Vierkant, Robert A.

Background and Purpose —Previous studies have shown that the volume of intracerebral hemorrhage and Glasgow Coma Score (GCS) on admission are powerful predictors of 30-day mortality. However, the significance of hydrocephalus associated with deep cerebral hemorrhage has not been studied extensively. The purpose of this study was to determine the prognostic indicators of 30-day mortality in patients with deep cerebral hemorrhage. Methods —We studied 100 consecutive patients with deep cerebral hemorrhage between 1994 and 1998. Deep cerebral hemorrhage was divided into 2 groups: putaminal hemorrhage (lateral group) and thalamic and caudate hemorrhage (medial group). Univariate and multivariate logistic regression analyses were performed to determine independent prognostic indicators of 30-day mortality. Results —Hydrocephalus was present in 40 of the 100 patients. The 30-day mortality was 29%, and hydrocephalus was present in 76% of those who died. Multivariate analyses showed 2 independent prognostic indicators of 30-day mortality for putaminal hemorrhage: GCS ≤8 ( P =0.002, odds ratio [OR] 37.7, CI 3.6 to 396.9) and hydrocephalus ( P =0.005, OR 27.4, CI 2.7 to 282.6). However, only GCS ≤8 ( P =0.0002, OR 16.5, CI 3.7 to 73.4) was predictive of 30-day mortality for thalamic and caudate hemorrhage. This model (GCS ≤8 and hydrocephalus) has a sensitivity of 57% and a specificity of 91% for predicting 30-day mortality for putaminal hemorrhage. When both attributes were present in putaminal hemorrhage (GCS ≤8 and hydrocephalus), 1 (11%) of 9 patients survived, and when both attributes were missing 28 (100%) of 28 patients survived. Conclusions —Obstructive hydrocephalus on admission in a comatose patient with a putaminal hemorrhage predicts 30-day mortality.

Abstract Background: Mitochondria contribute to oxidative stress, a phenomenon implicated in ovarian carcinogenesis. We hypothesized that inherited variants in mitochondrial-related genes influence epithelial ovarian cancer (EOC) susceptibility. Methods: Through a multicenter study of 1,815 Caucasian EOC cases and 1,900 controls, we investigated associations between EOC risk and 128 single nucleotide polymorphisms (SNPs) from 22 genes/regions within the mitochondrial genome (mtDNA) and 2,839 nuclear-encoded SNPs localized to 138 genes involved in mitochondrial biogenesis (BIO, n = 35), steroid hormone metabolism (HOR, n = 13), and oxidative phosphorylation (OXP, n = 90) pathways. Unconditional logistic regression was used to estimate OR and 95% CI between genotype and case status. Overall significance of each gene and pathway was evaluated by using Fisher's method to combine SNP-level evidence. At the SNP level, we investigated whether lifetime ovulation, hormone replacement therapy (HRT), and cigarette smoking were confounders or modifiers of associations. Results: Interindividual variation involving BIO was most strongly associated with EOC risk (empirical P = 0.050), especially for NRF1, MTERF, PPARGC1A, ESRRA, and CAMK2D. Several SNP-level associations strengthened after adjustment for nongenetic factors, particularly for MTERF. Statistical interactions with cigarette smoking and HRT use were observed with MTERF and CAMK2D SNPs, respectively. Overall variation within mtDNA, HOR, and OXP was not statistically significant (empirical P > 0.10). Conclusion: We provide novel evidence to suggest that variants in mitochondrial biogenesis genes may influence EOC susceptibility. Impact: A deeper understanding of the complex mechanisms implicated in mitochondrial biogenesis and oxidative stress may aid in developing strategies to reduce morbidity and mortality from EOC. Cancer Epidemiol Biomarkers Prev; 20(6); 1131–45. ©2011 AACR.

Abstract Defective microRNA (miRNA) biogenesis contributes to the development and progression of epithelial ovarian cancer (EOC). In this study, we examined the hypothesis that single nucleotide polymorphisms (SNP) in miRNA biogenesis genes may influence EOC risk. In an initial investigation, 318 SNPs in 18 genes were evaluated among 1,815 EOC cases and 1,900 controls, followed up by a replicative joint meta-analysis of data from an additional 2,172 cases and 3,052 controls. Of 23 SNPs from 9 genes associated with risk (empirical P < 0.05) in the initial investigation, the meta-analysis replicated 6 SNPs from the DROSHA, FMR1, LIN28, and LIN28B genes, including rs12194974 (G>A), an SNP in a putative transcription factor binding site in the LIN28B promoter region (summary OR = 0.90, 95% CI: 0.82–0.98; P = 0.015) which has been recently implicated in age of menarche and other phenotypes. Consistent with reports that LIN28B overexpression in EOC contributes to tumorigenesis by repressing tumor suppressor let-7 expression, we provide data from luciferase reporter assays and quantitative RT-PCR to suggest that the inverse association among rs12194974 A allele carriers may be because of reduced LIN28B expression. Our findings suggest that variants in LIN28B and possibly other miRNA biogenesis genes may influence EOC susceptibility. Cancer Res; 71(11); 3896–903. ©2011 AACR.

