Citations

Abstract Background: A recent association study identified a common variant (rs9790517) at 4q24 to be associated with breast cancer risk. Independent association signals and potential functional variants in this locus have not been explored. Methods: We conducted a fine-mapping analysis in 55,540 breast cancer cases and 51,168 controls from the Breast Cancer Association Consortium. Results: Conditional analyses identified two independent association signals among women of European ancestry, represented by rs9790517 [conditional P = 2.51 × 10−4; OR, 1.04; 95% confidence interval (CI), 1.02–1.07] and rs77928427 (P = 1.86 × 10−4; OR, 1.04; 95% CI, 1.02–1.07). Functional annotation using data from the Encyclopedia of DNA Elements (ENCODE) project revealed two putative functional variants, rs62331150 and rs73838678 in linkage disequilibrium (LD) with rs9790517 (r2 ≥ 0.90) residing in the active promoter or enhancer, respectively, of the nearest gene, TET2. Both variants are located in DNase I hypersensitivity and transcription factor–binding sites. Using data from both The Cancer Genome Atlas (TCGA) and Molecular Taxonomy of Breast Cancer International Consortium (METABRIC), we showed that rs62331150 was associated with level of expression of TET2 in breast normal and tumor tissue. Conclusion: Our study identified two independent association signals at 4q24 in relation to breast cancer risk and suggested that observed association in this locus may be mediated through the regulation of TET2. Impact: Fine-mapping study with large sample size warranted for identification of independent loci for breast cancer risk. Cancer Epidemiol Biomarkers Prev; 24(11); 1680–91. ©2015 AACR.

Abstract Background: Unnecessary intervention and overtreatment of indolent disease are common challenges in clinical management of prostate cancer. Improved tools to distinguish lethal from indolent disease are critical. Methods: We performed a genome-wide survival analysis of cause-specific death in 24,023 prostate cancer patients (3,513 disease-specific deaths) from the PRACTICAL and BPC3 consortia. Top findings were assessed for replication in a Norwegian cohort (CONOR). Results: We observed no significant association between genetic variants and prostate cancer survival. Conclusions: Common genetic variants with large impact on prostate cancer survival were not observed in this study. Impact: Future studies should be designed for identification of rare variants with large effect sizes or common variants with small effect sizes. Cancer Epidemiol Biomarkers Prev; 24(11); 1796–800. ©2015 AACR.

AbstractBackgroundWe aimed to determine whether patients and providers were satisfied with teledermoscopy consultation for skin lesions.MethodsFrom 2010 to 2011, patients with clinically suspicious lesions were referred for teledermoscopy by internal medicine physicians. Lesions were digitally photographed using a dermatoscope accessory lens. Images were interpreted by an on‐call dermatologist. We conducted same day surveys of providers and patients after the evaluations.ResultsThe survey response rate was 100%. Of the 20 patients surveyed, all agreed that a teledermoscopic consult was convenient and helpful. Nineteen patients (95%) strongly agreed that teledermoscopy potentially was a cost‐saving tool. All providers thought the evaluations were helpful. Ninety‐five percent of providers and patients reported satisfaction with the ease of use of this technology.ConclusionsTeledermoscopy may give primary care providers the ability to consult with dermatologists when seeing patients in remote or medically underserved areas. This can streamline the referral process and decrease the number of referrals for benign lesions.

Abstract KRAB-Associated Protein 1 (KAP1) is a master regulator for genome stability, heterochromatin formation, and target gene silencing. Many studies have suggested that post-translational modification is required for KAP1 to execute pleiotropic effects. Here, we have identified the physical interaction between KAP1 and SIRT1, which is a widely recognized NAD+-dependent deacetylase in DNA damage response. Therefore, the objective of this study was to characterize the functional significance of KAP1 deacetylation in DNA damage repair signaling. In this study, SIRT1 was identified as a major interacting partner of KAP1 via biochemical approaches. Inhibition of SIRT1 activity by shRNA or selective inhibitor was able to upregulate the total acetylation level on KAP1, suggesting that KAP1 is a novel target of SIRT1. The major SIRT1-regulated KAP1 deacetylation sites (Lys266, Lys377, Lys469, and Lys770) were detected by quantitative mass spectrometric analysis. Next, to explore the functional significance of KAP1 deacetylation in NHEJ-mediated repair, KAP1 depleted cells reconstituted with wild-type KAP1 or 4KR acetylation mutant were used in the in vivo end-joining reporter system. Compare to wild-type KAP1, the 4KR mutant presented higher activity in promoting NHEJ efficiency. Furthermore, 4KR mutant showed stronger binding affinity to 53BP1, a crucial NHEJ promoting factor, and stabilized the 53BP1 foci formation in response to DNA damage. In summary, this study unambiguously established KAP1 as a target of SIRT1 and that the deacetylation of KAP1 on Lys266, Lys377, Lys469, and Lys770 stabilizes the formation of 53BP1 foci on sites of double-strand breaks, thereby promotes NHEJ-mediated repair pathway. Citation Format: Yi-Hui Lin, Zhenkun Lou. SIRT1 regulates the function of the KRAB-associated protein 1 (KAP1) in DNA damage response signaling. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4979. doi:10.1158/1538-7445.AM2015-4979

