Papers by Jean J . Latimer
US Patent 20220056447, 2022
US patent 10684293, 2020
In the detection of a complex multifactorial disease, an MN blood group typing is performed on a ... more In the detection of a complex multifactorial disease, an MN blood group typing is performed on a first blood sample of a subject to be tested for a complex multifactorial disease, and on a second blood sample of a control subject having known phenotypes. Erythrocytes from the samples are isovolumetrically sphered, and M alleles and N alleles expressed on surfaces of erythrocyte cells of the first and second blood samples are labeled by contacting the blood samples with anti-M antibodies and anti-N antibodies associated with a marker. Labeled M alleles and labeled N alleles in the first and second blood samples are detected and quantified using the marker, in order to determine an incidence of somatic mutations in Glycophorin A of the patient. The incidence is compared with data pertaining to patients with a known complex multifactorial disease, to diagnose the patient.
Cancer Epidemiology Biomarkers & Prevention
BMC medical genomics, Jan 30, 2018
Nucleotide Excision Repair (NER) is a major pathway of mammalian DNA repair that is associated wi... more Nucleotide Excision Repair (NER) is a major pathway of mammalian DNA repair that is associated with drug resistance and has not been well characterized in acute lymphoblastic leukemia (ALL). The objective of this study was to explore the role of NER in relapsed ALL patients. We hypothesized that increased expression of NER genes was associated with drug resistance and relapse in ALL. We performed secondary data analysis on two sets of pediatric ALL patients that all ultimately relapsed, and who had matched diagnosis-relapse gene expression microarray data (GSE28460 and GSE18497). GSE28460 included 49 precursor-B-ALL patients, and GSE18497 included 27 precursor-B-ALL and 14 T-ALL patients. Microarray data were processed using the Plier 16 algorithm and the 20 canonical NER genes were extracted. Comparisons were made between time of diagnosis and relapse, and between early and late relapsing subgroups. The Chi-square test was used to evaluate whether NER gene expression was altered at...
Screening mammography has had a positive impact on breast cancer mortality but cannot detect all ... more Screening mammography has had a positive impact on breast cancer mortality but cannot detect all breast tumors. In a small study, we confirmed that low power magnetic resonance imaging (MRI) could identify mammographically undetectable tumors by applying it to a high risk population. Tumors detected by this new technology could have unique etiologies and/or presentations, and may represent an increasing proportion of clinical practice as new screening methods are validated and applied A very important aspect of this etiology is genomic instability, which is associated with the loss of activity of the breast cancer-predisposing genes BRCA1 and BRCA2. In sporadic breast cancer, however, there is evidence for the involvement of a different pathway of DNA repair, nucleotide excision repair (NER), which remediates lesions that cause a distortion of the DNA helix, including DNA cross-links. We describe a breast cancer patient with a mammographically undetectable stage I tumor identified in our MRI screening study. She was originally considered to be at high risk due to the familial occurrence of breast and other types of cancer, and after diagnosis was confirmed as a carrier of a Q1200X mutation in the BRCA1 gene. In vitro analysis of her normal breast tissue showed no differences in growth rate or differentiation potential from disease-free controls. Analysis of cultured blood lymphocyte and breast epithelial cell samples with the unscheduled DNA synthesis assay (UDS) revealed no deficiency in nucleotide excision repair (NER). As new breast cancer screening methods become available and cost effective, patients such as this one will constitute an increasing proportion of the incident population, so it is important to determine whether they differ from current patients in any clinically important ways. Despite her status as a BRCA1 mutation carrier, and her mammographically dense breast tissue, we did not find increased cell proliferation or deficient differentiation potential in her breast epithelial cells, which might have contributed to her cancer susceptibility. Although NER deficiency has been demonstrated repeatedly in blood samples from sporadic breast cancer patients, analysis of blood cultured lymphocytes and breast epithelial cells for this patient proves definitively that heterozygosity for inactivation of BRCA1 does not intrinsically confer this type of genetic instability. These data suggest that the mechanism of genomic instability driving the carcinogenic process may be fundamentally different in hereditary and sporadic breast cancer, resulting in different genotoxic susceptibilities, oncogene mutations, and a different molecular pathogenesis.
Life Sciences, 2021
Aims: Gulf War illness (GWI) is thought to be associated with exposures experienced by soldiers d... more Aims: Gulf War illness (GWI) is thought to be associated with exposures experienced by soldiers deployed in the 1991 Gulf War. A major question is how these exposures continue to influence the health of these individu- als three decades later. One potentially permanent effect of such exposures is the induction of genetic muta- tions. We investigated whether veterans with GWI exhibited persistently elevated levels of somatic mutation. Materials and methods: We applied the blood-based glycophorin A (GPA) somatic mutation assay to a cohort of veterans diagnosed with GWI and a set of both concurrent and historic age-matched controls. This assay quanti- fies red blood cells with a phenotype consistent with loss of one allele at the genetic determinant for the MN blood group, the GPA gene.
Key findings: As a population, those affected with GWI exhibited an uninduced mutation frequency at the GPA locus that was effectively twice that observed in controls, a result that was statistically significant. This result was influenced by an increase in the incidence of individuals with aberrantly high mutation frequencies, seemingly higher than would be expected by dose extrapolation and consistent with the induction of localized genomic instability in the hematopoietic bone marrow stem cells. When these “outliers” were removed from consideration, the remaining affected population retained a significantly higher mean allele loss mutation fre- quency, suggesting that both dose-dependent bone marrow genotoxicity and induction of genomic instability are contributing to the elevation in mutation frequency in these affected veterans.