Abstract INTRODUCTION: The centrosome is the primary microtubule organizing center of the cell. Numerical or functional defects of the centrosome can result in improper chromosome segregation, leading to aneuploidy and polyploidy. Because many cancer cells have aberrant numbers of chromosomes, many have suggested a link between centrosomes and cancer. We conducted an association study to determine whether common genetic variations in 100 genes involved in maintaining and regulating centrosome structure and function account in part for the contribution of the pathway to breast cancer risk. METHODS: Incident cases of BC were identified at the Mayo Clinic in Rochester, MN from 2001 through 2005. Controls (N=843) were frequency matched to cases (N=798) on age and region of residence. Seven hundred-eighty-two tagging and candidate functional SNPs were selected from 100 genes involved in the structure and/or function of the centrosome. Genotyping was performed on an Illumina BeadLab using the Illumina GoldenGate genotyping assay. Logistic regression was used to calculate odds ratios and 95% confidence intervals for BC. Women having two copies of the most common allele were defined as the referent category. Linear regression was used to estimate mean and 95% CI for PD for each additional copy of the variant allele. All models were adjusted for age, residence, and confounding factors. RESULTS: Forty-eight SNPs from 29 genes (out of 782 SNPs examined) showed evidence of significant associations with breast cancer risk in our population in the log-additive (1 degree of freedom (d.f.)) model (Ptrend< 0.05). One SNP, rs1374468 SNP in TACC3, displayed the most significant association with risk in the overall analyses (Ptrend = 0.001). Two SNPs from each of seven genes (AJUBA, CHUK, MCPH1, NEK7, PAK1, PIK3CB, and PINS) were significantly associated with risk of breast cancer (Ptrend < 0.05). Three genes, AXIN2, NIN, NUMA1, had four or more SNPs that were significantly associated with breast cancer risk (Ptrend < 0.05). All of the SNPs in these four genes were in strong linkage disequilibrium (r2>0.6). No single SNP association was significant after adjustment for multiple comparisons via permutation testing. Haplotype analyses showed 20 genes for which global haplotypes were significantly associated with breast cancer risk. Of these, haplotype blocks in six genes (14-3-3 Epsilon, CDC16, CD-TOG, KIF2, NEK9, NPM2) were significantly associated with risk (global P<0.05), although none of the individual SNPs in these six genes displayed significant associations. This suggests that the associations of these specific combinations of alleles with risk were not due to the effects of individual SNPs. CONCLUSION: These analyses provide some evidence to support an association between genetic variation in centrosomal genes and risk of breast cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2852.

Abstract Mitochondria have been implicated in carcinogenesis because of their central role in apoptosis, free radical production, and cellular energy metabolism. Although studies have identified somatic mitochondrial DNA mutations in ovarian cancers, little attention has been directed to the possible role of germline mitochondrial variants in mitochondrial dysfunction and subsequent ovarian cancer development. Through an ongoing epithelial ovarian cancer genome-wide association study of 1,856 invasive cases (59% with serous carcinomas) and 1,912 controls (frequency-matched on age, race, and study site), we genotyped 138 SNPs tagging variation in the maternally-inherited mitochondrial genome (mtDNA) to evaluate the hypothesis that single nucleotide polymorphisms (SNPs) in mtDNA are associated with ovarian cancer risk. All subjects were non-Hispanic, non-Jewish Caucasians, and were genotyped with the Illumina 610-quad Beadchip. Samples and SNPs with call rates less than 95% were excluded. Subjects with ambiguous gender, unresolved identical genotypes, and less than 80% European ancestry were excluded. For each SNP, logistic regression was used to estimate odds ratios (OR) and corresponding 95% confidence intervals (CI) between carriers of the minor versus major maternally-inherited allele and case status. Subgroup analysis was conducted to estimate allele-specific risks between serous-only cases and controls. Haplotype analysis was performed for regions containing statistically significant SNPs (p<0.05). Two rare SNPs, C16329T in the displacement loop gene (MT-D-loop) (OR: 6.72, 95%CI: 1.5-29.8, p=0.004; minor allele frequency (MAF)=0.001) and C15905T in the tRNA threonine gene (MT-TT) (OR: 1.60, 95%CI: 1.1-2.3, p=0.009; MAF=0.027), were associated with increased ovarian cancer risk; whereas T6777C in the cytochrome c oxidase subunit 1 gene (MT-CO1) (OR: 0.68, 95%CI: 0.51-0.91, p=0.009; MAF=0.063) was associated with decreased risk. All three SNPs remained statistically significant after adjustment for multiple comparisons within the mitochondrial SNPs using permutation testing. C16329T and T6777C were also significantly associated among cases with serous histology, with ORs (95% CIs) of 6.13 (1.2-29.6) and 0.61 (0.43-0.87), respectively. Haplotype analysis for each of the three regions investigated revealed significant haplotype-disease associations; however, findings suggest that the respective associations were driven by the most significant SNP(s) listed above. This is the first large-scale epidemiologic study to provide evidence that SNPs in mitochondrial genes may be novel risk factors for ovarian cancer. Future investigation of additional mitochondrial variants (i.e. nuclear-encoded mitochondrial proteins) is necessary to more comprehensively investigate this association. Furthermore, replication of the most promising SNPs in a larger population is warranted to validate these findings. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2835.