Abstract Background: Recent bioinformatic analyses and high-throughput sequencing efforts have revealed that RNA editing is a widespread post-transcriptional modification that affects non-coding repetitive elements throughout the genome. This modification, which involves the conversion of adenosine into inosine, is mediated by the action of the adenosine deaminases acting on RNA (ADAR) family of enzymes. Based on reports of dysfunctional ADAR expression in epithelial ovarian cancers (EOC), we hypothesized that single nucleotide polymorphisms (SNPs) in five ADAR genes (ADAD1, ADAR1, ADAR2, ADAR3, SND1) may contribute to EOC susceptibility, potentially by altering the expression of corresponding genes in ovarian tissues. Methods: Using directly genotyped and imputed data from 10,891 invasive EOC cases and 21,693 healthy controls of European ancestry represented in the international Collaborative Oncological Gene-environment Study, we evaluated the frequency of 7,133 SNPs in 5 RNA editing genes. Unconditional logistic regression was used to estimate odds ratios (OR) and 95% confidence intervals (CI) between genotypes and case status under a log-additive genetic model, with adjustment for the first five principal components representing European ancestry. Subgroup analysis was conducted for the four main histologic sub-types of EOC. We also aggregated SNP-level data and conducted gene-level analyses using the Admixture Maximum Likelihood (AML) test and the Sequence-Kernel Association test for the combined effect of common and rare variants (SKAT-CR). Results: Association analysis revealed a top risk-associated SNP (OR (95% CI) = 1.39 (1.17-1.64)) in ADAR3 with P = 1.3 x10−4, 17 additional SNPs in ADAR1, ADAR3, or SND1 with P = 10−4, and 51 SNPs with P = 10−3; this observed number of statistically-significant SNPs was greater than expected. When restricting to the 6,500 invasive serous adenocarcinomas, the magnitude of association strengthened for numerous SNPs. Exploratory analysis for the less common histologic subtypes (endometrioid (n = 1,439), mucinous (n = 696), and clear cell (n = 660)) revealed SNP-level associations (P<10−3) unique to each sub-type. Gene-level analyses based on AML and SKAT-CR revealed that ADAR1 was globally associated (P<0.10) with susceptibility to invasive EOC, with P = 0.024 and P = 0.023, respectively. Expression quantitative trait locus analysis is underway to evaluate associations between genotype and gene expression in EOC and precursor tissues using data from The Cancer Genome Atlas Project and other sources. Conclusions: Preliminary data from this large-scale collaboration suggest that germline variation involving RNA editing genes may influence susceptibility to EOC. This novel finding warrants further investigation, as inherited and acquired alterations affecting RNA editing may serve as important biological mechanisms that promote the development of EOC and possibly other malignancies. Citation Format: Jennifer Permuth-Wey, Brett M. Reid, Y. Ann Chen, Hui-Yi Lin, Alvaro Monteiro, Zhihua Chen, Andrew Berchuck, Georgia Chenevix-Trench, Jennifer Doherty, Simon Gayther, Ellen Goode, Edwin Iversen, Leigh Pearce, Paul D. P. Pharoah, Catherine Phelan, Susan Ramus, Mary Anne Rossing, Joellen Schildkraut, Thomas Sellers, on behalf of the Ovarian Cancer Association Consortium. Inherited variants affecting RNA editing may contribute to ovarian cancer susceptibility. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4635. doi:10.1158/1538-7445.AM2015-4635

