Glostrup, Denmark.

Background /Aim : Monosomy 1p36 has been increasingly recognized as a distinct chromosome deletion syndrome in the past few years. Monosomy 1p36 is mostly associated with severe mental retardation, developmental delay, behavioral difficulties and self-injury. There are several distinct dysmorphic features, including large anterior fontanels, microcephaly, brachycephaly, deep-set eyes, flat nose, nasal bridge and pointed chin. In contrast to the classical phenotype, several children with a 1p deletion have had overgrowth and hyperphagia with a clinical presentation similar to Prader -Willi syndrome (PWS). Methods: Here we describe an 11-year-old girl with 1p36 deletion demonstrating the classical dysmorphological features, having developed an uncontrolled voracious appetite and severe truncal obesity. Gastroenterological evaluation revealed elevated liver enzymes. Liver biopsy disclosed severe fatty liver: in addition to medium-size triglyceride droplets, hepatocytes showed excessive lipofuscin accumulation. A most unusual feature was the presence of frequent, extremely large lipolysosomes, never previously reported in this condition. Results: Oligonucleotide-based microarray analysis was performed using a 105K-feature whole-genome microarray. It showed copy-number loss of 177 oligonucleotide probes from the short arm of chromosome 1 at 1p36.33p36.32, approximately 2.2 Mb in size. Conclusions: We suggest that the chromosome segment 1p36.3336.32 harbour a critical region for the manifestation of obesity and hyperphagia. We also suggest that monosomy 1p36 syndrome should be considered in patients with hypotonia, developmental delay, obesity, hyperphagia, behavioral disorders, learning difficulties and a negative genetic test for PWS, even in the absence of the striking facial features of the syndrome. P02.010 BAc array analysis detects microdeletions on chromosome region 20p12.1 in two unrelated individuals with mR/ mcA syndrome

L. Grozdanova1, R. Stoeva2,3, J. Vermeesch3, J. P. Fryns3, B. Dimitrov3, R. Thoelen3, M. Stefanova4; 1 Department of Medical Genetics, University Hospital, Plovdiv, Bulgaria, 2 Department of Pediatrics and Medical Genetics, Medical University, Plovdiv, Bulgaria, 3Center for Human Genetics, Katholieke Universiteit Leuven, Leuven, Belgium, 4Department of Clinical Genetics, Sahlgrenska University Hospital, Gothenburg, Sweden.

Monosomy 1p36 is a frequent terminal microdeletion syndrome characterized by distinct craniofacial features, developmental delay, mental retardation, hypotonia, seizures, visual, auditory, structural brain and cardiovascular malformations. The 1p36 deletion syndrome is likely to be associated with haploinsufficiency of contiguous genes. In previous studies KCNAB29 (voltagegated potassium channel -subunit gene) and GABRD (the human -aminobutyricacid A receptor delta-subunit gene) were suggested as candidate genes for epilepsy. Genes within the 2.2 Mb smallest region of overlap have also been suggested as potential candidate genes for periventricular nodular heterotopias (PH). Here we report a case with three consecutive interstitial deletions at 1p36.22-p36.23, 1p36.31 and 1p36.32array CGH. All the three deletions were within the common deletion region (1pter-p36.23) of classical monosomy 1p36 patiens.The present case has mental retardation, developmental delay, dysgenesis of the corpus callosum, epilepsy, lateral ventricular enlargement and facial dysmorphism. In this study we comparedthe phenotypes and the deletion sizes of the present case with previously published cases. Our study suggests a new critical re-

One patient with a de novo deletion spanning a 250kb region of 20p12.1 and clinical features of Kabuki syndrome was described previously (J Med Genet 2007 44:562-569). Here we report a detailed clinical and molecular investigation of two unrelated individuals, a 15-years-old and a 7-years-old boys with respectively paternally and maternally inherited overlapping deletions of 20p12.1, detected by BAC array CGH at 1 Mb resolution. Clinically they both presented with characteristic dysmorphic features, dental abnormalities, gum hypertrophy, brain malformations and severe mental retardation. Common dysmorphism include sloping forehead, arched eyebrows, long palpebral fissures, long eyelashes with unusual growth pattern at the lower eyelid, short philtrum, open-mouth appearance, high palate, posterior rotated ears with massive earlobes. Failure to thrive during infancy, periorbital and generalized oedema, hallux valgus were noted. Additionally, the 15-years-old patient presented with nasolacrimal duct stenosis, single palmar crease, thenar hypoplasia, cubitus valgus, pes planus, stereotypic movements. The 7-years-old boy developed a short stature (SD-4.7) and microcephaly (SD-2.5). He showed epilepsy, retinal pigment changes, cataract, myopia, bilateral congenital hip luxation and cryptorchid testes. A deletion of the BAC clone RP5-855L24 (14609853-14755918 bp, HG18) was detected in all cells with the 15-years-old patient and his father and as well as with the other and his mother.

Clinical genetics and Dysmorphology

The similar clinical presentation in both individuals suggests the causative effect of the aberration. Epigenetic factors could be potential reasons for the healthy parents carrier. To our knowledge these are the first two cases reported with microdeletions 20p12.1 and inherited from the healthy parent. P02.011** Drosophila as a model organism to establish a genotype-phenotype correlation in 2q23.1 deletion syndrome
B. W. M. van Bon, J. Kramer, A. Schenck, B. B. A. de Vries; Radboud University Nijmegen Medical Centre, Nijmegen, Netherlands. Universitt Dresden, Dresden, Germany.

1
Subtelomeric deletions of chromosome 6q have emerged as a characteristic microdeletion syndrome, characterized by mental retardation, seizures, mild dysmorphic features, and brain anomalies. The latter most commonly include enlarged ventricles or hydrocephalus and aplasia/hypoplasia of the corpus callosum. Thus far, aplasia of the olfactory bulbs and tracts has only been reported as autopsy findings in two patients with a ring chromosome 6 and in one patient with a 6q23-deletion. We report on two patients with a de novo subtelomeric rearrangement involving the long arm of chromosome 6 (6q27) detected by molecular karyotyping using Array-CGH analysis. Both patients had aplasia of the olfactory bulbs and hydrocephalus due to aqueduct stenosis as well as additional brain malformations. The first patient presented with global developmental impairment, facial dysmorphism, and had an unbalanced reciprocal translocation of chromosome 3 and 6, resulting in a 6q27 deletion and an additional 3q29 duplication. The second patient presented with complete anosmia as the only clinical symptom and showed no signs of delayed physical or mental development. Thus we conclude that there is considerable variability in the phenotypic spectrum of the disorder and that the degree of developmental impairment does not correlate with the extent of brain findings. Therefore the 6q Terminal Deletion Syndrome might actually be underdiagnosed as it can manifest by anosmia only. Moreover, since olfactory bulb aplasia seems to belong to the phenotypic spectrum of the 6q terminal deletion syndrome we suggest screening these patients for this malformation by high resolution brain imaging. P02.014 A case report of Achondroplasia (AD inheritance) in a family with consanguineous marriage( both parents are not affected).

In general, patients with a 2q23.1 microdeletion are mentally retarded and present with speech delay, short stature, microcephaly, seizures, coarse facies, sleep disturbances and behavioral problems. These features may lead to the initial clinical impression of Angelman, Rett and/or Smith-Magenis syndromes. The overlapping 2q23.1 deletion region in all reported patients comprises one gene, MBD5, which is a member of the methyl CpG-binding domain protein family. Another gene in this region, EPC2, is also deleted in the majority of patients, who appear to have a more severe phenotype than those with a deletion of MBD5 only. EPC2 is a member of the polycomb protein family, involved in heterochromatin formation. Several genes with a role in modulation of chromatin structures are associated with mental retardation. Therefore, EPC2 might be considered as a mental retardation gene. In the absence of patients with mutations or deletions of EPC2 only, we used Drosophila melanogaster as model organism to elucidate possible pathogenicity of this gene. Ubiquitous knockdown of the fly ortholog epc induces lethality, mainly during pupal stages. Drosophila also offers the possibility of tissue and cell-specific gene ablation. Epc knockdown in wings and eyes, severely disturbes morphogenesis of these organs. These results point to an important function of epc during development and suggest a significant contribution to the pathology of the 2q23.1 microdeletion syndrome. We will now focus on the role of epc in neuronal function and behaviour. This study aims to establish the relevance of fly models in understanding gene-phenotype correlations in microdeletion syndromes. P02.012 situs inversus totalis with 47,XXX; A case report

S. Akbaroghli1,2, M. T. Akbari3; 1 Dr. Akbaroghli Genetic Counselling Center, Tehran, Islamic Republic of Iran, 2 Deputy for Cultural Affairs and Prevention of Iran Welfare Organization, Tehran, Islamic Republic of Iran, 3Tehran Medical Genetics Laboratory, Tehran, Islamic Republic of Iran.

H. Kocak Eker1, M. Ikbal2, A. H. Cebi2, M. Y. ALP2, T. Tos3; 1 Dskap Children Hospital, Dept of Medical Genetics, Ankara, Turkey, 2 Karadeniz Technical University Medical Faculty, Medical Genetics, Trabzon, Turkey, 3Department of Medical Genetics, SB Sami Ulus Womens and Childrens Hospital, Ankara, Turkey.

We report a case of a 1,5 year old female situs inversus totalis with 47,XXX. We think that this is the first case in the literature situs inversus totalis with 47,XXX. She is the first child of the family and has no sister or brother. She was taken to our genetic polyclinic because of her puffy face. She had situs inversus totalis and large VSD on her history. We want to see the karyotype of the patient because she had some abnormalities with her phenotype and we know some cases situs inversus with chromosomal abnormalities. Chromosomal analysis performed on a peripheral blood lymphocyte culture showed a 47,XXX female karyotype. Further studies are needed to explain situs inversus totalis connexion with X chromosome abnormalities. P02.013 Expanding the clinical and neuroradiological phenotype of 6q terminal deletion syndrome: olfactory bulb aplasia and aqueductal stenosis.