Significance: This study provides evidence that manifestation of GWI is associated with increased cumulative exposure to agents capable of inducing persistent mutations in bone marrow stem cells. Whether these muta- tions are involved in the clinical aspects of the condition or are simply biomarkers of overall exposure has yet to be determined. The increased incidence of genomic instability suggests that this persistent mutation can have important delayed effects on cellular integrity.
Methods in Molecular Biology, 2020
The unscheduled DNA synthesis (UDS) assay measures the ability of a cell to perform global genomi... more The unscheduled DNA synthesis (UDS) assay measures the ability of a cell to perform global genomic nucleotide excision repair (NER). This chapter provides instructions for the application of this technique by creating 6-4 photoproducts and pyrimidine dimers using UV-C (254 nm) irradiation. This procedure is designed specifically for quantification of the 6-4 photoproducts. Repair is quantified by the amount of radioactive thymidine incorporated during repair synthesis after this insult, and radioactivity is evaluated by grain counting after autoradiography. The results have been used to clinically diagnose human DNA repair deficiency disorders, and provide a basis for investigation of repair deficiency in human tissues or tumors. Genomic sequencing to establish the presence of specific mutations is also used now for clinical diagnosis of DNA repair deficiency syndromes. Few functional assays are available which directly measure the capacity to perform NER on the entire genome. Since live cells are required for this assay, explant culture techniques must be previously established. Host cell reactivation (HCR). As discussed in Chap. 28 is not an equivalent technique, as it measures only transcription-coupled repair (TCR) at active genes, a small subset of total NER. Our laboratory also explored the fluorescent label-based Click-iT assay that uses EdU as the label, rather than 3H thymidine. Despite emerging studies in the literature finding this assay to be useful for other purposes, we found that the EdU-based UDS assay was not consistent or reproducible compared with the 3H thymidine-based assay.
Methods in Molecular Biology, 2020
Host cell reactivation (HCR) is a transfection-based assay in which intact cells repair damage (s... more Host cell reactivation (HCR) is a transfection-based assay in which intact cells repair damage (specifically UV damage) localized to exogenous DNA. This paper provides instructions for the application of the technique featuring 2 ways to perform the assay using expression vectors with luciferase and beta galactosidase, as well as with firefly luciferase and renilla luciferase.
Military Medicine, 2019
Introduction: Veterans of the 1991 Gulf War were potentially exposed to a mixture of stress, chem... more Introduction: Veterans of the 1991 Gulf War were potentially exposed to a mixture of stress, chemicals and radiation that may have contributed to the persistent symptoms of Gulf War Illness (GWI). The genotoxic effects of some of these exposures are mediated by the DNA nucleotide excision repair (NER) pathway. We hypothesized that individuals with relatively low DNA repair capacity would suffer greater damage from cumulative geno-toxic exposures, some of which would persist, causing ongoing problems. Materials and Methods: Blood samples were obtained from symptomatic Gulf War veterans and age-matched controls. The unscheduled DNA synthesis assay, a functional measurement of NER capacity, was performed on cultured lymphocytes, and lymphocyte mRNA was extracted and analyzed by sequencing. Results: Despite our hypothesis that GWI would be associated with DNA repair deficiency, NER capacity in lymphocytes from affected GWI veterans actually exhibited a significantly elevated level of DNA repair (p = 0.016). Both total gene expression and NER gene expression successfully differentiated individuals with GWI from unaffected controls. The observed functional increase in DNA repair capacity was accompanied by an overexpression of genes in the NER pathway, as determined by RNA sequencing analysis. Conclusion: We suggest that the observed elevations in DNA repair capacity and NER gene expression are indicative of a "hormetic," i.e., induced or adaptive protective response to battlefield exposures. Normally such effects are short-term, but in these individuals this response has resulted in a long-term metabolic shift that may also be responsible for the persistent symptoms of GWI.
Frontiers in Oncology, 2019
Melatonin has numerous anti-cancer properties reported to influence cancer initiation, promotion,... more Melatonin has numerous anti-cancer properties reported to influence cancer initiation, promotion, and metastasis. With the need for effective hormone therapies (HT) to treat menopausal symptoms without increasing breast cancer risk, co-administration of nocturnal melatonin with a natural, low-dose HT was evaluated in mice that develop primary and metastatic mammary cancer. Individually, melatonin (MEL) and estradiol-progesterone therapy (EPT) did not significantly affect mammary cancer development through age 14 months, but, when combined, the melatonin-estradiol-progesterone therapy (MEPT) significantly repressed tumor formation. This repression was due to effects on tumor incidence, but not latency. These results demonstrate that melatonin and the HT cooperate to decrease the mammary cancer risk. Melatonin and EPT also cooperate to alter the balance of the progesterone receptor (PR) isoforms by significantly increasing PRA protein expression only in MEPT mammary glands. Melatonin significantly suppressed amphiregulin transcripts in MEL and MEPT mammary glands, suggesting that amphiregulin together with the higher PRA:PRB balance and other factors may contribute to reducing cancer development in MEPT mice. Melatonin supplementation influenced mammary morphology by increasing tertiary branching in the mouse mammary glands and differentiation in human mammary epithelial cell cultures. Uterine weight in the luteal phase was elevated after long-term exposure to EPT, but not to MEPT, indicating that melatonin supplementation may reduce estrogen-induced uterine stimulation. Melatonin supplementation significantly decreased the incidence of grossly-detected lung metastases in MEL mice, suggesting that melatonin delays the formation of metastatic lesions and/or decreases aggressiveness in this model of HER2 + breast cancer. Mammary tumor development was similar in EPT
BMC Medical Genomics, 2018
Background: Nucleotide Excision Repair (NER) is a major pathway of mammalian DNA repair that is a... more Background: Nucleotide Excision Repair (NER) is a major pathway of mammalian DNA repair that is associated with drug resistance and has not been well characterized in acute lymphoblastic leukemia (ALL). The objective of this study was to explore the role of NER in relapsed ALL patients. We hypothesized that increased expression of NER genes was associated with drug resistance and relapse in ALL.