Abstract As the leading cause of mortality among gynecologic cancers, it is critical to increase understanding of ovarian cancer pathogenesis, which remains largely unknown. Several xenobiotic metabolizing enzymes are responsible for processing and inactivating carcinogens that could initiate ovarian carcinogenesis. Here, we hypothesize that single-nucleotide polymorphisms (SNPs) in genes encoding xenobiotic metabolizing enzymes may be associated with differential risk of ovarian cancer. Cases with epithelial ovarian cancer (N=930) and controls (N=1,037), frequency-matched on age and geographic residence, were enrolled at Mayo Clinic and Duke University, and genotyped at 163 tagSNPs in EPHX1, SULT1E1, ADH5, ADH4, ADH6, ADH1A, ADH1B, ADH1C, NQO2, NAT1, NAT2, GSTP1, CYP19A1, CYP1A1, CYP1A2, and NQO1 using an Illumina GoldenGate™ BeadArray array. Logistic regression was used to estimate odds ratios (ORs) and adjust for population structure and other potential confounders, including oral contraceptive use, parity, and family history. SNP-specific statistical analyses identified several noteworthy results including an association with increased risk of ovarian cancer with a SNP (rs2917666) in NAD(P)H dehydrogenase, quinone 1 (NQO1) (per-allele OR 1.16, 95% confidence interval (CI) 1.01-1.33, p=0.04). The SNP rs1051740 in epoxide hydrolase 1, microsomal (xenobiotic) (EPHX1) was suggestive of increased risk of invasive serous disease (TT v CT OR 0.85, 95% CI 0.66-1.09, TT v CC 1.29, 95% CI 0.89-1.88, p=0.09). In an effort to replicate these observations, a subset of 11 SNPs with a p-value < 0.10 were then genotyped in 727 invasive cancer cases and 1,065 controls from an Australian ovarian cancer study using a Sequenom iPlex assay. Results of combined analyses confirmed an association between EPHX1 rs1051740 and increased invasive serous ovarian cancer risk (TT v CT OR 0.99, 95% CI 0.77-1.29, TT v CC OR 1.61, 95% CI 1.23-2.11, p=0.001; recessive OR 1.61, 95% CI 1.25-2.09, p=0.001). This gene is involved in the activation and detoxification of tobacco-derived carcinogens, and smoking has been associated with the mucinous histologic subtype of ovarian cancer. Analyses by histological subtype and gene-environment interaction analyses considering genotype in conjunction with tobacco use and alcohol intake are underway. Due to the important role of EPHX1 in metabolizing pro-carcinogens and the observed association between rs1051740 and risk of invasive serous disease, an examination of this SNP in a larger ovarian cancer consortium is warranted. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 914.