Abstract Background: Cell-type-specific super enhancers, or large clusters of mediator-rich enhancers, are highly transcribed regions of the genome with key functions in the maintenance of cell identity and control. These regulatory elements are easily perturbed and enriched for disease-associated sequence variation. Identification of disease-associated super-enhancers and investigation of their sequence variants may help identify important, biologically relevant associations previously overlooked by genome-wide studies. Thus, we sought to identify variants within ovarian tissue specific super-enhancers and investigate their association with epithelial ovarian cancer (EOC) susceptibility. Methods: We utilized genotype data from a European population of ∼11,000 invasive EOC cases and 22,000 healthy controls from the Ovarian Cancer Association Consortium. A public catalog of 478 super-enhancers, identified by peak histone H3K27ac ChIP-seq signal in ovarian tissue, was used to identify 72,116 variants located within 344 super-enhancer regions. Unconditional logistic regression (log-additive model) was used to assess individual SNP-EOC risk associations. Additionally, we reviewed associations within 500 kb of super-enhancer regions to identify proximal associated variants. When LD structure indicated, we employed conditional analysis to determine independence of the super-enhancer signal. Gene-EOC risk associations were assessed with the Admixture Maximum Likelihood (AML) test and Sequence-Kernel Association Test (SKAT) for the combined effect of common and rare (MAF<0.01) variants. Models were adjusted for population stratification using the first five principal components. To adjust for multiple comparisons, a false discovery rate of 10% was used. Results: The most significant SNP observed was rs147487657 (p = 7.7×10−6) on locus 2q37.3 in the HDAC4 gene and there were an additional 12 SNPs with p<1.0×10−4, although not significant at 10% FDR. Gene-level analyses identified two super-enhancers significantly associated with EOC risk. An enhancer at the previously reported 2q31 locus, containing HOXD8 and HOXD-AS2, associated (SKAT p = 4.3×10−5; AML p<0.001) with the top SNP located within the 5′UTR of HOXD8 transcription factor (rs847163 (T>C); p<2.08×10−5). Conditional analysis did not identify an independent association within the super-enhancer region. The second EOC-associated super-enhancer identified was at locus 1q32 (SKAT p = 0.00052, AML p = 0.001) containing the CSRP1 gene. There were 6 SNPs with p<0.001 and 16 SNPs with p<0.01, all were inversely associated with risk and consistent with the top hit at rs927901 T>C (OR = 0.94; 95% CI: 0.90-0.97; p = 2.1×10−4). Associations varied by histologic subtype, possibly due to variance in disease relevant super-enhancer cell-type.Conclusion: These findings suggest germline variation within super-enhancer regions may contribute to EOC susceptibility Citation Format: Brett M. Reid, Jennifer Permuth-Wey, Y. Ann Chen, Hui-Yi Lin, Alvaro Monteiro, Zhihua Chen, Andrew Berchuck, Georgia Chenevix-Trench, Jennifer Doherty, Simon Gayther, Ellen L. Goode, Edwin Iversen, Leigh Pearce, Paul Pharoah, Catherine Phelan, Susan Ramus, Mary Anne Rossing, Joellen Schildkraut, Thomas Sellers, on behalf of the Ovarian Cancer Association Consortium. Variants within super-enhancer regulatory elements associate with epithelial ovarian cancer risk. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4634. doi:10.1158/1538-7445.AM2015-4634

Abstract Background: Genome wide association studies (GWAS) have identified 20 loci associated with epithelial ovarian cancer (EOC) risk. Additional EOC risk loci await identification, and biological approaches may represent a useful strategy for overcoming sample size limitations. The ENCODE project recently concluded that only ∼1.2% of the genome encodes proteins, but at least 20% exhibits biological function and over 80% exhibits biochemical indices of function. Post-GWAS follow-up studies have firmly established that some associations are due to inter-individual differences in non-coding RNAs (ncRNAs). To estimate the magnitude of impact of variants in long non-coding RNAs (lncRNAs), we assessed enrichment of EOC-associated SNPs in lncRNA regions by comparing the density of EOC-associated loci between lncRNA regions, non-lncRNA regions, and the whole genome. Methods: The study population included ∼18,000 invasive EOC cases and 34,000 healthy controls of European ancestry from the Ovarian Cancer Association Consortium with GWAS data imputed to 1000 Genomes Project (1KGP) density. Density was compared with two measures: average chromosome length required for one EOC-associated SNP (kb/locus) and average number of tested SNPs containing one EOC-associated SNP (SNPs/locus). Coordinates and annotation for 13,870 lncRNA genes were downloaded from the publicly available GENCODE (v19) database and were used to annotate SNPs from the 1KGP imputed GWAS data, yielding 13,192 lncRNA genes with biallelic variants imputed at rsquare ≥0.25. A SNP was considered EOC-associated if it reached a significance level of P<10−5. Results: Genome-wide, 11.6% of the 15,123,646 tested SNPs fell within lncRNA regions. Of 5,294 EOC-associated SNPs that were identified, 27.7% were located within lncRNAs, a higher frequency than expected (<v:shape style = “WIDTH: 16.5pt; HEIGHT: 18pt” id = _x0000_i1026 equationxml = ‘14pχ2′ type = “#_x0000_t75”> = 9.35×10−22). Additionally, EOC-associated SNPs were located on average every 237.6 kb (or 1204 tested SNPs) in lncRNA regions versus every 657.7 kb (or 3489 SNPS) in non-lncRNA regions and every 541.5 kb (or 2857 SNPs) in the whole genome, clearly demonstrating an enrichment of EOC-associated loci in lncRNA regions. A total of 1,464 lncRNA SNPs from 53 unique lncRNAs associated with EOC risk (P<10−5); 70 of the identified lncRNA SNPs had P-values between 10−20 and 10−32, and 221 had P-values between 10−10 and 10−15, representing 21 unique regions > 500kb apart. Conclusions: These results justify efforts to integrate functional and population level genetic data on lncRNA regions to explore a new paradigm that addresses the problem of the missing heritability for cancer where restrictions in sample size and statistical power limit the ability to discover additional risk loci. Citation Format: Thomas A. Sellers, Brett M. Reid, Y. Ann Chen, Hui-Yi Lin, Edward Richards, Jamie Teer, Alvaro Monteiro, Zhihua Chen, Andrew Berchuck, Georgia Chenevix-Trench, Jennifer Doherty, Ellen Goode, Edwin Iverson, Leigh Pearce, Paul Pharoah, Catherine Phelan, Susan Ramus, Mary Anne Rossing, Joellen Schildkraut, Jin Cheng, Simon Gayther, Jennifer Permuth-Wey, on behalf of the Ovarian Cancer Association Consortium. Evidence that long non-coding RNA variants associate with epithelial ovarian cancer risk. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4633. doi:10.1158/1538-7445.AM2015-4633