Methods: We performed secondary data analysis on two sets of pediatric ALL patients that all ultimately relapsed, and who had matched diagnosis-relapse gene expression microarray data (GSE28460 and GSE18497). GSE28460 included 49 precursor-B-ALL patients, and GSE18497 included 27 precursor-B-ALL and 14 T-ALL patients. Microarray data were processed using the Plier 16 algorithm and the 20 canonical NER genes were extracted. Comparisons were made between time of diagnosis and relapse, and between early and late relapsing subgroups. The Chi- square test was used to evaluate whether NER gene expression was altered at the level of the entire pathway and individual gene expression was compared using t-tests.
Results: We found that gene expression of the NER pathway was significantly increased upon relapse in patients that took 3 years or greater to relapse (late relapsers, P = .007), whereas no such change was evident in patients that relapsed in less than 3 years (early relapsers, P = .180). Moreover, at diagnosis, the NER gene expression of the early relapsing subpopulation was already significantly elevated over that of the late relapsing group (P < .001). This pattern was validated by an ‘NER score’ established by averaging the relative expression of the 20 canonical NER genes. The NER score at diagnosis was found to be significantly associated with disease-free survival in precursor-B-ALL (P < .001).
Conclusion: Patients are over two times more likely to undergo early relapse if they have a high NER score at diagnosis, hazard ratio 2.008, 95% CI (1.256–3.211). The NER score may provide a underlying mechanism for “time to remission”, a known prognostic factor in ALL, and a rationale for differential treatment.
Keywords: Nucleotide excision repair gene, Acute lymphoblastic leukemia, Childhood ALL, Precursor-B-ALL, Early relapse, Late relapse
Photochemistry and Photobiology, 2015
Nucleotide excision repair (NER) is important as a modulator of disease, especially in
constituti... more Nucleotide excision repair (NER) is important as a modulator of disease, especially in
constitutive deficiencies, such as the cancer predisposition syndrome Xeroderma
pigmentosum. We have found profound variation of NER capacity among normal
individuals, between cell-types and during carcinogenesis. NER is a repair system for
many types of DNA damage, and therefore many types of genotoxic carcinogenic
exposures, including ultraviolet light, products of organic combustion, metals, oxidative
stress, etc. Since NER is intimately related to cellular metabolism, requiring
components of both the DNA replicative and transcription machinery, it has a narrow
range of functional viability. Thus, genes in the NER pathway are expressed at the low
levels manifested by, for example, nuclear transcription factors. Since NER activity and
gene expression vary by cell-type, it is inherently epigenetically regulated. Furthermore,
this epigenetic regulation is disregulated during sporadic breast carcinogenesis. Loss of NER is one basis of genomic instability, a required element in cellular transformation, and one that potentially modulates response to therapy. In this paper, we demonstrate differences in NER capacity in eight adult mouse tissues, and place this result into the context of our previous work on mouse extraembryonic tissues, normal human tissues and sporadic early stage human breast cancer.
EMBO Molecular Medicine, 2013
WNT/b-catenin signalling has been suggested to be active in basal-like breast
cancer. However, in... more WNT/b-catenin signalling has been suggested to be active in basal-like breast
cancer. However, in highly aggressive metastatic triple-negative breast cancers
(TNBC) the role of b-catenin and the underlying mechanism(s) for the aggressiveness
of TNBC remain unknown. We illustrate that WNT10B induces transcriptionally
active b-catenin in human TNBC and predicts survival-outcome of
patients with both TNBC and basal-like tumours. We provide evidence that
transgenic murine Wnt10b-driven tumours are devoid of ERa, PR and HER2
expression and can model human TNBC. Importantly, HMGA2 is specifically
expressed during early stages of embryonic mammogenesis and absent when
WNT10B expression is lost, suggesting a developmentally conserved mode of
action. Mechanistically, ChIP analysis uncovered that WNT10B activates canonical
b-catenin signalling leading to up-regulation of HMGA2. Treatment of
mouse and human triple-negative tumour cells with two Wnt/b-catenin pathway
modulators or siRNA to HMGA2 decreases HMGA2 levels and proliferation. We
demonstrate that WNT10B has epistatic activity on HMGA2, which is necessary
and sufficient for proliferation of TNBC cells. Furthermore, HMGA2 expression
predicts relapse-free-survival and metastasis in TNBC patients.