An eight years old boy with clinical manifestations including: megalocephaly,short stature ( height = 108 cm , weight =24) ,low nasal bridge, prominent forehead,lumbar lordosis, short tubular bones and radiologic manifestations including: rhizomelic dwarfism, V shape deformity, narrow sacrosciatic notches, small iliac wings,short trident hand. He is the result of a consanguineous marriage ( second cousins).In the molecular study, he has G1138A mutation in FGFR3 gene in heterozygote form. His unaffected parents ( 33 years old woman and 39 years old man) came for preconceptional genetic counseling because of their affected child and consanguineous marriage and they are candidates for prenatal diagnosis because of the probability of gonadal mosaicism. P02.015 Unbalanced translocation t(1;5)(p31.3;q23.2) in a child with congenital malformations and a disorder of glycosylation (cDG) affecting ALG6 gene, and characterized by sNP and cGH arrays

K. Becker1, T. Neuhann1, N. Tyshchenko1, D. Podlesek2, M. Smitka3, S. Tinschert1, G. Hahn4, T. Hummel5, E. Schrock1, J. Gerber6, K. Hackmann1; 1 Institut fr Klinische Genetik, Medizinische Fakultt Carl Gustav Carus, Technische Universitt Dresden, Dresden, Germany, 2Klinik und Poliklinik fr Neurochirurgie, Universittsklinikum Carl Gustav Carus, Technische Universitt Dresden, Dresden, Germany, 3Abteilung fr Neuropdiatrie an der Klinik und Poliklinik fr Kinder- und Jugendmedizin, Universittsklinikum Carl Gustav Carus, Technische Universitt Dresden, Dresden, Germany, 4Abteilung Kinderradiologie am Institut und Poliklinik fr Radiologische Diagnostik, Universittsklinikum Carl Gustav Carus, Technische Universitt Dresden, Dresden, Germany, 5Klinik und Poliklinik fr Hals-Nasen-Ohren-Heilkunde, Universittsklinikum Carl Gustav Carus, Technische Universitt Dresden, Dresden, Germany, 6Abteilung Neuroradiologie am Institut und Poliklinik fr Radiologische Diagnostik, Universittsklinikum Carl Gustav Carus, Technische

M. D. Sanchez-Izquierdo1, E. Bermejo1,2, A. I. Vega3, B. Prez3,4, M. L. Martnez-Fernndez1,4, C. Prez-Cerd3, G. Arriola5, A. Garca5, M. Ugarte3,4, M. L. Martnez-Fras1,6; 1 CIAC, ECEMC, Instituto de Salud Carlos III (ISCIII), Madrid, Spain, 2 Research Institute for Rare Diseases (IIER), ISCIII, Madrid, Spain, 3CEDEM, CBMSO-UAM, Universidad Autnoma de Madrid, Madrid, Spain, 4CIBER de Enfermedades Raras (CIBERER), Spain, 5Servicio de Pediatra, Hospital Universitario de Guadalajara, Guadalajara, Spain, 6Departamento de Farmacologa, Facultad de Medicina, Universidad Complutense, Madrid, Spain.

The genetic study of a 3 year old child with mild psychomotor delay, joint hyperlaxity, talipes valgus, flat frontal hemangioma, total agenesis of the corpus callosum and some dysmorphic features, revealed a de novo translocation detected by a high resolution karyotype and involving chromosome bands 1p31.3 and 5q23.2. For a better characterization of the breakpoint regions, CGH array (244K) and SNP array (500K) analyses were performed, both showing a hemizygous deletion next to ALG6, a gene containing 15 exons and involved in defective glycosylation. The CGH array deletion was 3.5 Mb in size and started at exon 10 of ALG6, while the SNP array deletion was centromeric, and ALG6 was not included in the deleted region. Other genes such

Clinical genetics and Dysmorphology

PGM1 and LEPR where also lost in both arrays analyses. For a better delimitation of the deletion boundaries, PCR amplification of all ALG6 exons was applied to genomic DNA. No product was obtained for exons 11 to 14 when compared to controls, but the amplification of the terminal 3 exon 15 showed a positive result, suggesting an intragenic homozygotic deletion, undetectable by both arrays. An aberrant transcript was obtained using RT-PCR for the detection of ALG6 mRNA. These results suggest a deletion of ALG6 in this patient, affecting both alleles, in a different and non-previously described mode of protein inactivation. Products obtained from the detected rearrangement must be further characterized. Funding for this study provided by: CIBERER (Intra 07/746.1) and ISCIII. Ministerio de Ciencia e Innovacin, Madrid, Spain. P02.016 isolated autosomal recessive hypotrichosis in Pakistan caused by P2RY and LIPH gene mutations
Neurology, Hospital for Sick Children, University of Toronto,, Toronto, ON, Canada.

M. Tariq1,2, A. Azhar2, J. Klar1, I. Ahmad2, S. M. Bakhtiar2, S. M. Baig2, N. Dahl1; 1 Rudbeck Laboratory, Department of Genetics and Pathology, Uppsala University, Uppsala, Sweden, 2National Institute for Biotechnology and Genetic Engineering (NIBGE), Faisalabad, Pakistan.

Autosomal recessive hypotrichosis (LAH) is characterised by diffuse hair loss without associated abnormalities. Typically, hair thinning starts in early childhood and progresses through adolescence. The expression is variable with differences in age of onset and severity of baldness, as well as in hair density and texture. The clinical variation is observed between, as well as within, families segregating hypotrichosis. Recently, the genetic basis of LAH was shown to involve mutations in the P2RY5 gene on chromosome 13q14.3 (OMIM 609239) or the LIPH gene on chromosome 3q27.3 (OMIM 607365). In the present study, we identified eight unrelated consanguineous families from different areas of Pakistan segregating autosomal recessive hypotrichosis. The families were investigated by homozygosity mapping for linkage to P2RY5 and LIPH. Six families showed linkage to the P2RY5 gene and sequencing revealed a single known missense mutation in five families and a novel insertion mutation in one family. The previously known missense mutation was accompanied by a shared haplotype amongst all affected. The remaining two families showed linkage to the LIPH gene and segregated known mutations. The P2RY5 gene encodes a 344 amino acids long G protein-coupled receptor (GPCR). LIPH produces the ligand for P2RY5, acyl-lysophosphatidic acid (LPA). Both genes are highly expressed in the inner root sheaths of the hair follicle and play an essential role in the maintenance of hair growth and texture. From our results, we suggest that a few ancestral mutations cause isolated autosomal recessive hypotrichosis in the Pakistani population. P02.017 carrier frequency for alpha triplication in individuals with normal hematologic indices

Alstrm syndrome is a rare autosomal recessive multisystemic disorder caused by mutations in the ALMS1 gene. The gene is located on 2p13, contains 23 exons and encodes a protein of 4169 amino acids (Collins et al and Hearn et al 2002). Alstrm syndrome is characterized by cone-rod retinal dystrophy causing blindness, sensorineural hearing loss and major organ involvement such as dilated cardiomyopathy, hepatic and renal failure. Childhood obesity, type 2 diabetes, hypertriglyceridemia and hypothyroidism are other common features. Clinical manifestations are highly variable even within family members. We report two siblings with a clinical diagnosis of Alstrm syndrome who presented with early onset rod monochromatism, cardiomyopathy, congenital nystagmus, truncal obesity and mild developmental delay. These 2 sisters are both heterozygotes for a novel pathologic mutation (c.5098A>T) in exon 8 of ALMS1 gene , which causes truncation of the ALMS1 protein. Previous studies suggest that the classical syndrome occurs only in patients with homozygous or compound heterozygous mutations of the ALMS1 gene thus the heterozygosity for this novel mutation does not explain the severity of this disease in our patients. Array-CGH studies for both siblings were normal and MLPA studies of the ALMS1 region are currently pending. Little is known about phenotype-genotype correlation in Alstrm syndrome, although patients with clinical symptoms of Alstrm syndrome were found to have disease-causing mutations in exon 16 (Marshall et al 2007). We hypothesize that rearrangement of the ALMS1 gene in addition to this pathologic mutation is the basis for the severe phenotype of our patients. P02.019 Ambiguous external genitalia in newborns

E. Kiss1, C. Duicu1, C. Banescu2, I. Pascanu3, H. Gozar4; 1 Pediatric Department, University of Medicine and Pharmacy, Tg Mures, Romania, 2Genetic Department, University of Medicine and Pharmacy, Tg Mures, Romania, 3Endocrinology Clinic, University of Medicine and Pharmacy, Tg Mures, Romania, 4Pediatric Surgey Clinic, Emergency County Hospital Tg. Mures, Tg Mures, Romania.

F. Moosavi1,2, A. Amirian1, H. Mirzahoseini1, S. Zeinali1, M. Karimipoor1; 1 Pasteur Institute of Iran, Tehran, Islamic Republic of Iran, 2Science & Research Branch, Azad Islamic University, Tehran, Islamic Republic of Iran.

The 3.7 deletion is the most frequent -globin mutation in Iran, one would then expect that the anti 3.7 triplication is common in Iran as well. the frequency of -thalassemia gene is close to %10 in highly prevalent regions of Iran, so we expect to see frequent co-inheritance of alpha triplication with -thalassemia.Blood samples were selected from individuals with normal hematological indices (MCV>80, MCH>27) (n=153) in Iran population.DNA extraction from peripheral blood leukocytes was performed by salting out method and multiplex PCR was used to detect alpha triplication.Carrier frequency in individuals with normal hematological indices (n=153) was %3.26. This carrier frequency of -triplication is relatively high (3-26) in this study. the coinheritance of this determinant could interfere with genotype-phenotype correlation in minor -thalassemia carriers, and could cause thalassemia intermedia. further characterization by southern blot or MLPA is needed to determine the allele frequency in our population. P02.018 Alstrm syndrome in two sisters.

A newborn infant with ambiguous genitalia is a medical emergency, and the choice of gender must take into account the chromosomal and the gonadal sex, the hormonal influence during fetal life, surgical aspects, and the anatomy of the internal genitalia. A newborn with ambiguous genitalia needs prompt evaluation that will permit gender assignment and detection of life-threatening conditions (salt-losing crisis due to congenital adrenal hyperplasia -CAH). The classification of this disorder in newborns is difficult because similar phenotypes could have several different etiologies. In most cases it is impossible to correlate the etiology of the disorder and the appearance of the external genitalia. Diagnostic criteria applied are similar for all (physical exam, karyotype, investigation of hormones and their derivates, genital ultrasound, radiological examination), but medical and surgical treatment is applied to each patient individually. We present 2 cases of ambiguous genitalia admitted in our department in the last year. RESULTS: In one patients (normal karyotype: 46, XX) the underlying cause of ambiguous genitalia was CAH, while the aethiology of sexual ambiguity in the other case was the chromosomal structural anomalies 46,XY,t(X;21)(p11;q21). CONCLUSION: The etiology of ambiguous genitalia is variable. The physician managing these families could minimize the trauma of having a child with unidentified sex by providing appropriate genetic counseling so that the parents can make an early decision. A proper diagnosis requires a team working: pediatrician, endocrinologist, genetician and pediatric urologist/surgeon. Prenatal diagnosis in at-risk families should be considered and appropriate therapy offered to minimize or prevent genital ambiguity. P02.020 Recurrence of complete arhinia in two siblings with clinical picture of treacher-collins syndrome negative for tcOF1 mutations

S. Jougheh Doust1, M. Nagel2, G. Yoon1,3; 1 Division of Clinical and Metabolic Genetics, Hospital for Sick Children, University of Toronto,, Toronto, ON, Canada, 2Center for Nephrology and Metabolic Disorders, Laboratory for Molecular Diagnostics, Weisswasser, Germany, Weiswasser, Germany, 3Department of Paediatrics, Division of

F. Lalatta1, C. Cesaretti1, B. Gentilin1, E. Folliero1, V. Bianchi1, G. Melloni1, C. Rossi2, C. Meazzini3, M. Bedeschi1, F. Natacci1, R. Brusati4; 1 UOS Dipartimentale di Genetica Medica, Milano, Italy, 2Laboratory of Medical Genetics, Policlinico S. Orsola-Malpighi, Bologna, Italy, 3Department of MaxilloFacial Surgery San Paolo Universitry Hospital, Milano, Italy, 4Department of