Abstract Many germline genetic variants have been found to be associated with susceptibility to epithelial ovarian cancer (EOC), but currently few associations have been identified for EOC outcomes such as overall survival. In the absence of evidence for common germline markers, it is possible that rare variants not captured in GWAS arrays may be associated with overall survival. We hypothesize that rare variants, either individually or combined across a gene, may be associated with overall survival in EOC. We utilized data from two genotyping projects of the Ovarian Cancer Association Consortium (OCAC) that used commercial exome-based arrays. The primary sample, genotyped on the Affymetrix Axiom Exome Array at 227,892 standard and custom variants, consisted of 14 independent EOC studies (4293 cases; 2257 deaths). The secondary sample, genotyped on the Illumina Infinium HumanExome BeadChip at 114,620 variants, consisted of six additional EOC studies (1744 cases; 1027 deaths). Both sets were analyzed using Cox proportional hazards regression to determine the association of each variant with overall survival time. Meta-analysis was conducted across samples at 73,203 overlapping variants. Gene-level tests of over 18,000 genes were also conducted using burden tests and Sequencing Kernel Association Tests (SKAT) adapted for Cox proportional hazards models. Gene-level tests were conducted separately for the primary and secondary cases and then were meta-analyzed. Models were adjusted for age, site, and three principle components. Six individual variants had meta-analysis p-values < 1.0E-4, five of which were rare (MAF < 0.003). Common variant rs8170 (MAF = 0.2) in BABAM1 encoding BRISC and BRCA1 A complex member 1 - previously reported to be associated with EOC risk_showed evidence of association with overall survival (Pmeta = 1.5E-5) and consistent estimates of effect size across both samples (HR1 = 1.18, HR2 = 1.19). In imputed genotype data from over 18,000 subjects from OCAC genotyped on the COGS array, this association reaches genome-wide significance (HR = 1.15, P = 4.7E-9). Five genes had gene-level meta-analysis p-values < 1.0E-4, including top ranked ATG2B (p = 1.1E-6), an autophagy gene important for apoptosis; zinc finger protein PEG3 (p = 1.8E-5), playing a role in autophagy and down-regulated in EOC; helicase-like transcription factor HLTF (p = 1.9E-5), involved in post-replication DNA repair; mitochondrial fission regulator 1-like gene FAM54B (p = 3.1E-5); and T-cell surface glycoprotein CD1E (p = 8.8E-5). With over 6000 EOC cases in the primary and secondary samples, this is the largest genome-wide assessment of rare variant genetic associations with EOC overall survival. Variant rs8170 in BABAM1, a necessary component of the BRCA1 complex, is associated with overall survival. Single variant and gene-level analyses provide complementary approaches, suggesting additional candidates that warrant follow-up study. Citation Format: Stacey J. Winham, Brooke L. Fridley, Melissa C. Larson, Zachary Fogarty, Andrew Berchuck, Yian Ann Chen, Hui-Yi Lin, Georgia Chenevix-Trench, Jenny Permuth-Wey, Thomas A. Sellers, Ailith Pirie, Ellen L. Goode, Ovarian Cancer Association Consortium. Investigation of exome variants associated with overall survival in ovarian cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4636. doi:10.1158/1538-7445.AM2015-4636