EMBO, Jun 29, 2013
Wnt/b-catenin signalling has been suggested to be active in basal-like breast
cancer. However, i... more Wnt/b-catenin signalling has been suggested to be active in basal-like breast
cancer. However, in highly aggressive metastatic triple-negative breast cancers
(TNBC) the role of b-catenin and the underlying mechanism(s) for the aggressiveness
of TNBC remain unknown. We illustrate that WNT10B induces transcriptionally
active b-catenin in human TNBC and predicts survival-outcome of
patients with both TNBC and basal-like tumours. We provide evidence that
transgenic murine Wnt10b-driven tumours are devoid of ERa, PR and HER2
expression and can model human TNBC. Importantly, HMGA2 is specifically
expressed during early stages of embryonic mammogenesis and absent when
WNT10B expression is lost, suggesting a developmentally conserved mode of
action. Mechanistically, ChIP analysis uncovered that WNT10B activates canonical
b-catenin signalling leading to up-regulation of HMGA2. Treatment of
mouse and human triple-negative tumour cells with two Wnt/b-catenin pathway
modulators or siRNA to HMGA2 decreases HMGA2 levels and proliferation. We
demonstrate that WNT10B has epistatic activity on HMGA2, which is necessary
and sufficient for proliferation of TNBC cells. Furthermore, HMGA2 expression
predicts relapse-free-survival and metastasis in TNBC patients.
Cancer Immunology, Immunotherapy, 2011
Hydroxysteroid (17b) dehydrogenase type 12 (HSD17B12) is a multifunctional isoenzyme functional i... more Hydroxysteroid (17b) dehydrogenase type 12 (HSD17B12) is a multifunctional isoenzyme functional in the conversion of estrone to estradiol (E2), and elongation
of long-chain fatty acids, in particular the conversion of palmitic to archadonic (AA) acid, the precursor of sterols and the inflammatory mediator, prostaglandin E2. Its overexpression together with that of COX-2 in breast carcinoma is associated with a poor prognosis. We have identified the HSD17B12114–122 peptide (IYDKIKTGL) as a naturally presented HLA-A*0201 (HLA-A2)-restricted CD8? T-cell-defined epitope. The HSD17B12114–122 peptide, however, is poorly immunogenic in its in vitro ability to induce peptide-specific CD8? T cells. Acting as an ‘‘optimized peptide’’, a peptide (TYDKIKTGL), which is identical to the HSD17B12114–122 peptide except for threonine at residue 1, was required for inducing in vitro the expansion of CD8? T-cell effectors cross-reactive against the HSD17B12114–122 peptide. In IFN-c ELISPOT assays, these effector cells recognize HSD17B12114–122 peptidepulsed
target cells, as well as HLA-A2? squamous cell carcinoma of the head and neck (SCCHN) and breast carcinoma cell lines overexpressing HSD17B12 and naturally
presenting the epitope. Whereas growth inhibition of a breast carcinoma cell line induced by HSD17B12 knockdown was only reversed by AA, in a similar manner, the
growth inhibition of the SCCHN PCI-13 cell line by HSD17B12 knockdown was reversed by E2 and AA. Our findings provide the basis for future studies aimed at developing cancer vaccines for targeting HSD17B12, which apparently can be functional in critical metabolic pathways involved in inflammation and cancer.
Proceedings National Academy Sci, 2010
The molecular etiology of breast cancer has proven to be remarkably complex. Most individual onco... more The molecular etiology of breast cancer has proven to be remarkably complex. Most individual oncogenes are disregulated in only approximately 30% of breast tumors, indicating that either very few molecular alterations are common to the majority of breast cancers, or that they have not yet been identified. In striking contrast, we now show that 19 of 19 stage I breast tumors tested with the functional unscheduled DNA synthesis assay exhibited a significant deficiency of DNA nucleotide excision repair (NER) capacity relative to normal epithelial tissue from disease-free controls (n = 23). Loss of DNA repair capacity, including the complex, damage-comprehensive NER pathway, results in genomic instability , a hallmark of carcinogenesis. By microarray analysis, mRNA expression levels for 20 canonical NER genes were reduced in representative tumor samples versus normal. Significant reductions were observed in 19 of these genes analyzed by the more sensitive method of RNase protection. These results were confirmed at the protein level for five NER gene products. Taken together, these data suggest that NER deficiency may play an important role in the eti-ology of sporadic breast cancer, and that early-stage breast cancer may be intrinsically susceptible to genotoxic chemotherapeutic agents, such as cis-platinum, whose damage is remediated by NER. In addition, reduced NER capacity, or reduced expression of NER genes, could provide a basis for the development of biomarkers for the identification of tumorigenic breast epithelium. human primary breast cell explant culture | human breast tumor explant culture | breast epithelial attached epispher | hypermutability
Proceedings of the National Academy of Sciences , 2010
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Papers by Jean J . Latimer
Key findings: As a population, those affected with GWI exhibited an uninduced mutation frequency at the GPA locus that was effectively twice that observed in controls, a result that was statistically significant. This result was influenced by an increase in the incidence of individuals with aberrantly high mutation frequencies, seemingly higher than would be expected by dose extrapolation and consistent with the induction of localized genomic instability in the hematopoietic bone marrow stem cells. When these “outliers” were removed from consideration, the remaining affected population retained a significantly higher mean allele loss mutation fre- quency, suggesting that both dose-dependent bone marrow genotoxicity and induction of genomic instability are contributing to the elevation in mutation frequency in these affected veterans.
