Papers by Salvatore Piscuoglio
Abstract S6-06: The genomic landscape of male breast cancers
Cancer Research, 2015
The Journal of Pathology, 2015
Acinic cell carcinoma (ACC) of the breast is a rare form of triple-negative (that is, estrogen re... more Acinic cell carcinoma (ACC) of the breast is a rare form of triple-negative (that is, estrogen receptor-negative, progesterone receptor-negative, HER2-negative) salivary gland-type tumor displaying serous acinar differentiation. Despite its triple-negative phenotype, breast ACCs are reported to have an indolent clinical behavior. Here, we sought to define whether ACCs have a mutational repertoire distinct from that of other triple-negative breast cancers (TNBCs). DNA was extracted from microdissected formalin-fixed paraffin-embedded sections of tumor and normal tissue from two pure and six mixed breast ACCs. Each tumor component of the mixed cases was microdissected separately. Tumor and normal samples were subjected to targeted capture massively parallel sequencing targeting all exons of 254 genes, including genes most frequently mutated in breast cancer and related to DNA repair. Selected somatic mutations were validated by targeted amplicon resequencing and Sanger sequencing. Akin to other forms of TNBC, the most frequently mutated gene found in breast ACCs was TP53 (one pure and six mixed cases). Additional somatic mutations affecting breast cancer-related genes found in ACCs included PIK3CA, MTOR, CTNNB1, BRCA1, ERBB4, ERBB3, INPP4B and FGFR2. Copy number alteration analysis revealed complex patterns of gains and losses similar to those of TNBCs. Of the mixed cases analyzed,
A survey of DICER1 hotspot mutations in ovarian and testicular sex cord-stromal tumors
Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc, Jan 2, 2015
Sertoli-Leydig cell tumors are characterized by the presence of somatic DICER1 hotspot mutations.... more Sertoli-Leydig cell tumors are characterized by the presence of somatic DICER1 hotspot mutations. In this study, we sought to define the association between DICER1 hotspot mutations and different morphologic subtypes of ovarian Sertoli-Leydig cell tumors. Furthermore, we aimed to assess whether DICER1 hotspot mutations occur in other ovarian sex cord-stromal tumors, testicular sex cord-stromal tumors, or other female genital tract tumors with rhabdomyosarcomatous differentiation. We subjected a series of ovarian Sertoli-Leydig cell tumors (n=32), Sertoli cell tumors (n=5) and gynandroblastomas (n=5), testicular sex cord-stromal tumors (n=15) and a diverse group of female genital tract tumors with rhabdomyosarcomatous morphology (n=10) to DICER1 hotspot mutation analysis using Sanger sequencing. We also tested two gynandroblastomas for the presence of FOXL2 hotspot mutations (p.C134W; c.402C>G). Twenty of 32 (63%) Sertoli-Leydig cell tumors harbored a DICER1 hotspot mutation, of w...
Histopathology, 2015
Polymorphous low-grade adenocarcinoma (PLGA) is the second most common intra-oral salivary gland ... more Polymorphous low-grade adenocarcinoma (PLGA) is the second most common intra-oral salivary gland malignancy. The vast majority of PLGAs harbor a PRKD1 E710D hotspot somatic mutation or somatic rearrangements of PRKD1, PRKD2 or PRKD3. Given the kinase domain homology among PRKD1, PRKD2 and PRKD3, we sought to define whether PLGAs lacking PRKD1 somatic mutations or PRKD gene family rearrangements would be driven by somatic mutations affecting the kinase domains of PRKD2 or PRKD3. DNA was extracted from 8 microdissected PLGAs lacking PRKD1 somatic mutations or PRKD gene family rearrangements. Samples were thoroughly centrally reviewed, microdissected and subjected to Sanger sequencing of the kinase domains of PRKD2 and PRKD3 genes. None of the PLGAs lacking PRKD1 somatic mutations or PRKD gene family rearrangements harbored somatic mutations in the kinase domains of PRKD2 or PRKD3 genes. PLGAs lacking PRKD1 somatic mutations or PRKD gene family rearrangements are unlikely to harbor som...