Clinical genetics and Dysmorphology

Maxillo-Facial Surgery San Paolo University Hospital, Milano, Italy.

patients with mild MR, hyperactivity and, occasionally, late onset seizures. ASPM is the most commonly involved gene, explaining roughly 25 to 50% of all cases (MCPH5). Careful clinical, neurological and neuroradiological investigation of 12 patients with homozygous or compound heterozygous mutations in ASPM revealed that 1) phenotype may encompass pyramidal signs and short stature (< -2 SD) ; 2) intelligence may be within normal values (IQ > 70) ; 3) more importantly, simplified gyration pattern with anteroposterior gradient is present in most cases,. Migration defects (heterotopias), and infratentorial anomalies are present in some patients. We investigated regional cerebral organization through 3D morphometric analysis of MRI scans in 7/12 patients and showed that 1) reduction of brain volume affects equally white and grey matter. 2) Reduction of cortical volume (and surface) affects more the caudal part of frontal lobes and the rostral part of parietal areas, whereas 3) mediotemporal areas and basal structures are preserved, which is in accordance with preserved mnseic performances. These data allow us to allow us to broaden the phenotype of MCPH and to redefine its diagnostic criteria. P02.023 Mutations of codon 2085 in the helicase domain of ATRX are recurrent and cause a moderate form of AtRX syndrome

Congenital arhinia is an extremely rare malformation derived from an anomalous embryological development of the nose which normally develops between the third and the tenth week of intra-uterine life. The pathogenesis of this anomaly is not clearly understood. Congenital absence of the nose is often associated with other defects, including ear, palatal and ocular malformations, central nervous system and skeletal anomalies. We report the case of two siblings, a 9-year-old girl and a 2-year-old boy, with congenital total arhinia. Both had a complete absence of external nose and nasal cavities associated with dysmorphic facial features including downslanting of palpebral fissures, maxillary hypoplasia and external ear anomalies. Since congenital absence of the nose has been described in a few patients affected by TreacherCollins Syndrome, we conducted a molecular study of TCOF1 gene on the mother and first child DNA, which were negative for both point mutations and large deletions (PCR and DNA analysis through sequencing of the exons 1-26 and the contiguous intronic sequences). As congenital absence of the nose is an extraordinary occurrence in a family and most cases are sporadic, the recurrence of arhinia in the same family, as described here, suggests a common genetic and hereditary origin. The consanguinity of parents supports the hypothesis of an autosomal recessive disorder; nevertheless, we can not exclude a gonadic mosaicism or an autosomal dominant inheritance, as we are not able to verify if the father had pathological features. P02.021 complex genotypes in arrhythmogenic right ventricular cardiomyopathy patients from North West spain
M. Hermida-Prieto, L. Nez, M. I. Rodrguez-Garca, M. Ortiz, R. BarrialesVilla, E. Maneiro, X. Fernndez, L. Cazn, D. Garca, E. Veira, A. CastroBeiras, L. Monserrat; Instituto Universitario de Ciencias de la Salud-CHUAC, A Corua, Spain.

C. Lacoste1, B. Leheup2, I. Agouti1, D. Mowat3, N. Levy1, C. Badens1; 1 Department of Genetics, Hpital denfants de la Timone, Marseille, France, 2 Department of genetics, Hpital Brabois, Vandoeuvre les Nancy, France, 3 Department of Medical Genetics, Sydney Childrens Hospital, Randswick, Sydney, Australia.

Purpose: Desmosomal genes mutations are detected in nearly 50% of arrhythmogenic right ventricular cardiomyopathy (ARVC) patients from different series. Our objective was to analyze the prevalence of mutations in these genes in an ARVC cohort from North West Spain. Methods: Twenty no related index cases with ARVC diagnosis were clinical evaluated in a dedicated cardiomyopathies clinic. Sequence analysis of main desmosomal genes (PKP2, DSP, DSG2 and DSC2) was performed from fresh blood samples, taken after written consent. Results: We found 10 different mutations in 13 index patients (65%). Four patients (20%) had complex genotypes: 1 was homozygous for a DSC2 gene mutation, 1 was compound heterozygous (two mutations in PKP2 gene) and 2 were double heterozygous (two mutations in different genes). All mutations were novel and two (R375X, S329RfsX351) were present in more than one patient. Both mutations are predicted to produce a truncated peptide. R375X in DSC2 gene was detected in two patients: one was homozygous and developed early and severe phenotype and the other patient had a second mutation in PKP2 gene (S329RfsX351). Moreover, this PKP2 mutation was found in 6 different cases. Conclusions: Nearly, 20% of patients could have complex genotypes in the main desmosomal genes. The complex genotypes could develop early and severe phenotype. North West Spain have a highly prevalence (65%) of ARVC patients with desmosomal gene mutations. P02.022 Further delineation of the cerebral morphology in AsPm-related primitive microcephaly : microcephalia ver ais not a small, normally organized cortex !
S. Passemard1, M. Schaer2, M. El Maleh3, O. Boespflug-Tanguy4, T. Billette de Villemeur5, B. Isidor6, B. Grard1, S. Eliez7, P. Gressens8, A. Verloes1; 1 Dept of Genetics, Robert DEBRE Univ. Hospital, PARIS, France, 2Service Mdico-Pdagogique, Dpartement de psychiatrie, Universit de Genve, Geneva, Switzerland, 3Dept of Medical Imaging, Robert DEBRE Univ. Hospital, PARIS, France, 4Dept of Pediatric Neurology, Robert DEBRE Univ. Hospital, PARIS, France, 5Dept of Pediatric Neurology, Armand TROUSSEAU Univ. Hospital, PARIS, France, 6Dept of Genetics,Nantes Univ. Hospital, Nantes, France, 7Service Mdico-Pdagogique, Dpartement de psychiatrie, Universit de Genve, Genve, Switzerland, 8INSERM U676, Robert DEBRE Univ. Hospital, PARIS, France.

ATRX syndrome is characterised by the association of profound mental retardation, facial dysmorphism, urogenital abnormalities and alpha-thalassemia. A large number of ATRX mutations have been described as causative in ATRX syndrome, the majority being missense mutations. Most of the 80 missense mutations reported so far, have been described in one or two families with exception of the R246C which was found in 20% of the ATRX syndromes. Here, we described mutations localised on codon 2085, in one of the 2 major functional domains, the helicase domain, and associated with a mild form of the disease. Among the patients addressed to our lab for ATRX analysis, we found 5 patients from 3 families carrying a mutation on codon 2085, a highly conserved residue of the helicase domain. The substitution was R2085H in 2 families and R2085C in the last one. Clinical data were as follow: the 5 patients have discrete but characteristic facial dysmorphism and present with mental retardation: all of them acquired autonomous walking between 18 and 24 months, three has no speech skill, one can say words, the last one sentences. None of them have genital abnormalities nor seizure. The 3 mothers are carriers and have highly skewed chromosome X inactivation. No mutation of the codon 2085 was found after testing 250 control chromosomes. In conclusion, missense mutations localised on residue 2085 should clearly be considered as causative mutations even though they lead to moderate and incomplete forms of ATRX syndrome. P02.024 Detection of genomic imbalances by arraycGH in two children with syndromic autism.
D. Avdjieva-Tzavella1, S. Hadjidekova2, B. Rukova2, D. Nesheva2, E. Simeonov3, R. Tincheva1, D. Toncheva2; 1 Department of Clinical Genetics, University Pediatrics Hospital, Medical University, Sofia, Bulgaria, Sofia, Bulgaria, 2Department of Medical Genetics, Medical Faculty, Medical University, Sofia, Bulgaria, 3Clinic of Pediatrics, University Hospital Alexandrovska, Sofia, Bulgaria.

Primary microcephalies (MicroCephaly Primary Hereditary, MCPH) have often been presented as a developmental disorders resulting in homogeneous reduction of grey matter in the cortex, without obvious developmental defect except neuronal apauverishment, occurring in

Autism is a complex behaviorally-defined disorder of the immature brain. It is not a disease but a syndrome with multiple non genetic and/or genetic causes. Autism spectrum disorders (ASD) are conditions which can be either isolated or syndromic, that is associated with other clinical features such as facial dysmorphism, limb or visceral malformations, and growth abnormalities. Currently, diagnosable medical conditions, cytogenetic abnormalities, environmental factors, and single-gene defects associated with autism, together account for 10-20% of cases. We report two children with autistic behavior, mental retardation and dysmorphic features. We have used genomic array CytoChip (BlueGnome, Cambridge, UK), covering the entire genome at a median 565 kb, a resolution optimised to detect pathogenic imbalances while mini-

Clinical genetics and Dysmorphology

mizing polymorphisms. Array CGH- analysis revealed cryptic amplification of 16p11.2 region spanning 1,362 Mb in first patient and an amplification spanning 1,274Mb of 7p12.3 region in second patient. The microdeletion and a reciprocal microduplication of 16p11.2 region carry substantial susceptibility to autism and appear to account for approximately 1% of cases. 7p12.3 is a region of shared genetic association between ASD and attention deficit hyperactivity disorder (ADHD). These data strongly support the idea that only a whole-genome highresolution analysis such as array-CGH is able to provide an accurate diagnosis for chromosomal imbalance in patients with ASD. Confirmatory FISH studies with BAC clones are planned for accurate confirmation of CytoChip result. P02.025** Autism spectrum Disorders: emerging data from copy Number Variations analysis

still undergoing periton dialysis at the end of the evaluation period. Because of the morbidity and mortality associated with renal abnormalities in BBS, this study demonstrates the importance of long-term follow up so that proper care and treatment can be provided. P02.027 A micro-duplication of the centromeric domain of the 11p15.5 imprinted gene cluster is associated with loss of DNA methylation and familial BWs

N. Chiesa1, A. De Crescenzo2, A. Mussa1, G. Baldassarre1, L. Perone3, M. Carella4, M. Cirillo Silengo1, A. Riccio2, G. B. Ferrero1; 1 Department of Paediatrics, University of Torino, Torino, Italy, 2Department of Environmental Science, University of Naples, Caserta, Italy, 3Telethon Institute of Genetics and Medicine,, Naples, Italy, 4IRCCS, Casa Sollievo della Sofferenza, Genetica Medica, San Giovanni Rotondo - Foggia, Italy.

M. Mucciolo1, R. Canitano2, M. Mencarelli1, F. Papa1, E. Katzaki1, A. Marozza1, L. Radice2, C. Castagnini1, L. Dosa1, M. Pollazzon1, G. Hayek2, A. Renieri1, F. Mari1; 1 Medical Genetics-Molecular Biology Department-University of Siena, 53100 Siena, Italy, 2Child Neuropsichiatry, University Hospital of Siena, 53100 Siena, Italy.