Significance: This study provides evidence that manifestation of GWI is associated with increased cumulative exposure to agents capable of inducing persistent mutations in bone marrow stem cells. Whether these muta- tions are involved in the clinical aspects of the condition or are simply biomarkers of overall exposure has yet to be determined. The increased incidence of genomic instability suggests that this persistent mutation can have important delayed effects on cellular integrity.
Methods: We performed secondary data analysis on two sets of pediatric ALL patients that all ultimately relapsed, and who had matched diagnosis-relapse gene expression microarray data (GSE28460 and GSE18497). GSE28460 included 49 precursor-B-ALL patients, and GSE18497 included 27 precursor-B-ALL and 14 T-ALL patients. Microarray data were processed using the Plier 16 algorithm and the 20 canonical NER genes were extracted. Comparisons were made between time of diagnosis and relapse, and between early and late relapsing subgroups. The Chi- square test was used to evaluate whether NER gene expression was altered at the level of the entire pathway and individual gene expression was compared using t-tests.
Results: We found that gene expression of the NER pathway was significantly increased upon relapse in patients that took 3 years or greater to relapse (late relapsers, P = .007), whereas no such change was evident in patients that relapsed in less than 3 years (early relapsers, P = .180). Moreover, at diagnosis, the NER gene expression of the early relapsing subpopulation was already significantly elevated over that of the late relapsing group (P < .001). This pattern was validated by an ‘NER score’ established by averaging the relative expression of the 20 canonical NER genes. The NER score at diagnosis was found to be significantly associated with disease-free survival in precursor-B-ALL (P < .001).
Conclusion: Patients are over two times more likely to undergo early relapse if they have a high NER score at diagnosis, hazard ratio 2.008, 95% CI (1.256–3.211). The NER score may provide a underlying mechanism for “time to remission”, a known prognostic factor in ALL, and a rationale for differential treatment.
Keywords: Nucleotide excision repair gene, Acute lymphoblastic leukemia, Childhood ALL, Precursor-B-ALL, Early relapse, Late relapse
constitutive deficiencies, such as the cancer predisposition syndrome Xeroderma
pigmentosum. We have found profound variation of NER capacity among normal
individuals, between cell-types and during carcinogenesis. NER is a repair system for
many types of DNA damage, and therefore many types of genotoxic carcinogenic
exposures, including ultraviolet light, products of organic combustion, metals, oxidative
stress, etc. Since NER is intimately related to cellular metabolism, requiring
components of both the DNA replicative and transcription machinery, it has a narrow
range of functional viability. Thus, genes in the NER pathway are expressed at the low
levels manifested by, for example, nuclear transcription factors. Since NER activity and
gene expression vary by cell-type, it is inherently epigenetically regulated. Furthermore,
this epigenetic regulation is disregulated during sporadic breast carcinogenesis. Loss of NER is one basis of genomic instability, a required element in cellular transformation, and one that potentially modulates response to therapy. In this paper, we demonstrate differences in NER capacity in eight adult mouse tissues, and place this result into the context of our previous work on mouse extraembryonic tissues, normal human tissues and sporadic early stage human breast cancer.
cancer. However, in highly aggressive metastatic triple-negative breast cancers
(TNBC) the role of b-catenin and the underlying mechanism(s) for the aggressiveness
of TNBC remain unknown. We illustrate that WNT10B induces transcriptionally
active b-catenin in human TNBC and predicts survival-outcome of
patients with both TNBC and basal-like tumours. We provide evidence that
transgenic murine Wnt10b-driven tumours are devoid of ERa, PR and HER2
expression and can model human TNBC. Importantly, HMGA2 is specifically
expressed during early stages of embryonic mammogenesis and absent when
WNT10B expression is lost, suggesting a developmentally conserved mode of
action. Mechanistically, ChIP analysis uncovered that WNT10B activates canonical
b-catenin signalling leading to up-regulation of HMGA2. Treatment of
mouse and human triple-negative tumour cells with two Wnt/b-catenin pathway
modulators or siRNA to HMGA2 decreases HMGA2 levels and proliferation. We
demonstrate that WNT10B has epistatic activity on HMGA2, which is necessary
and sufficient for proliferation of TNBC cells. Furthermore, HMGA2 expression
predicts relapse-free-survival and metastasis in TNBC patients.
cancer. However, in highly aggressive metastatic triple-negative breast cancers
(TNBC) the role of b-catenin and the underlying mechanism(s) for the aggressiveness
of TNBC remain unknown. We illustrate that WNT10B induces transcriptionally
active b-catenin in human TNBC and predicts survival-outcome of
patients with both TNBC and basal-like tumours. We provide evidence that
transgenic murine Wnt10b-driven tumours are devoid of ERa, PR and HER2
expression and can model human TNBC. Importantly, HMGA2 is specifically
expressed during early stages of embryonic mammogenesis and absent when
WNT10B expression is lost, suggesting a developmentally conserved mode of
action. Mechanistically, ChIP analysis uncovered that WNT10B activates canonical
b-catenin signalling leading to up-regulation of HMGA2. Treatment of
mouse and human triple-negative tumour cells with two Wnt/b-catenin pathway
modulators or siRNA to HMGA2 decreases HMGA2 levels and proliferation. We
demonstrate that WNT10B has epistatic activity on HMGA2, which is necessary
and sufficient for proliferation of TNBC cells. Furthermore, HMGA2 expression
predicts relapse-free-survival and metastasis in TNBC patients.