Journal of Clinical Pathology, 2015
Sequence Directed Nature of Human Mutations (Supplementary Material)
Massively Parallel Sequencing Analysis of Acinic Cell Carcinoma of the Breast
Background: Acinic cell carcinoma (AciCC) is a rare salivary gland-type tumor of the breast, disp... more Background: Acinic cell carcinoma (AciCC) is a rare salivary gland-type tumor of the breast, displaying serous acinar differentiation. Despite its triple-negative phenotype, AciCCs are reported to have an indolent clinical behavior. We sought to investigate whether breast AciCCs would be underpinned by genetic alterations found in 254 breast cancer-related genes, and to define whether AciCCs would have a mutational repertoire distinct from that of other triple-negative breast cancers (TNBCs). Design: Three pure and seven mixed breast AciCCs (six AciCC-invasive ductal carcinomas of no special type and one AciCC-metaplastic carcinoma) were included in this study. DNA was extracted from microdissected formalin-fixed paraffin-embedded sections of tumor and normal tissue. In mixed cases each component was microdissected separately. DNA of sufficient quantity was obtained from 2 pure AciCCs, from the AciCC component of 7 mixed cases and from the non-AciCC components of 4 cases. Massively ...
Morphologic and Genetic Heterogeneity in Mixed Endometrial Carcinomas
Background: Mixed endometrial carcinomas (MECs) comprise a heterogeneous group of tumors characte... more Background: Mixed endometrial carcinomas (MECs) comprise a heterogeneous group of tumors characterized by an admixture of two or more distinct histologic subtypes of endometrial carcinoma. The genetic underpinning of MECs has yet to be fully established. We investigated the repertoire of somatic genetic alterations in 4 MECs by targeted capture sequencing of 341 cancer-related genes in each of their distinct histologic components. Design: Six gynecologic pathologists reviewed slides from 13 tumors originally diagnosed as MECs; four tumors were unanimously diagnosed as bona fide MECs. Representative sections from each case were cut and subjected to microdissection, either laser-assisted or with a needle under a stereo-microscope. DNA was extracted from each tumor component and matched normal tissue and subjected to massively parallel sequencing using the MSK-IMPACT platform that targets the coding regions of 341 actionable cancer-related genes. Somatic single nucleotide variants were...
EWS16T Is a Useful Marker for the Detection of Microsatellite Instability in Endometrial Cancer
Massively Parallel Sequencing Reveals That Microglandular Adenosis Is a Clonal Neoplastic Lesion of Triple-Negative Phenotype
Background: Microglandular adenosis (MGA) is a lesion composed of small infiltrating glands lined... more Background: Microglandular adenosis (MGA) is a lesion composed of small infiltrating glands lined by S100-positive, estrogen receptor (ER)-negative epithelial cells and lacking a myoepithelial cell layer. Although classified as a benign epithelial proliferation, there is evidence to suggest that MGA may constitute a non-obligate precursor of triple-negative breast cancer (TNBC). We sought to define the mutational landscape of MGAs and of TNBCs arising in MGA, and to determine whether MGAs may constitute the substrate from which the TNBCs originated. Design: Six cases of MGAs and three of atypical MGAs (AMGAs) associated with in situ or invasive TNBC were collected. DNA from distinct morphologic components and matched normal tissue was extracted from microdissected representative sections and subjected to massively parallel sequencing targeting all coding regions of 273 genes recurrently mutated in breast cancer or related to DNA repair. Single nucleotide variants were detected by Mu...
Genome Biology, 2015
Background: HER2 is overexpressed and amplified in approximately 15% of invasive breast cancers, ... more Background: HER2 is overexpressed and amplified in approximately 15% of invasive breast cancers, and is the molecular target and predictive marker of response to anti-HER2 agents. In a subset of these cases, heterogeneous distribution of HER2 gene amplification can be found, which creates clinically challenging scenarios. Currently, breast cancers with HER2 amplification/overexpression in just over 10% of cancer cells are considered HER2-positive for clinical purposes; however, it is unclear as to whether the HER2-negative components of such tumors would be driven by distinct genetic alterations. Here we sought to characterize the pathologic and genetic features of the HER2-positive and HER2-negative components of breast cancers with heterogeneous HER2 gene amplification and to define the repertoire of potential driver genetic alterations in the HER2-negative components of these cases. Results: We separately analyzed the HER2-negative and HER2-positive components of 12 HER2 heterogeneous breast cancers using gene copy number profiling and massively parallel sequencing, and identified potential driver genetic alterations restricted to the HER2-negative cells in each case. In vitro experiments provided functional evidence to suggest that BRF2 and DSN1 overexpression/amplification, and the HER2 I767M mutation may be alterations that compensate for the lack of HER2 amplification in the HER2-negative components of HER2 heterogeneous breast cancers. Conclusions: Our results indicate that even driver genetic alterations, such as HER2 gene amplification, can be heterogeneously distributed within a cancer, and that the HER2-negative components are likely driven by genetic alterations not present in the HER2-positive components, including BRF2 and DSN1 amplification and HER2 somatic mutations.