Autism Spectrum Disorders (ASDs) have a complex and heterogeneous aetiology with a strong evidence of a genetic involvement. To assess the frequency and type of copy number variations (CNVs) in ASD, a cohort of 95 patients has been selected and analyzed by oligo array-CGH with a functional resolution of nearly 100 kb. Array-CGH resulted negative in 57 patients while in 38 at least one rearrangement was identified. A total of 51 rearrangements was identified: 23 deletions and 28 duplications. Among the 38 patients with CNVs the M: F ratio is higher than expected resulting in 6.6:1. Seven CNVs turned to be pathogenic: one de novo (del Xq12) and 6 located in known autism susceptibility regions (dup15q13.3, dup16p13.1, delXp22.31, del15q11.2, del11p12, dup17q12). The 7 patients with pathogenic CNVs were all males with intellectual disability (ID). The majority presented with congenital anomalies (MCA) and dysmorphisms (57.1%); none suffered from epilepsy. Among the cohort of 57 patients without CNVs the M:F ratio resulted in 4.7:1, as currently reported in ASDs. ID was present in 96.5%; MCA and dysmorphisms were present respectively in 21% and 63.2%. Epilepsy rate was 19.3%. The detection rate of CNVs in our series of patients is 7.4%. Patients with pathogenic CNVs differed from the cohort without any CNVs for the presence of congenital anomalies, more frequent in the first group. Furthermore, the rate of epilepsy found in the all group was 19.3%, while in the subgroup of patients with pathogenic CNVs (n=7) epilepsy was not detected. P02.026 Evaluation of clinical findings in 20 patients with Bardet-Biedl syndrome

Beckwith-Wiedemann syndrome (BWS) is an overgrowth disorder caused by a variety of molecular lesions affecting a two-domain cluster of imprinted genes at chromosome 11p15.5. Most cases are sporadic and result from DNA methylation defects at either one of the two imprinting centres (IC1 and IC2). Here, we describe a familial case with a 166 kb duplication of the centromeric domain and loss of IC2 methylation. The proband is an infant with macroglossia, macrosomia, onfalocele and hexadactylism, her mother and uncle presented with abdominal wall defects. The duplication that comprises IC2 and the 5 part of the anti-sense KCNQ1OT1 gene was identified by MSMLPA (methylation-sensitive multiple-ligation-probe-amplification), confirmed by SNPa analysis and demonstrated to be in-cis by FISH. This genomic lesion cosegregates with BWS phenotype and IC2 hypomethylation, but does not affect IC1 methylation. cDNA sequencing of KCNQ1OT1 demonstrated that both the maternal and paternal alleles were expressed in the proband and her mother, indicating loss of KCNQ1OT1 imprinting. The results obtained are consistent with the presence of a maternal IC2 duplication that results in partial hypomethylation at IC2 and activation of the maternal KCNQ1OT1, likely silencing the imprinted genes of the IC2 domain (e.g. CDKN1C). This is the first familial case of BWS with IC2 hypomethylation in which the molecular defect is identified. Maternal 11p15.5 duplications are generally associated with the Silver-Russell syndrome phenotype and include the CDKN1C gene. On the contrary, in the reported case, the duplication does not include any full-length genes, affects IC2 methylation and causes BWS. P02.028 Microdeletions within the Imprinting region 2 on 11p15 identify a distinct category of Beckwith-Wiedemann syndrome phenotype at risk for cardiac arrythmia.

M. Zollino1, F. Gurrieri1, G. Marangi1, D. Orteschi1, R. Pietrobono1, F. Bellocci2, G. Neri1; 1 Istituto di Genetica Medica, Universit Cattolica del Sacro Cuore, Roma, Italy, 2 Istituto di Cardiologia, Universit Cattolica del Sacro Cuore, Roma, Italy.

O. Altiok Clark1, E. Mihci1,2, S. Akman3; 1 Department of Medical Genetics, Akdeniz University School of Medicine, Antalya, Turkey, 2Department of Pediatrics, Division of Clinical Genetics, Akdeniz University School of Medicine, Antalya, Turkey, 3Department of Pediatric Nephrology, Akdeniz University School of Medicine, Antalya, Turkey.

Bardet-Biedl syndrome (BBS) is a genetically heterogeneous disorder characterized by one or more of the following six major features: retinal degeneration, early-onset obesity, cognitive impairment, genitourinary tract malformations, renal dysfunction and polydactyly. According to recent studies, defects in the basal bodies of ciliated cells appear to be the underlying cause of the syndrome. Although BBS was originally thought to be a recessive disorder, it has since been shown that some forms of BBS are passed on via triallelic inheritance. We retrospectively evaluated 20 patients with BBS because of associated abnormalities. between January 2001 and January 2009. We found both classical and rare findings of BBS in our patients. We identifed a patient with persistent urogenital sinus, urethrovaginal fistule together with BBS as rare findings. Our special attention was given to monitoring those patients with renal abnormalities, because renal disease is a major cause of premature mortality in patients with BBS. We found renal abnormalities in 10 of the patients. The renal abnormalities observed ranged from minimal pelvicalyceal dilatation (PKD) to endstage renal failure. Among of these, four had a polycystic kidney and six had PKD. Due to end-stage renal failure, three of these patients received a renal transplant from a relative. Four of the patients were

Beckwith-Wiedemann syndrome (BWS) is an overgrowth condition characterized by clinical variability and genetic heterogeneity. Psychomotor retardation is uncommon and familiar recurrence rare. BWS is caused by disregulation of two imprinting domains (ICR1 and ICR2) on 11p15, ICR2 being involved in most cases. Abnormalities of ICR2 include loss of methylation on the maternal allele, or point mutations of CDKN1C. We observed a 11p15 microdeletion involving ICR2 in two unrelated females , aged 16 and 19 years, respectively. The deletion spanned about 900 kb in the first case, including CDKN1C, KCNQ1 and additional 15 flanking genes. The size of the deletion in the second patient was 200 kb, encompassing the entire KCNQ1OT1 antisense transcript, ICR2 and part of KCNQ1. Loss-of-function mutations of KCNQ1 can be responsible for long QT syndrome type 1. Both patients presented with an atypical BWS phenotype, including mental retardation in the first patient, a long QT syndrome in the second and overgrowth and facial dysmorphisms in both. The long QT syndrome was responsible for a cluster of life-threatening ventricular fibrillation episodes, needing an urgent implant of intra-cardiac defibrillator. Literature deals with only one additional BWS patient with microdeletion of ICR2. However this event might be under-estimated. A search for a ICR2 microdeletion should be performed in each BWS patient with impaired methylation of ICR2, and patients hemizygous for KCNQ1 should be monitored for long QT syndrome. If a ICR2 microdeletion

Clinical genetics and Dysmorphology

is diagnosed in female patients, a 50% recurrence risk of BWS is expected in the offspring. P02.029 Orofaciodigital syndromes: a case series.

P02.031** Larger cohort of Branchio-oculo-facial syndrome (BOFs) patients: phenotype description and TFAP2A genotype findings

I. Panigrahi, R. R. Das, R. K. Marwaha; Post Graduate Institute of Medical Rducation and Research, Chandigarh, India.

Introduction: Orofaciodigital syndrome (OFD), an inherited syndrome, with varying combinations of defects of the oral cavity, face, and hands, including lobulated or bifid tongue, cleft or pseudocleft palate, tongue tumours, missing or malpositioned teeth, hypoplastic nasal alar cartilage, depressed nasal bridge, bifid nasal tip, brachydactyly, clinodactyly, incomplete syndactyly, and, frequently, mental retardation. They have variable presentations as well as inheritance patterns. Objectives: To study clinical variability of orofaciodigital syndrome in subjects who were seen in genetic clinic in last 3 years. Methods: Subjects with OFDs seen in Genetic unit in APC were subjects of this retro-spective analysis. Results: There were 4 cases with OFD with M:F ratio = 1:3 and the consultations were from age of 4 years to 21 years. On database search, the clinical diagnoses made were: OFD I, OFD II and OFD IX in 3 cases each with 4th subject having overlapping features of both OFD II & VI. The last case had lingual nodules, central polydactyly, bifid thumb, cardiac defect and bilateral renal parenchymal disease. Predominant clinical presentations were developmental delay, abnormal facies, digital anomaly and short stature. Conclusions: Subjects with OFDs can have varying clinical presentation and should be differentiated from craniosynostosis syndromes and oral-cardio-digital syndromes. Digital anomalies may not be present in all. A high clinical suspicion, examination of parents and appropriate investigations are required to arrive at early diagnosis and provide proper genetic counseling. OFD II and OFD VI may represent a clinical continuum of same underlying gene defect. P02.030 two siblings with blepharophimosis, psychomotor delay and severe growth failure: a new autosomal recessive blepharophimosis-mental retardation syndrome?

Y. J. Sznajer1,2, L. Burglen3, S. Blesson4, E. Bieth5, S. Lyonnet6, C. Baumann7, D. Lacombe8, A. David9, E. Galliani10, W. Just11; 1 Hopital Universitaire des Enfants Reine Fabiola, Brussels, Belgium, 2Center for Human Genetics, Brussels, Belgium, 3Service de gntique, Hpital Trousseau, Paris, France, 4Service de gntique, Ple Biologie, CHRU de Tours - Hpital Bretonneau, Tours, France, 5Service de Gntique mdicale; Hpital Purpan, Toulouse, France, 6Dept of Genetics, University Paris Descartes and Hpital Necker Enfants Malades, AP-HP, Paris, France, 7Dept of Clinical Genetics, Hpital Robert Debr, Paris, France, 8Service de Gntique Mdicale, CHU Pellegrin, Bordeaux, France, 9Service de Gntique mdicale, CHU Nantes, Nantes, France, 10Maxillo-Facial/Plastic Surgery Dept, Centre de Rfrence Malformations de la Face, Hpital denfants Armand-Trousseau (AP-HP), Paris, France, 11Human Genetics, University of Ulm, Ulm, Germany.

M. Dentici1, R. Mingarelli1, B. Dallapiccola2; 1 IRCCS-Casa Sollievo della Sofferenza, San Giovanni Rotondo and CSSMendel Institute, Rome, Italy, 2Bambino Ges Pediatric Hospital, IRCCS, Rome, Italy.

Blepharophimosis is characterized by a fixed reduction in the vertical distance between the upper and lower eyelids with short palpebral fissures. It is commonly associated to other periocular anomalies that may occur either as isolated features or as part of multiple congenital anomaly (MCA) syndromes. Many syndromic blepharophimosis patients show variable degree of mental retardation and are referred to as BMR (blepharophimosis mental retardation), a term which includes a clinical and etiological heterogeneous group of disorders. We report on two siblings, a 6 year old girl and a 18 month old male, presenting overlapping clinical findings. Striking facial dysmorphisms included upward slanted palpebral fissures, blepharophimosis, telecanthus, hypertelorism, posteriorly rotated ears with over-folded helices and micrognathia. Ectodermal abnormalities included fine hair, sparse eyebrows and thin skin. Both patients had feeding difficulties with gastroesophageal reflux and growth retardation. Psychomotor skills were severely delayed and with no verbal capacity. The male sib displayed low GH levels, while the older sister had low cholesterol and mildly raised TSH levels. Numerous metabolic/genetic investigations, including cholesterol precursors dosage and high resolution array-CGH, were negative. The clinical history of the siblings was uploaded to the Dysmorphology Diagnostic System (DDS) developed by DYSCERNE. Differential diagnosis with BMR syndromes, including Dubowitz syndrome, Marden-Walker syndrome, Ohdo/Ohdo-like syndromes and cholesterol storage disorders was discussed with the DDS experts which concluded that these two siblings likely represent a previously unreported autosomal recessive-BMR syndrome. This study was supported by DYSCERNE, A European Network of Centres of Expertise for Dysmorphology (DG Sanco), Project 2006122.