of long-chain fatty acids, in particular the conversion of palmitic to archadonic (AA) acid, the precursor of sterols and the inflammatory mediator, prostaglandin E2. Its overexpression together with that of COX-2 in breast carcinoma is associated with a poor prognosis. We have identified the HSD17B12114–122 peptide (IYDKIKTGL) as a naturally presented HLA-A*0201 (HLA-A2)-restricted CD8? T-cell-defined epitope. The HSD17B12114–122 peptide, however, is poorly immunogenic in its in vitro ability to induce peptide-specific CD8? T cells. Acting as an ‘‘optimized peptide’’, a peptide (TYDKIKTGL), which is identical to the HSD17B12114–122 peptide except for threonine at residue 1, was required for inducing in vitro the expansion of CD8? T-cell effectors cross-reactive against the HSD17B12114–122 peptide. In IFN-c ELISPOT assays, these effector cells recognize HSD17B12114–122 peptidepulsed
target cells, as well as HLA-A2? squamous cell carcinoma of the head and neck (SCCHN) and breast carcinoma cell lines overexpressing HSD17B12 and naturally
presenting the epitope. Whereas growth inhibition of a breast carcinoma cell line induced by HSD17B12 knockdown was only reversed by AA, in a similar manner, the
growth inhibition of the SCCHN PCI-13 cell line by HSD17B12 knockdown was reversed by E2 and AA. Our findings provide the basis for future studies aimed at developing cancer vaccines for targeting HSD17B12, which apparently can be functional in critical metabolic pathways involved in inflammation and cancer.
Key findings: As a population, those affected with GWI exhibited an uninduced mutation frequency at the GPA locus that was effectively twice that observed in controls, a result that was statistically significant. This result was influenced by an increase in the incidence of individuals with aberrantly high mutation frequencies, seemingly higher than would be expected by dose extrapolation and consistent with the induction of localized genomic instability in the hematopoietic bone marrow stem cells. When these “outliers” were removed from consideration, the remaining affected population retained a significantly higher mean allele loss mutation fre- quency, suggesting that both dose-dependent bone marrow genotoxicity and induction of genomic instability are contributing to the elevation in mutation frequency in these affected veterans.
Significance: This study provides evidence that manifestation of GWI is associated with increased cumulative exposure to agents capable of inducing persistent mutations in bone marrow stem cells. Whether these muta- tions are involved in the clinical aspects of the condition or are simply biomarkers of overall exposure has yet to be determined. The increased incidence of genomic instability suggests that this persistent mutation can have important delayed effects on cellular integrity.
Methods: We performed secondary data analysis on two sets of pediatric ALL patients that all ultimately relapsed, and who had matched diagnosis-relapse gene expression microarray data (GSE28460 and GSE18497). GSE28460 included 49 precursor-B-ALL patients, and GSE18497 included 27 precursor-B-ALL and 14 T-ALL patients. Microarray data were processed using the Plier 16 algorithm and the 20 canonical NER genes were extracted. Comparisons were made between time of diagnosis and relapse, and between early and late relapsing subgroups. The Chi- square test was used to evaluate whether NER gene expression was altered at the level of the entire pathway and individual gene expression was compared using t-tests.
Results: We found that gene expression of the NER pathway was significantly increased upon relapse in patients that took 3 years or greater to relapse (late relapsers, P = .007), whereas no such change was evident in patients that relapsed in less than 3 years (early relapsers, P = .180). Moreover, at diagnosis, the NER gene expression of the early relapsing subpopulation was already significantly elevated over that of the late relapsing group (P < .001). This pattern was validated by an ‘NER score’ established by averaging the relative expression of the 20 canonical NER genes. The NER score at diagnosis was found to be significantly associated with disease-free survival in precursor-B-ALL (P < .001).
Conclusion: Patients are over two times more likely to undergo early relapse if they have a high NER score at diagnosis, hazard ratio 2.008, 95% CI (1.256–3.211). The NER score may provide a underlying mechanism for “time to remission”, a known prognostic factor in ALL, and a rationale for differential treatment.