Response to dual HER2 blockade in a patient with HER3-mutant metastatic breast cancer
Annals of Oncology, 2015
HER3 activating mutations have been shown in preclinical models to be oncogenic and ligand-indepe... more HER3 activating mutations have been shown in preclinical models to be oncogenic and ligand-independent, but to depend on kinase-active HER2. Whole-exome sequencing of the primary HER2-negative breast cancer and its HER2-negative synchronous liver metastasis from a 46-year-old female revealed the presence of an activating and clonal HER3 G284R mutation. HER2 dual blockade with trastuzumab and lapatinib as third-line therapy led to complete metabolic response in 2 weeks and confirmed radiological partial response after 8 weeks. Following the resection of the liver metastasis, the patient remains disease-free 40 weeks after initiation of the HER2 dual blockade therapy. Immunohistochemical analysis demonstrated a substantial reduction of phospho-rpS6 and phospho-AKT in the post-therapy biopsy of the liver metastasis. This is the first-in-man evidence that anti-HER2 therapies are likely effective in breast cancers harboring HER3 activating mutations.
High-Resolution Breakpoint Analysis Provides Evidence for the Sequence-Directed Nature of Genome Rearrangements in Hereditary Disorders
Human Mutation, 2015
Although most of the pertinent data on the sequence-directed processes leading to genome rearrang... more Although most of the pertinent data on the sequence-directed processes leading to genome rearrangements (GRs) have come from studies on somatic tissues, little is known about GRs in the germ line of patients with hereditary disorders. This study aims at identifying DNA motifs and higher order structures of genome architecture, which can result in losses and gains of genetic material in the germ line. We first identified candidate motifs by studying 112 pathogenic germ-line GRs in hereditary colorectal cancer patients, and subsequently created an algorithm, termed recombination type ratio, which correctly predicts the propensity of rearrangements with respect to homologous versus nonhomologous recombination events.
90 HOXA13 and Hottip Expression Levels Predict Patients' Survival and Metastasis Formation in Hepatocellular Carcinoma
Journal of Hepatology, 2013
International Journal of Cancer, 2011
Hepatology, 2014
Hepatocellular carcinoma (HCC) is among the leading causes of cancer-related death. Despite the a... more Hepatocellular carcinoma (HCC) is among the leading causes of cancer-related death. Despite the advances in diagnosis and management of HCC, the biology of this tumor remains poorly understood. Recent evidence highlighted long noncoding RNAs (lncRNAs) as crucial determinants of HCC development. In this study we report the lncRNA HOXA transcript at the distal tip (HOTTIP) as significantly up-regulated in HCC specimens. The HOTTIP gene is located in physical contiguity with HOXA13 and directly controls the HOXA locus gene expression by way of interaction with the WDR5/MLL complex. HOX genes encode transcription factors regulating embryonic development and cell fate. We previously described HOX genes deregulation to be involved in hepatocarcinogenesis. Indeed, we observed the marked up-regulation of HOXA13 in HCC. Here, by correlating clinicopathological and expression data, we demonstrate that the levels of HOTTIP and HOXA13 are associated with HCC patients' clinical progression and predict disease outcome. In contrast to the majority of similar studies, our data were obtained from snap-frozen needle HCC biopsies (n 5 52) matched with their nonneoplastic counterparts collected from patients who had not yet received any HCC-tailored therapeutic treatments at the time of biopsy. In addition, taking advantage of gain and loss of function experiments in liver cancer-derived cell lines (HuH-6 and HuH-7), we uncover a novel bidirectional regulatory loop between HOTTIP/HOXA13. Conclusion: Our study highlights the key role of HOTTIP and HOXA13 in HCC development by associating their expression with metastasis and survival in HCC patients, provides novel insights on the function of lncRNA-driven hepatocarcinogenesis, and paves the way for further investigation about the possible role of HOTTIP as a predictive biomarker of HCC. (HEPATOLOGY 2014;59:911-923)
Abstract 4496: UBCH10 overexpression in human colorectal cancers
Cancer Research, 2012
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Papers by Salvatore Piscuoglio