BOFS patients develop distinctive features: abnormal external ear; retro-auricular overlying skin; clefts or pseudocleft; eye involvement (microphthalmia or coloboma) and premature grey hair. Patients may have mental retardation and all type of deafness. TFAP2A gene deletion or missense mutations were identified in one familial and 5 sporadic patients (1). Stoetzel et al. confirmed these findings in one family and 3 sporadic patients (2). No precise phenotype criteria was validated to diagnose BOFS. A checklist was elaborated based on the published reports available in BOFS patients. Four familial and 9 sporadic patients with BOFS were included in this study. All patients developed any type of clefts (lip, palate and/or pseudo-). Pre auricular pits/fistula and abnormal external ears were noted in familial presentations as for 6 sporadic patients. Nasolacrymal duct stenosis and malformed nose was diagnosed for two sporadic and one familial presentations. Overlying skin was found in one familial and one sporadic patients. Premature grey hair was noted in three familial and in one sporadic patients. Ophthalmic anomalies were diagnosed in the two familial as in 7 sporadic patients. One family had neurosensorial deafness as 4 sporadic patients. Three sporadic patients developed mental retardation. In all affected patients but one, TFAP2A gene sequencing was diagnostic for missense/nonsense mutations or gene deletion. Hotspot mutations in exons 4 and 5 in the gene encoding the transcription factor AP2A were found. References (1) Milunsky et al. Am J Hum Genet 2008;82:1171-1177 (2) Stoetzel C. et al. Am J Med Genet 2009;149A:2141-2146 P02.032 A new case of blepharophimosis syndrome (BPEs) associated with translocation between chromosomes 3q and 8q
E. Braha1, M. Volosciuc2, V. Gorduza1, C. Bujoran2; 1 University of Medicine and Pharmacy, Iasi, Romania, 2Pediatric Hospital Iasi, Iasi, Romania.

We report a case of 2 month old girl with hort stature, dolichocephaly, blepharophimosis, ptosis and epicanthus inversus. The proband was the first child of a healthy, nonconsanguineous couple. Family anamnesis was negative. Evolution of pregnancy was with hydramnios; preterm delivery at 35 months of age. Chromosomal analisys showed an apparently balanced karyotype with translocation between chromosomes 8q and 3q. Genetic testing for parents should be performed in the context of genetic counseling (the karyotype of the parents - in work). Blepharophimosis syndrome (BPES) is a complex eyelid malformation characterized by four major features: blepharophimosis, ptosis, epicanthus inversus, and telecanthus. The diagnosis of BPES is essentially based on clinical findings. It is possible that our patient has a contiguous gene defect including at least one locus from chromosome 3q for a type of blepharophimosis, further suggesting that multiple loci exist on chromosome 3q for eyelid development. Cytogenetic rearrangements are rare, estimated to occur in 2% of individuals with BPES. [Beysen et al 2009] P02.033 interstitial deletion of chromosome 3q24 to 3q25.33 associated with an unusual brachydactly, facial telangectasia and brisk reflexes
C. M. Brewer1, C. John2, F. Katka3, R. Evans3; 1 Clinical Genetics Department, Peninsula Clinical Genetics Service, Exeter, United Kingdom, 2Wessex Regional Genetics Laboratory, Salisbury, United

Clinical genetics and Dysmorphology

Kingdom, 3Derriford Hospital, Plymouth, United Kingdom.

19p13.13p13.12 deletion encompassing the CACNA1A gene. This microdeletion overlapped the deletion recently reported by Auvin (Epilepsia, 50(11):2501-2505, 2009) concerning a patient presenting infantile spasms, neonatal hypotonia, macrocephaly, advance stature and bone age. In the overlapping region of the 2 deletions is located the CACNA1A gene (calcium channel alpha 1A subunit) encoding a sub-unit of voltage-dependant calcium channel. Mutations of this gene are responsible of familial hemiplegic migraine and episodic ataxia type 2. The haploinsufficiency of CACNA1A probably plays an important role in the phenotype of these 2 patients. P02.036** sHOc2 mutations in patients with cardio-faciocutaneous syndrome

We report a girl with developmental delay, neurological, dermatological and immune features, facial dysmorphism and unusual brachydactyly with a 10.7Mb interstitial deletion within 3q24 to 3q25.33. The 4th and 5th metacarpals, and 3rd and 4th metatarsals are short, consistent with the overlapping conditions of brachydactylys type D and E. She has marked joint laxity and brisk reflexes, facial telangectasia and recurrent infections. Growth is normal. The deleted interval contains 55 annotated genes, but none is an obvious candidate for any of the features observed here. Deletions of this region of chromosome 3 are rare, with only one further case reported in the literature. No photos of the other case were published, but the description is of 1st and 2nd toes that were unusually long compared with the other toes. This sounds remarkably similar to the findings in our case. This case is noteworthy because this case represents a new microdeletion syndrome, and we suggest that an key feature of this may be an unusual form of brachydactyly. P02.034 Brachyphalangy, polydactyly and tibial aplasia/ hypoplasia syndrome (Omim 609945) in a girl born to consanguineous parents

E. M. M. Burkitt Wright1,2, J. Shorto3, B. Kerr1,2; 1 Genetic and Developmental Medicine, Manchester Biomedical Research Centre, St Marys Hospital, Manchester, United Kingdom, 2Medical Genetics Research Group, University of Manchester, Manchester, United Kingdom, 3 Manchester Regional Genetics Laboratory, Central Manchester Hospitals Foundation Trust, Manchester, United Kingdom.

Y. Shafeghati1, K. Kahrizi1, H. Najmabadi1, A. W. Kuss2, H. Ropers2, A. Tzschach2; 1 Genetics Research Center, University of Social Welfare and Rehabilitation Sciences, Tehran, Islamic Republic of Iran, 2Max Planck Institute for Molecular Genetics, Berlin, Germany.

Brachyphalangy, polydactyly and tibial aplasia/hypoplasia syndrome (OMIM 609945) is a rare syndrome characterized by severe limb malformations, facial dysmorphic features and additional clinical problems. Only seven patients have been reported to date, and the etiology of this disorder is unknown. Autosomal dominant inheritance with variable expression has been suggested based on the presence of minor features in some parents and the fact that neither parental consanguinity nor pairs of affected siblings were reported. We report on the first patient with this syndrome who was born to consanguineous parents. Neither the mother nor the father, who were first cousins, had clinical features suggestive of a manifestation of brachyphalangy, polydactyly and tibial aplasia/hypoplasia syndrome. The patient had no siblings, and the family history was unremarkable. Clinical problems included brachydactyly of hands and feet, splaying of fingers and toes, preaxial polydactyly of feet, bilateral tibial aplasia, shortened radius and ulna and characteristic facial dysmorphic signs. Consanguinity in this patient indicates an autosomal recessive etiology, a mode of inheritance that should not be dismissed in genetic counseling as long as the causative gene defect of this disorder remains elusive. P02.035 A 1.6 mb deletion of the 19p13.13p13.12 region detected by array-cGH and encompassing the cAcNA1A gene is responsible of a non evolutive epileptic encephalopathy with macrocephaly, connective tissue dysplasia and urinary reflux.

Cardio-facio-cutaneous syndrome (CFC), Noonan syndrome (NS) and Costello syndrome (CS) show both considerable phenotypic heterogeneity and overlap, and arise due to mutations in several genes whose products interact within the RAS-MAPK pathway. Whilst HRAS mutations cause CS, many different genes (PTPN11, SOS1, KRAS, NRAS, BRAF, MEK1 and MEK2) cause NS and CFC, and a significant proportion of NS/CFC patients have no mutation identified currently. Patients in whom CS is clinically suspected but no HRAS mutation is found are generally considered to have CFC. De novo S2G mutations in SHOC2 were recently reported in patients with a variant NS phenotype, Noonan-like syndrome with loose anagen hair. This mutation generates a myristoylation site, the myristoylated protein then being targeted to the cell membrane, where it interacts with RAS and RAF proteins. Samples from 84 patients referred for molecular diagnostic testing of BRAF, KRAS, MEK1, MEK2 and HRAS because of a clinical suspicion of CFC or CS, in whom no mutation was found in these genes, were screened for the S2G mutation in SHOC2. 11 such mutations were identified, confirming that this mutation is a common and important cause of a CFC/CS/Noonan-like phenotype. Further work to characterise this patient group is underway. These results emphasise the significance of SHOC2 in germline disorders of the RAS-MAPK pathway and indicate that molecular diagnostic testing for the S2G mutation is likely to yield positive results in groups of patients in whom the diagnoses of CFC, CS or severe Noonan syndrome are being considered. P02.037 Desmin mutations as a cause of right ventricular cardiac failure affect the intercalated disk.

F. Amram1, G. Morin1, J. Barois-Guilliot1, R. Gouron1, A. Leke1, B. Demeer1, A. Receveur1, S. Kanafani1, J. Andrieux2, H. Copin1, M. Mathieu1; 1 University Hospital, Amiens, France, 2Jeanne de Flandre Hospital, Lille, France.

E. Otten1, A. Asimaki2, A. Maass1, I. M. van Langen3, A. van der Wal3, N. de Jonge4, M. P. van den Berg1, J. E. Saffitz2, A. A. M. Wilde3,5, J. D. H. Jongbloed1, J. P. van Tintelen1; 1 University Medical Center Groningen, Groningen, Netherlands, 2Beth Israel Deaconess Medical Center, Boston, MA, United States, 3Academic Medical Center, Amsterdam, Netherlands, 4University Medical Center Utrecht, Utrecht, Netherlands, 5Interuniversity Cardiology Institute of the Netherlands, Utrecht, Netherlands.

This female patient is the third daughter of unrelated parents. After a pregnancy characterized by a polyhydramnios during the second trimester and an oligoamnios at the end of the third trimester, the delivery occurred spontaneously at 41 WA. At birth, the patient presented a severe hypotonia, marfanod habitus, hyperelastic skin and severe gastro-oesophageal reflux requiring Nissen surgical treatment. In the first months she presented numerous urinary infections secondary to urinary tract reflux. Dysmorphic features were noticed: high and large forehead, frontal bossing, down slanting palpebral fissures, external strabismus, and arachnodactyly of fingers and toes. The outcome was severe: non evolutive epileptic encephalopathy, absence of speech, absence of walking. She also presented an advance of the stature (+2.5 SD) and the bone age, and a macrocephaly (+2.5 SD). At age 11, a dorsal-lumbar scoliosis was surgically repaired. All etiological investigations were normal: karyotype with FISH 15q11q13, ophtalmological examination, EEG and EMG, cerebral MRI, muscucular and cutaneous biopsy. Array-CGH showed a de novo

Background: Desmin is the main intermediate filament in skeletal and cardiac muscle cells. Abnormal desmin causes skeletal muscle weakness and/or cardiomyopathy. Desmin related myopathy (DRM)(MIM#601419) is associated with dilated (DCM), restrictive or hypertrophic cardiomyopathy. Recently, we observed both DCM and (probable) Arrhythmogenic Right Ventricular Cardiomyopathy (ARVC) in three Dutch families carrying an identical desmin (DES) mutation. This suggests DES mutations to cause cardiomyopathy affecting both the left and right ventricle. Because of this overlap with ARVC, we studied the prevalence of DES mutations in (probable) ARVC patients and we studied the cardiac desmosomal composition in DES mutation carriers. Methods: DES was screened for mutations in 50 ARVC(-like) patients. Besides, immunohistochemistry of desmosomal proteins was performed in myocardial tissue from 3 patients carrying two different DES mutations, causing an ARVC(-like) phenotype in one and a severe biventricular cardiomyopathy in two other patients. Results: No additional DES mutations were found in our patient group.