Keywords: Nucleotide excision repair gene, Acute lymphoblastic leukemia, Childhood ALL, Precursor-B-ALL, Early relapse, Late relapse
constitutive deficiencies, such as the cancer predisposition syndrome Xeroderma
pigmentosum. We have found profound variation of NER capacity among normal
individuals, between cell-types and during carcinogenesis. NER is a repair system for
many types of DNA damage, and therefore many types of genotoxic carcinogenic
exposures, including ultraviolet light, products of organic combustion, metals, oxidative
stress, etc. Since NER is intimately related to cellular metabolism, requiring
components of both the DNA replicative and transcription machinery, it has a narrow
range of functional viability. Thus, genes in the NER pathway are expressed at the low
levels manifested by, for example, nuclear transcription factors. Since NER activity and
gene expression vary by cell-type, it is inherently epigenetically regulated. Furthermore,
this epigenetic regulation is disregulated during sporadic breast carcinogenesis. Loss of NER is one basis of genomic instability, a required element in cellular transformation, and one that potentially modulates response to therapy. In this paper, we demonstrate differences in NER capacity in eight adult mouse tissues, and place this result into the context of our previous work on mouse extraembryonic tissues, normal human tissues and sporadic early stage human breast cancer.
cancer. However, in highly aggressive metastatic triple-negative breast cancers
(TNBC) the role of b-catenin and the underlying mechanism(s) for the aggressiveness
of TNBC remain unknown. We illustrate that WNT10B induces transcriptionally
active b-catenin in human TNBC and predicts survival-outcome of
patients with both TNBC and basal-like tumours. We provide evidence that
transgenic murine Wnt10b-driven tumours are devoid of ERa, PR and HER2
expression and can model human TNBC. Importantly, HMGA2 is specifically
expressed during early stages of embryonic mammogenesis and absent when
WNT10B expression is lost, suggesting a developmentally conserved mode of
action. Mechanistically, ChIP analysis uncovered that WNT10B activates canonical
b-catenin signalling leading to up-regulation of HMGA2. Treatment of
mouse and human triple-negative tumour cells with two Wnt/b-catenin pathway
modulators or siRNA to HMGA2 decreases HMGA2 levels and proliferation. We
demonstrate that WNT10B has epistatic activity on HMGA2, which is necessary
and sufficient for proliferation of TNBC cells. Furthermore, HMGA2 expression
predicts relapse-free-survival and metastasis in TNBC patients.
cancer. However, in highly aggressive metastatic triple-negative breast cancers
(TNBC) the role of b-catenin and the underlying mechanism(s) for the aggressiveness
of TNBC remain unknown. We illustrate that WNT10B induces transcriptionally
active b-catenin in human TNBC and predicts survival-outcome of
patients with both TNBC and basal-like tumours. We provide evidence that
transgenic murine Wnt10b-driven tumours are devoid of ERa, PR and HER2
expression and can model human TNBC. Importantly, HMGA2 is specifically
expressed during early stages of embryonic mammogenesis and absent when
WNT10B expression is lost, suggesting a developmentally conserved mode of
action. Mechanistically, ChIP analysis uncovered that WNT10B activates canonical
b-catenin signalling leading to up-regulation of HMGA2. Treatment of
mouse and human triple-negative tumour cells with two Wnt/b-catenin pathway
modulators or siRNA to HMGA2 decreases HMGA2 levels and proliferation. We
demonstrate that WNT10B has epistatic activity on HMGA2, which is necessary
and sufficient for proliferation of TNBC cells. Furthermore, HMGA2 expression
predicts relapse-free-survival and metastasis in TNBC patients.
of long-chain fatty acids, in particular the conversion of palmitic to archadonic (AA) acid, the precursor of sterols and the inflammatory mediator, prostaglandin E2. Its overexpression together with that of COX-2 in breast carcinoma is associated with a poor prognosis. We have identified the HSD17B12114–122 peptide (IYDKIKTGL) as a naturally presented HLA-A*0201 (HLA-A2)-restricted CD8? T-cell-defined epitope. The HSD17B12114–122 peptide, however, is poorly immunogenic in its in vitro ability to induce peptide-specific CD8? T cells. Acting as an ‘‘optimized peptide’’, a peptide (TYDKIKTGL), which is identical to the HSD17B12114–122 peptide except for threonine at residue 1, was required for inducing in vitro the expansion of CD8? T-cell effectors cross-reactive against the HSD17B12114–122 peptide. In IFN-c ELISPOT assays, these effector cells recognize HSD17B12114–122 peptidepulsed
target cells, as well as HLA-A2? squamous cell carcinoma of the head and neck (SCCHN) and breast carcinoma cell lines overexpressing HSD17B12 and naturally
presenting the epitope. Whereas growth inhibition of a breast carcinoma cell line induced by HSD17B12 knockdown was only reversed by AA, in a similar manner, the
growth inhibition of the SCCHN PCI-13 cell line by HSD17B12 knockdown was reversed by E2 and AA. Our findings provide the basis for future studies aimed at developing cancer vaccines for targeting HSD17B12, which apparently can be functional in critical metabolic pathways involved in inflammation and cancer.
X-ray mammography screening is the current mainstay for early breast cancer detection. It has been proven to detect breast cancer at an earlier stage and to reduce the number of women dying from the disease. However, it has a number of limitations.
These current limitations in early breast cancer detection technology are driving a surge of new technological developments, from modifications of x-ray mammography such as computer programs that can indicate suspicious areas, to newer methods of detection such as magnetic resonance imaging (MRI) or biochemical tests on breast fluids. To explore the merits and drawbacks of these new breast cancer detection techniques, the Institute of Medicine of the National Academy of Sciences convened a committee of experts. During its year of operation, the committee examined the peer-reviewed literature, consulted with other experts in the field, and held two public workshops.
In addition to identifying promising new technologies for early detection, the committee explored potential barriers that might prevent the development of new detection methods and their common usage. Such barriers could include lack of funding from agencies that support research and lack of investment in the commercial sector; complicated, inconsistent, or unpredictable federal regulations; inadequate insurance reimbursement; and limited access to or unacceptability of breast cancer detection technology for women and their doctors. Based on the findings of their study, the committee prepared a report entitled Mammography and Beyond: Developing Technology for Early Detection of Breast Cancer, which was published in the spring of 2001. This is a non- technical summary of that report.
This book reviews the dramatic expansion of breast cancer awareness and screening, examining the capabilities and limitations of current and emerging technologies for breast cancer detection and their effectiveness at actually reducing deaths. The committee discusses issues including national policy toward breast cancer detection, roles of public and private agencies, problems in determining the success of a technique, availability of detection methods to specific populations of women, women's experience during the detection process, cost-benefit analyses, and more.Examining current practices and specifying research and other needs, Mammography and Beyond will be an indispensable resource to policy makers, public health officials, medical practitioners, researchers, women's health advocates, and concerned women and their families.
COMMITTEE ON TECHNOLOGIES FOR THE EARLY DETECTION OF BREAST CANCER
JOYCE C. LASHOF, M.D., FACP, CHAIR, Professor Emerita, School of Public Health, University of California at Berkeley, Berkeley, CA
I. CRAIG HENDERSON, M.D., VICE CHAIR, Adjunct Professor of Medicine, University of California at San Francisco, San Francisco, CA
D. CRAIG ALLRED, M.D., Professor of Pathology, Baylor College of Medicine, Houston, TX
WADE M. AUBRY, M.D., Vice President, The Lewin Group, Associate Clinical Professor of Medicine, University of California at San Francisco, San Francisco, CA
JANET K. BAUM, M.D., FACR, Associate Professor of Radiology, Harvard Medical School, Director, Breast Imaging, Beth Israel Deaconess Medical Center, Boston, MA
SUZANNE W. FLETCHER, M.D., M.Sc., Professor of Ambulatory Care and Prevention, Harvard School of Medicine, Harvard Pilgrim Health Care, Professor of Epidemiology, Harvard School of Public Health, Boston, MA
MARTHE R. GOLD, M.D., M.P.H., Chair, Department of Community Health and Social Medicine, City University of New York Medical School, New York, NY
LEON GORDIS, M.D., D.P.H., Professor of Epidemiology, Johns Hopkins School of Public Health & Hygiene, Baltimore, MD
DANIEL F. HAYES, M.D., Clinical Director, Breast Cancer Program, Lombardi Cancer Center, Georgetown University Medical Center, Washington, DC
CAROLINA HINESTROSA, M.A., Cofounder and Executive Director, Nueva Vida, Silver Spring, MD
JEAN J. LATIMER, Ph.D., Investigator, Magee-Womens Research Institute, Assistant Professor, Department of Obstetrics, Gynecology, and Reproductive Sciences, University of Pittsburgh, Pittsburgh, PA
RICHARD R. NELSON, Ph.D., George Blumenthal Professor, School of International and Public Affairs, Columbia University, New York, NY
KENNETH OFFIT, M.D., M.P.H., Chief, Clinical Genetics Service, Department of Human Genetics, Memorial Sloan-Kettering Cancer Center, New York, NY
FAINA SHTERN, M.D., Director, Office of Research Affairs, Department of Radiology, Beth Israel-Deaconess Medical Center, Harvard Medical School, Boston, MA
MICHAEL W. VANNIER, M.D., Professor and Head, Department of Radiology, University of Iowa College of Medicine, Iowa City, Iowa
DEREK VAN AMERONGEN, M.D., M.S., FACOG, Chief Medical Officer, Humana/ Choice Care, Cincinnati, OH
Liaison for the National Cancer Policy Board
ROBERT DAY, M.D., M.P.H., Ph.D., Emeritus President and Director,
Fred Hutchinson Cancer Research Center, Seattle, WA
Consultants
LARRY NORTON, M.D., Chief, Solid Tumors, Memorial Sloan- Kettering, New York, NY
BARRON LERNER, M.D., Ph.D, Assistant Professor of Medicine and Public Health (in the Center for the Study of Society and Medicine), Columbia University, New York, NY
Develop an approach for identifying chemicals that may contribute to the development or progression of breast cancer,
Identify research needs and recommend improvements to existing test methods, and Pilot a model process that can be applied to other disease endpoints, enabling the ultimate aim of producing a comprehensive approach for identifying hazardous chemicals. Drawing on the fields of cancer biology, toxicology, medicine, epidemiology, public health, and public policy (Figure 1), a multidisciplinary expert panel (Panel) reviewed existing methods for chemical toxicity testing and developed a testing scheme, called the Hazard Identification Approach. This approach provides a methodology for the identification of substances that could elevate breast cancer risk.
Committee members:
Ruthann Rudel, MS, Lauren Zeise, PhD
Joseph Guth, PhD, JD. Gina Solomon, MD, MPH Dale Johnson, PharmD, PhD Shanaz Dairkee, PhD, Rachel Morello-Frosch, PhD Suzanne Fenton, PhD, Ron Melnick, PhD
Jean Latimer, PhD, Vincent James Cogliano, PhD
A report of the Breast Cancer and Chemicals Policy Project, produced by the University of California, Berkeley and the Natural Resources Defense Council, with funding from the California Breast Cancer Research Program, University of California Office of the President