Clinical genetics and Dysmorphology

Immunohistochemistry demonstrated accumulation of desmin aggregates with abnormally shaped intercalated disks and normal amounts of desmosomal proteins in p.S13F mutation carriers but a decreased amount of connexin 43, desmoplakin and plakophilin 2 in two patients carrying the p.R454W mutation. Conclusions: (Probable) ARVC or severe cardiomyopathy with right ventricular involvement are possible phenotypes in DRM. Moreover, we demonstrated that DES mutations may affect the architecture and/ or the localisation of multiple proteins at the intercalated disk, suggesting a common pathophysiological pathway between arrhythmogenic cardiomyopathies caused by desmosomal gene mutations (mainly affecting the right ventricle) and cardiomyopathies caused by DES mutations. P02.038 The importance of genetic examination in sex assessment in the new born
O. D. Marginean, I. I. Simedrea, A. D. Anghel, E. P. Pavel; 1 Ist Pediatric Clinic, Timisoara, Romania.

7
P02.040 A novel splice mutation in CYP27A1 associated with cerebrotendinous xanthomatosis (and pulverulent cataract)

S. J. Joyce1, R. Bourkiza2, E. Meyer1, A. Reddy1, H. Patel2, M. Chan2, E. R. Maher1; 1 The University of Birmingham, Birmingham, United Kingdom, 2Barts and the London NHS Trust, London, United Kingdom.

Case report: RR, a male infant, 7 months years old, referred in our department, from chirurgery department, for micropenia and facial dimorphism. The child was born at 32 week of gestation, with 1800 g. There was no significant personal or family history for birth defects and genetic disorders. Examinations reveled: weigh 4500gms (< 3th percentile), height of 61 cms (< 3th percentile);dysmorphic facial features (macrocephaly- head circumference of 40 cms, frontal bossing, hypotelorism, prominent eyes, proeminent philtrum), anterior ears, micropenis ( 0.8 cms), small scrotum and no palbably testis, without associated external genitalia ambiguity. According to the clinical appearance small phallus and scrotum the child get a male sex at birth. After inadequate growth of genitalia the parents decided to have a chirurgery procedure for solve the problem. Laboratory and exams reveals : TSH, Ft3, LH, FSH, Testosterone normal, 17OHP (16.9ng/ml), DHEA (1.57ng/ml). Bone age concordant with chronological age. Ultrashall of the pelvis- a uterus may be present; The hyperplasia of the adrenal glands:1/0.9/08 mms. MRI - no evidence of the uterus. Cariotipe 46XX. The DNA testis for Y chromosome - negative for Y chromosome. In this moment refuse to change the social sex of child. Conclusions: 1. The clinical appearance of external genitalia may be not sufficient to give the social sex of children. 2. The micropenis in the infant is a real challenge to diagnose and treat. 3. The congenital adrenal hyperplasia with psudohermafroditism is a rare disease with severe psychological implications for the child and his family. P02.039 cerebrofaciothoracic syndrome: twin presentation
C. Ruggiero, M. Reyes, A. I. Corominas; Centro de Estudios Genticos, Buenos Aires, Argentina.

Cerebrotendinous xanthomatosis (CTX) is a lipid storage metabolism disorder, inherited in an autosomal recessive manner. CTX is characterized by neurological findings (such as cerebellar ataxia and dementia), premature atherosclerosis, tendon xanthomas and cataracts. It is caused by mutations in CYP27A1, leading to sterol 27-hydroxylase deficiency and abnormal cholestanol tissue deposition. A consanguineous Bangladeshi family with three children with pulverulent cataract and global developmental delay were ascertained. Since learning difficulties could also be observed in two of the other siblings who showed no evidence of cataract, the underlying diagnosis and inheritance pattern was unclear. Genetic linkage studies, employing an autozygosity mapping approach, were undertaken to ascertain the underlying genetic cause. The results showed linkage to the chromosomal region 2q35-q36 which contained the CYP27A1 gene. Direct sequencing of CYP27A1 revealed a homozygous splice mutation in all family members with cataracts. This finding confirms the validity of this approach when identifying genes responsible for disorders, even when the aetiology of the condition and diagnosis are uncertain. It highlights the importance of evaluating if CTX is an appropriate diagnosis when the proband has cataracts, especially since the condition can be treated if identified before irreversible brain damage. P02.041 clinical and genetic analysis of the CHD7 gene in Korean patients with cHARGE syndrome

Y. Lee1, S. Kim1, Y. Shin1, J. Kim2, H. Hong1, Y. Lee1, C. Ki2; 1 Soonchunhyang University Bucheon Hospital, Bucheon, Korea, Republic of, 2 Samsung Medical Center, Sungkyunkwan University, Seoul, Korea, Republic of.

Objective: To describe two female dizygotic twins, born to non consanguineous parents by means of assisted fertilization, who had facial dysmorphism, complex anomalies of vertebrae and ribs (such as bifid ribs, hemivertebrae and costal synostosis), and global developmental delay. Methodology: Presentation of two girls who were studied because of facial and thoracic dysmorphism and mental retardation. Conclusion and Discussion: The features are similar to those initially described by Pascual-Castroviejo et al in 1975 and thus represent new cases of cerebrofaciothoracic dysplasia (OMIM 213980). Only 13 cases have been previously described. The information obtained through the few cases described so far suggests that most likely this is a hereditary condition transmitted as an autosomal recessive trait. There is a broad spectrum of differences in the phenotypic presentation between the sisters and also compared with the 13 cases previously described ), which suggest an autosomal recessive form with wide clinical variability. This may reflect an interaction with other loci, or epigenetic modifications. The interaction with a particular intrauterin factor should be discarded in this case, owing to the twinning state of the sibs. We also describe some anomalies unreported to date in this syndrome such as cardiac malformations, affectation of the auditory conduits, umbilical hernia, fifth finger clinodactyly and Arnold Chiari type I.As more patients with this condition are described, the main features of this syndrome are becoming clearer, and also the inheritance pattern.

Background: CHARGE syndrome (CS; OMIM 214800) is a rare autosomal dominant disease associated with coloboma, heart defects, choanal atresia, retarded growth and development, genital abnormalities, and ear anomalies. Mutations in the CHD7 gene have been suggested to be a genetic background of CS. Methods: In this study, sequence analysis of the CHD7 gene was performed in four Korean sporadic CS patients who showed clinical manifestations of typical or atypical CS. Results: Four novel mutations, three nonsense mutations (Ser705X, Arg1069X, and Trp1534X) and one frameshift mutation (Asn603ThrfsX4), were identified in the CHD7 gene in the patients. Congenital heart disease, aplasia of the semicircular canal, sensoryneural hearing loss, and developmental delay were observed in all patients; however, choanal atresia was not seen in any of them. Conclusions: The phenotypic expressions of these Korean patients with CS were different from those of other ethnicities, and allelic heterogeneity was found in the CHD7 gene in Korean patients with CS. Mutation analysis of the CHD7 gene should be performed in Korean subjects to rule out the possibility of CS, even when clinical manifestations are not typical of CS. P02.042** congenital heart defects in patients with a CHD7mutation

N. Janssen1, M. Baardman2, G. J. du Marchie Servaas3, L. Kapusta4, L. H. Hoefsloot5, W. S. Kerstjens-Frederikse1, C. M. A. van Ravenswaaij-Arts1; 1 Dept. Genetics, University Medical Center Groningen, University of Groningen, Groningen, Netherlands, 2Dept. Genetics & EUROCAT, University Medical Center Groningen, University of Groningen, Groningen, Netherlands, 3Dept. Paediatric Cardiology, University Medical Center Groningen, University of Groningen, Groningen, Netherlands, 4Childrens Heart Centre, Radboud University Nijmegen Medical Centre, Nijmegen, Netherlands, 5Dept. Human Genetics, Radboud University Nijmegen Medical Centre, Nijmegen, Netherlands.

Loss of function mutations in the CHD7 gene cause CHARGE-syndrome, an autosomal dominantly inherited combination of multiple congenital malformations, including congenital heart defects (CHDs) in 66-92% of the patients. The pathogenesis and type of CHDs in CHARGE syndrome are still largely unknown. Most malformations in

Clinical genetics and Dysmorphology

CHARGE syndrome are related to the neural crest (NC), which is in line with our observation of a high CHD7 expression in isolated mouse progenitor NC cells. Cardiac NC cells are involved in pharyngeal arch and conotruncal defects. We hypothesise that CHDs in CHARGE syndrome are NC-related. We are collecting detailed clinical information on almost 350 patients with a CHD7-mutation detected by our group, and compared their CHDs with 381 non-syndromic CHD-patients collected between 2004 and 2008 by EUROCAT. The first results show a heart defect in 189 out of 231 CHARGE-patients (82%). Detailed cardiac information is available for 163 of these 189 patients. Of all CHDs 31% were NC-related (e.g. tetralogy of Fallot, right descending aorta), 56% possibly NC-related (e.g. non-specified VSDs) and only 13% were non-NC-related (e.g. muscular and membranous VSDs, hypoplastic left heart). In the control group these figures were 15%, 40% and 45% respectively (p<0.000). These data confirm an important role for the cardiac neural crest in the pathogenesis of CHDs in CHARGE syndrome. P02.043 Dental abnormalities associated with cherubism
L. Preda1, O. Dinca1, A. Bucur1, C. Dragomir2, E. Severin1; 1 Carol Davila Univ Med Pharm, Bucharest, Romania, 2Genetic Lab, Bucharest, Romania.

primary teeth. On panoramic radiographic examination unerupted supernumerary teeth in both maxilla and mandible were seen (total of 19). The probands granmother was also said to have abnormalities in the numbers and eruption of the permanent teeth. In the management of the disease dental procedures, speech therapy, head protection, preventive treatment for osteoporosis are considered. P02.045 Array-cGH detects mosaic tetrasomy 12p (PallisterKillian syndrome) in peripheral blood without invasive skin biopsy
U. Koehler1, M. Locher1, S. Balg1, K. Reiter2, E. Holinski-Feder1; 1 Medizinisch Genetisches Zentrum Mnchen, Mnchen, Germany, 2von Haunersches Kinderspital, Mnchen, Germany.

Cherubism is an abnormal hereditary condition characterized by progressive bilateral swelling of the mandible, especially in children. In some cases of cherubism, the entire jaw swells and the eyes turn up, enhancing the cherubic facial appearance. Frequently, dental abnormalities are associated with cherubism: congenitally missing second and third molars; premature exfoliation of the deciduous teeth and displacement of permanent teeth secondary to the jaw lesions. Mutations in SH3BP2 gene cause cherubism. Case report: A 9-year-old girl, youngest of three siblings, was diagnosed with cherubism based on clinical findings, radiological manifestations and molecular genetic testing. There was no family history of similar findings. Results and conclusions: Clinical examination revealed swellings at angles of mandible, an asymmetrical enlargement of the mandible and chubbiness of the face. Mixed dentition and poor occlusion were noticed. No ocular, respiratory or cardiovascular problems were found. Except her facial appearance, bad alignment of the teeth and maloclusion, the patient did not exhibit any physical abnormality and showed no signs of mental retardation. Panoramic radiograph showed clearly that the facial enlargement is the result of bone changes. Orthopantomogram revealed bilateral multilocular areas of diminished density in the mandible and congenitally missing second and third lower molars. The maxillary dentition was not affected. A mutation was identified in the SH3BP2 gene in proband and her asymptomatic mother. Lack of signs in mother and the difference in clinical presentation between patient and her carrier mother are consistent with incomplete penetrance in females and variable expression of mutation in SH3BP2 gene. P02.044 Familial supernumerary teeth due to cleidocranial dysplasia
D. Belengeanu, D. Stoicanescu, S. Farcas, N. Meszaros; University of Medicine and Pharmacy, Timisoara, Romania.

We report on a 3 month old girl presenting with multiple dysmorphic features including hypotonia, irregular respiration, hypertelorism, long philtrum, prominent forehead, retrognathy, low set, dysplastic ears, and impaired hearing. She is the second child of a non consanguineous couple. Karyotyping of peripheral blood revealed an inconspicuous karyotype. Because of the profound psychomotor retardation at the age of six months, Array-CGH-analysis was requested. Array-CGH with genomic DNA from peripheral blood using 105K oligo arrays, revealed an amplification of the complete short arm of chromosome 12, which led to the assumption of an additional marker chromosome 12 and most probably of a mosaic isochromosome 12p which is common in patients presenting with Pallister-Killian syndrome. Karyotyping of 100 metaphases and interphase FISH analysis on nuclei from cultured T-lymphocytes using a chromosome 12 centromeric probe failed, simply because detection of an isochromosome 12p in stimulated T-lymphocytes is hard to achieve as only 1-2% of lymphocyte metaphases contain the additional isochromosome 12p. The subsequent karyotyping on fibroblasts of a cultured skin biopsy confirmed the presence of an isochromosome 12p (tetrasomy 12p) in 56% of cells. These results refer to Pallister-Killian syndrome and the girl reported shows several correlating features. Up to now, analysis of fibroblasts from invasive skin biopsies was the only opportunity to detect the additional isochromosome 12p in Pallister-Killian syndrome patients. Array-CGH with genomic DNA from peripheral blood, however, is an adequate and powerful tool to investigate children suspected of having Pallister-Killian syndrome. An invasive skin biopsy is no longer required. P02.046 High-Level expression of functional recombinant human coagulation factor Vii in insect cells

N. Masroori1,2, M. Habibi Roudkenar2, R. Halabian2, M. Mohammadipour2, M. Amani2, P. Bahmani2; 1 Islamic Azad University, Science & Research Campus, Tehran, Islamic Republic of Iran, 2Iranian Blood Transfusion Organization, Tehran, Islamic Republic of Iran.

Cleidocranial dysplasia is a rare generalized dysplasia of osseous and dental tissues with an autosomal dominant inheritance. The disorder is characterized by typical facial and dental appearance, skeletal dysplasia and short stature, the two most striking features being hypo or aplastic clavicles and a surprisingly increased number of supernumerary teeth. Intrafamilial variations of the skeletal abnormalities have been described , but those of dental abnormalities are obscure yet. The majority of cases occur due to loss of function mutations in the RUNX2 gene, which encodes for a transcription factor that is essential for osteoblast differentiation and chondrocyte maturation. It is estimated that the proportion of cases caused by a de novo mutation is high. We present the case of a newborn female with skeletal findings consistent with cleidocranial dysplasia, facial dysmorphism with bossing of the forehead, hypertelorism and a depressed nasal bridge, widely open anterior fontanelle, high-arched palate. On lateral and postero-anterior cephalometric radiographs, we could find unclosed coronal suture. Her mother, 31 years old, has short stature, the same dysmorphic features of the face, patent anterior fontanelle, short, broad thumbs, retained

Recombinant coagulation factor VII (FVII) has been introduced as a potential therapeutic intervention in hemophilia patients who produce antibodies against the coagulation factors. Mammalian cell lines provide only low levels of expression, however the Spodoptera frugiperda cell line Sf9 and baculovirus have proved to be a powerful system for high-level expression of recombinant proteins. Due to the lack of endogenous vitamin K-dependent carboxylase, expression of functional FVII through this system is impossible. In the present study, we report a simple but versatile method to overcome the defect for high-level expression of the functional recombinant coagulation FVII in Sf9 cells. This method involves expression of both the human gamma-carboxylase (hGC) and human FVII genes in the host. The cDNAs of hGC and FVII were isolated and cloned into appropriate vectors. Recombinant baculoviruses carrying the human FVII and hGC genes were generated and insect cells were directly coinfected with the recombinant baculoviruses. Expression of the recombinant FVII (rFVII) was verified, purified, and the biological activity was determined. The expression of FVII and hGC mRNAs in coinfected Sf9 cells were detected by RT-PCR and further confirmed by Western blot analysis. A fourfold decrease in clotting time was observed. Whereas no decrease in prothrombin time was noted when the rFVII expressed in the absence of hGC, the results indicate that the expression of hGC confers functional activity of rFVII. Therefore it may possible to express other vitamin Kdependent coagulation factors using this method in the future.

Clinical genetics and Dysmorphology

P02.047 Ophthalmic findings in the Greek isolate of Cohen syndrome. P02.049 P628L and G624D COLA mutations do not cause classic X-linked Alport syndrome but a milder nephropathy resembling thin Basement membrane Nephropathy

S. Douzgou1, J. R. Samples2,3, N. Georgoudi4, M. B. Petersen1; 1 Department of Genetics, Institute of Child Health, Aghia Sophia Childrens Hospital, Athens, Greece, 2Rocky Vista University, Parker, CO, United States, 3 Department of Opthalmology, Oregon Health & Sciences University, Portland, OR, United States, 4Department of Mental Health and Social Welfare, Institute of Child Health, Aghia Sophia Childrens Hospital, Athens, Greece.

Cohen syndrome is a rare condition of mild to moderate developmental delay, characteristic craniofacial features, childhood hypotonia, joint hyperextensibility, neutropenia, and a variety of ophthalmic abnormalities. A high frequency of the syndrome has been observed in a Greek island with 2,000 inhabitants and a high degree of inbreeding. All patients were homozygous for a COH1 deletion of exons 6 to 16, suggesting a founder effect. We present the results of their first systematic ophthalmologic assessment. Myopia and chorioretinal atrophy were present in all patients of this cohort. Yet, in contrast to all groups previously reported, the majority presented corneal changes, independently from age, gender and family history. A pair of sisters, aged 11 and 15 years old, presented bilateral keratoconus. More frequently (86%) than in any other ethnic group, Greek patients presented cataracts that were bilateral and often graded as high as 3, even at a young age. As a whole, the ophthalmic phenotype of the Greek isolate of Cohen syndrome was characterized by the involvement of both the posterior and the anterior eye segment, bilaterally, in the majority of cases (93%). Greek Cohen subjects that share a founder mutation are at a higher risk of developing blindness in respect to those of other ethnicities and genotypes. This study added to the range of visual problems seen in children and adults with Cohen syndrome the finding of thin corneas and highlighted the need for pachymetry measurement as a means of surveillance and prediction of the visual impairment frequently observed. P02.048 Two years experience of VPS1B gene sequencing: confirmation of diagnostic criteria, value of array-CGH in identifying large rearrangements and identification of a new phenotype

P. Demosthenous1, K. Voskarides1, M. Hadjigavriel2, M. Arsali3, C. Patsias3, P. Zirogiannis4, C. Stavrou5, E. Alexopoulos6, A. Pierides7, C. Deltas1; 1 University of Cyprus, Nicosia, Cyprus, 2Department of Nephrology, Larnaca General Hospital, Larnaca, Cyprus, 3Department of Nephrology, Nicosia General Hospital, Nicosia, Cyprus, 4Lamia Hospital, Lamia, Greece, 5 Evangelismos Hospital, Paphos, Cyprus, 6Department of Nephrology, Aristotle University of Thessaloniki, Thessaloniki, Greece, 7Department of Nephrology, Hippocrateon Hospital, Nicosia, Cyprus.

Classical X-linked Alport syndrome (XLAS), caused by mutations in COL4A5, is associated with childhood hematuria progressing to proteinuria, chronic renal failure (CRF) and end-stage kidney disease (ESKD), often accompanied by deafness. Renal biopsies present splitting, lamellation and thickening of the glomerular basement membrane. We studied clinically and genetically two Greek and two Cypriot families with a clinical and pedigree based suspicion of XLAS despite presenting mild characteristics of the syndrome. DNA linkage analysis and direct sequencing resulted in the identification of mutation COL4A5-G624D in the Greek families and COL4A5-P628L mutation in the Cypriot families. The clinical characteristics of most male patients in these four families are surprisingly mild. Four males currying G624D reached ESKD after the age of 39 and one showed Thin Basement Membrane Nephropathy (TBMN, a mild nephropathy caused in many cases by heterozygous COL4A3 or COL4A4 mutations) on Electron Microscope. Another five of nine males with mild mutation P628L developed ESKD between 30 and 57 years while three show mild CRF at similar ages. Biopsy in one of them showed TBMN. Mutations G624D and P628L are very near the 12th natural interruption of the COL4A5 triple helical domain and this may explain the much milder phenotype. Concluding, some COL4A5 mutations may not lead to ESKD until the 40s and 50s. The phenotypic spectrum may extend to milder forms such as TBMN and late-onset ESKD. This wider or milder spectrum of symptoms may be attributed to the position of the mutation relative to the collagenic interruptions. P02.050 Familial congenital diaphragmatic hernia, cystic kidneys and cardiac anomalies: A new X-linked syndrome?

L. Faivre1, P. Sarda2, A. Moncla3, S. El Chedadeh1, P. Callier1, N. Gigot1, D. Lacombe4, L. Burglen5, M. Rio6, P. Edery7, C. Missirian3, C. Popovici3, B. Aral1, C. Thauvin-Robinet1; 1 Gntique, Dijon, France, 2Gntique, Montpellier, France, 3Cytogntique, Marseille, France, 4Gntique, Bordeaux, France, 5Gntique, Trouseau, Paris, France, 6Gntique, Necker, Paris, France, 7Gntique, Lyon, France.

Cohen syndrome is a rare autosomal recessive inherited disorder that results from mutations in the VPS13B gene. Clinical features consist of a combination of mental retardation, facial dysmorphism, post-natal microcephaly, truncal obesity, slender extremities, joint hyperextensibility, myopia, progressive chorioretinal dystrophy and intermittent neutropenia. From the experience of 42 patients (36 families) referred for molecular analysis of VPS13B for suspicion of Cohen syndrome, two mutations were found in 7 families, one mutation was found in 2 families, and none in 27 families. 244K array-CGH revealed two intragenic rearrangements in the two families with only one mutation (one deletion and one duplication). We found that the presence of chorioretinal dystrophy (92% versus 32%, p=0.0023), intermittent neutropenia (92% versus 5%, p<0.001) and postnatal microcephaly (100% versus 48%, p=0.0045) was significantly higher in the group of patients with VPS13B gene mutations compared to the group of patients without mutations. All patients with VPS13B mutations had chorioretinal dystrophy and/or intermittent neutropenia. The Kolehmainen diagnostic criteria provided 100% sensitivity and 77% specificity. Interestingly, we found an adult aged 34 years compound heterozygous for two splicing variants (IVS34+2T_+3AinsT and IVS57+2T>C) with dysmorphic features, normal incisors, microcephaly, severe neutropenia responible of recurrent infections at age 2 years and chorioretinal dystrophy at age 22 years but absence of obesity and mental retardation (total IQ = 100). This observation enlarges the phenotype of the VPS13B gene, but further underscores the importance of the two key features in the indication for a VPS13B gene study.

A. T. Yeung1, S. Keating2, R. Silver3, D. Chitayat1,3; 1 Division of Metabolic and Clinical Genetics, Department of Paediatrics, The Hospital for Sick Children, University of Toronto, Toronto, ON, Canada, 2 Department of Pathology and Laboratory Medicine, Mount Sinai Hospital, University of Toronto, Toronto, ON, Canada, 3The Prenatal Diagnosis and Medical Genetics Program, Department of Obstetrics and Gynecology, Mount Sinai Hospital, University of Toronto, Toronto, ON, Canada.

Congenital diaphragmatic hernia (CDH) occurs in approximately 1 in 3000 live births and is associated with significant morbidity and mortality. Etiologically, CDH is a heterogeneous with most isolated cases being multifactorial. About 50% of the cases are non-isolated, with associated extra-pulmonary anomalies. Some of these cases are single gene disorders and 5-10% have a chromosomal aetiology. We describe two male siblings of non-consanguineous Caucasian parents with CDH, distinctive facial dysmorphism, cardiac anomalies and renal cysts - a constellation of features not consistent with other previously described CDH syndromes. Both siblings exhibited coarse facial features, hypertelorism, brachydactyly, dilatation of the ascending aorta, valvular dysplasia and enlarged cystic kidneys. In addition, sibling two had hepatomegaly, splenomegaly and enlarged thymus as well some intracranial white matter calcification. Karyotype in both siblings was 46, XY and microarray analysis in the second sib was normal. A maternal uncle had died in infancy as the result of CDH. Thus, the pedigree is strongly suggestive of X-linked inheritance. We review the features of other known X-linked CDH syndromes which include Simpson-Golabi-Behmel syndrome, Craniofrontonasal syndrome, Thoracolumbar syndrome and X-linked Cornelia De Lange syndrome. None of these are reconciled by the phenotype in our two cases and indeed, genetic testing for GPC3 to rule out Simpson-Golabi-Behmel syndrome was negative. We believe that this family exhibits a new X-linked genetic syndrome of which congenital diaphragmatic hernia is a principal feature.

Clinical genetics and Dysmorphology

P02.051 molecular karyotyping provides etiological diagnosis in two malformative patients with blurred chromosomal aberrations P02.053 Further case of 12q duplication: Wolf- Hirschhorn syndrome phenocopy or distinct syndrome?

S. P. Hadjidekova1, D. M. Avdjieva-Tzavella2, I. I. Dimova1, B. B. Rukova1, D. V. Nesheva1, R. S. Tincheva2, D. I. Toncheva1; 1 Department of Medical Genetics, Medical University-Sofia, Sofia, Bulgaria, 2 State University Pediatrics Hospital Queen Evdokia, Medical Faculty, Medical University, Sofia, Bulgaria.

Array CGH has proved useful as a diagnostic tool for patients with developmental delay and congenital malformations. We screened 2 patients with ambiguous cytogenetic results in order to disclosure the molecular basis of the mutations. We have used genomic array CytoChip (BlueGnome, Cambridge, UK), covering the entire genome at a median 565Kb. It investigate sub-telomeres at a median 250Kb resolution, reliably detect mosaicism and examine 90 known genetic conditions at a median 100Kb resolution. Case 1. 10 years old girl with Kabuki make-up syndrome characterized by craniofacial dysmorphism: hypertelorism, up slanting palpebral fissures, ptosis, short philtrum, depressed nasal bridge, microretrognathia, preauricular tag; generalized hypotonia, bilateral simian creases, wide spaced nipples, four toes clinodactyly of the toes. The karyotype was 46,XX,add(4)(q35) and array CGH revealed a duplication of (6)(q25.1;q25.3) spanning 6.377Mb as well as a 4q35.1 duplication of 3.815Mb. Case 2. In the second case with karyotype 46,XX,add(17)(q25) array CGH detected duplication of (17)(q24.2;q25.1) region encompassing 9.58Mb in a 1 year old girl with polydactily of toes and fingers and craniofacial dysmorphism: microcephaly, hypertelorism, low-set ears, high palate. Confirmatory FISH studies with BAC clones are planned for accurate confirmation of CytoChip result. The influence of the known genes in the imbalanced regions and their correlation to the phenotype will be discussed. In conclusion, we demonstrate that molecular karyotyping enables a diagnosis in patients with unclear cytogenetic results. Furthermore, we demonstrate the effect of causal CNVs on the development of dysmorphism. P02.052 Associated malformations in patients with limb reduction deficiencies
C. Stoll, Y. Alembik, B. Dott, M. Roth; genetique medicale, strasbourg, France.

M. Bertoli1,2, V. Alesi2, M. Abate3, C. Palmieri3, G. Barrano2, S. Zampatti1,3, M. Frontali3, A. M. Nardone3; 1 Dipartmento di Biopatologia e Diagnostica per Immagine, Universita di Tor Vergata, Roma, Italia, Rome, Italy, 2Ospedale San Pietro Fatebenefratelli, Roma. Centro Ricerche, Unita di Genetica Medica., Rome, Italy, 3Fondazione Policlinico Tor Vergata Roma, U.O.C. Laboratorio di Genetica Medica, Rome, Italy.

Interstitial duplication of short arm of chromosome 12 is a very rare cytogenetic syndrome previously reported as a phenocopy of WolfHirschhorn Syndrome (DallaPiccola et al.,2008). We present a new case of 12q13 duplication in a 6 years old girl with similar dysmorphic features in early infancy, but with a facial phenotype evolving with age and suggesting a possible different gestalt in older patients. She was born at 31weeks of gestation after uneventful pregnancy. Examination in neonatal period showed measurements under 3rd centile, palatoschisis, lacrimal duct obstruction, malar hypoplasia, shallow orbit with exophtalmo,hypertelorism, down-slanting palpebral fissures, high arched eyebrows,cleft palate, corneal opacity. Early motor development was slightly delayed.At the age of 2 some similarities with Wolf- Hirschhorn Syndrome dysmorphic features were noted. At 6 years she presents with an ataxic but autonomous walk and a border line - mild mental retardation. In addition to previous dysmorphic features, she has a longer face, prominent nasal bridge, and prominent superior-orbit ridge. Hands are small and show a thick, slightly redundant and hypercheratinized skin. CGH Array analysis (CytoChipOligo ISCA 4X44K) with a resolution of 75Kb, revealed a duplication of the long arm of chromosome 12 (dup12q13.13). Compared to the previous case, our patient carries a smaller duplication. Although some features of the present patient at birth and at the age of 2 are reminiscent of the Wolf-Hirschhorn Syndrome, the actual clinical picture appears different. Differences with the previously reported case will be discussed in terms of age and of size of duplication region. P02.054 Prenatal diagnosis of Apert syndrome-case report
V. Varlas, L. Turculet, F. Nedelea, G. Peltecu; Clinical Hospital Filantropia, Bucharest, Romania.

Infants with limb reduction deficiencies (LRD) often have other associated congenital malformations. The purpose of this investigation was to assess the prevalence and the types of associated malformations in a defined population. This study included special strengths: each affected child was examined by a geneticist, all elective terminations were ascertained, and the surveillance for malformations was continued until 1 year of age. The associated malformations in infants with LRD were collected in all livebirths, stillbirths and terminations of pregnancy during 26 years in 347,810 consecutive births in the area covered by our population based registry of congenital malformations. Of the 271 LRD infants born during this period, representing a prevalence of 7.8 per 10,000, 57.9% had associated malformations. There were 17(6.3%) patients with chromosomal abnormalities including 10 trisomies 18, and 62 (22.9%) nonchromosomal recognized dysmorphic conditions. There were no predominant recognised dysmorphic conditions, but VA(C)TER(L) association. However numerous recognised dysmorphic conditions were registered. Seventy eight (28.8 %) of the patients were multiply, non syndromic, non chromosomal malformed infants (MCA). Malformations in the urogenital system (n=50, 14.2%), the cardiovascular system (n=40, 11.4%), the central nervous system (n=27, 7.7%), and the digestive system (n=16, 4.6%) were the most common other malformations. The prevalence of associated malformations, which was more than one in two infants, emphasizes the need for a thorough investigation of infants with LRD. A routine screening for other malformations especially cardiovascular system, urogenital system, central nervous system, and digestive system may be considered in infants and in fetuses with LRD.

Apert syndrome is one of the craniosynostosis syndromes. It has an autosomal dominant inheritance and is characterized by the association of craniostenosys and osseous and membranous syndactyly of the four extremities. The incidence has been estimated at 1/50000 births. The craniosynostosis is bicoronal and is present at birth. Most of the cases are de novo mutations, and could be associated with advanced paternal age. We present a case of pregnant woman who came for second opinion of ultrasound investigation for severe anomalies of extremities, detected at 21 weeks of pregnancy. Those was characterized by complete syndactyly of all extremities,with absence of movements and also associated anamaly of cranian shape, bilateral cleft lip and palat. Those dates rised the suspicion of Apert syndrome. It is worth mentioning advance paternal age of 55 years old. Because of the severity of anomalies detected the family decided to interrupt the pregnancy and clinical examination of the fetus confirm the diagnosis of Apert syndrome. P02.055 First cases of craniosynostosis diagnosed by custom molecular microarray (Array cGc)

P. Tavares1, L. Lameiras1, L. Dias1, L. Nunes2, J. Saraiva3, A. Fortuna4, M. Cunha5, M. H. Carreiro6, M. Rosa7, J. S1,3, S. Almeida1,2, A. Palmeiro1, P. Rendeiro1; 1 CGC Genetics, Porto, Portugal, 2Centro Hospitalar de Lisboa Central, E.P.E., Lisboa, Portugal, 3Hospital Peditrico de Coimbra EPE, Coimbra, Portugal, 4 Instituto Nacional de Sade Dr. Ricardo Jorge, IP, Porto, Portugal, 5Centro Hospitalar Trs-os-Montes Alto Douro EPE, V