Abstract Using data from patients with invasive epithelial ovarian cancer, we sought to examine whether overall survival was associated with common inherited variation in candidate genes and genomic regions including those involved in angiogenesis, mitosis, and regions of interest from genome-wide association and expression studies. We conducted a two-stage analysis based on Mayo Clinic patients with replication of peak results in the Cancer Genome Atlas (TCGA). Ovarian cancer patients (N=312) seen at the Mayo Clinic diagnosed from 1999 to 2006 with median follow-up of 3.7 years (range, 0.1 − 9.5 years) and 172 observed deaths were analyzed at 993 single nucleotide polymorphisms (SNPs) in 179 genes. For all patients and for serous patients only (N=192 with 129 observed deaths), Cox regression was used to estimate hazard ratios (HRs) and 95% confidence intervals (CIs) for each SNP, adjusted for other prognostic factors. Twenty-nine SNPs in ten genes revealed p < 0.01 among all cases or among serous cases only, including 21 independent SNP at r2=0.9; analyses within four genes (HGF, PLG, PRKACB, and DCTN5) yielded p < 0.001. The strongest result was in HGF at rs1800793 (all cases: HR 1.7, 95% CI 1.3 − 2.2, p-trend 1.9 E-5; serous cases: HR 1.6, 95% CI 1.2 − 2.2, p-trend 1.8 E-3). A correlated SNP rs2214825 also showed association (r2=0.76; all cases: HR 1.4, 95% CI 1.1 − 1.8, p-trend 3.3 E-3; serous cases: HR 1.4, 95% CI 1.0 − 1.9, p-trend 0.03), and PCA gene-level tests of the HGF locus were also significantly associated with survival (all cases, p=3.7 E-4; serous cases, p=0.01). Among all cases, differential survival was also associated with DCTN5 rs12447304 (HR 1.8, 95% CI 1.3-2.6; p=7.1 E-4), and among serous cases, with PLG rs783173 (HR 1.6, 95% CI 1.2 − 2.0, p = 8.1 E-4) and PRKACB rs1402694 (HR 1.7, 95% CI 1.3 − 2.2, p=3.2 E-4). We then examined SNPs in the most suggestive gene, HGF, using data from TCGA which had genotyped four overlapping HGF SNPs in 350 invasive cases (188 deaths). Covariate-adjusted TCGA analysis at rs2214825 revealed consistent results (HR 1.7, 95% CI 1.3-2.2, p=1.4 E-4) as did analyses of three other HGF SNPs (HR range 1.3 to 1.5, p range 3.4 E-3 to 0.049); correlation between SNP genotypes and HGF expression was also observed (p=1.4 E-3, rs2214825, probe 210997_at). While this is the first report of SNPs in HGF, it has long been known that the HGF signaling pathway, activated by HGF ligation of c-MET, plays a key role ovarian cancer cell growth, migration, and invasion. c-MET is overexpressed in 40%-60% of ovarian tumors, and high overexpression is associated with lower overall survival. In addition, HGF has been shown to enhance the response of ovarian cancer cells to low doses of paclitaxel and cisplatin by activating the intrinsic apoptotic pathway, leading to cancer cell death. These results now suggest that inherited differences in HGF among women with invasive disease may play a role in survival, holding potential clues for tailored avenues of treatment. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 897.

Abstract Recent GWAS have identified and confirmed associations between several SNPs in the gene desert located on chromosome 8q24 with risk of cancers of the breast, prostate, glioma, bladder and colorectum. Evidence supporting an association with ovarian cancer is limited and inconsistent. Here we evaluate preliminary results in the 8q24 region from stage I of an ongoing invasive epithelial ovarian cancer GWAS among 1864 cases (1096 serous cases) and 1912 controls. All subjects were self-reported non-Hispanic non-Jewish Caucasian, and were genotyped with the Illumina 610quad Chip. Samples and SNPs with call rates less than 95% were excluded (9.6% and 9.9%, respectively) as well as 128 subjects with ambiguous gender, unresolved identical genotypes and less than 80% European ancestry were also excluded. Our most significant hit in the region was rs6651252 located at 8q24.21 with p = 0.000125 for all ovarian cancer, and 1.02*10−5 for serous cases only. This region is located approximately 849 kb downstream of the previous 8q24.21 findings on other cancers, and was not in linkage disequilibrium with any of the haplotype blocks reported previously. We further examined the block structure of this region with eight genotyped SNPs located within 129.61 to 129.64 MB (rs10088218, rs1516973, rs1516975, rs16903097, rs10098821, rs6651252, rs1561928, and rs1561927) and performed haplotype analysis to evaluate the association with ovarian cancer risk. One haplotype (AAGCAGGG, estimated frequency=0.11) was significantly associated with decreased ovarian cancer risk with an odds ratio of 0.67 (all cases, 95% CI: 0.78-0.91, p=0.0026, empirical p= 0.0020). This region contains no known genes, but it is located upstream and overlaps with a known copy number variation (CNV) reported by Redon et al. (2006). Previous published literature on array CGH and CNV results from TCGA (The Cancer Genome Atlas) also indicate that the 8q24.21 region is associated with frequent gains/amplification in ovarian tumors. Combined analysis of SNPs and CNV identified in this region is underway to elucidate the joint effect of these two types of genetic variation on ovarian cancer risk. Our result indicates a novel locus on 8q24.21 is associated with epithelial ovarian cancer susceptibility. Haplotype analysis reveals one specific haplotype that is associated with decreased ovarian cancer risk compared to the most common haplotype. Given previous 8q24 findings from other cancers and evidence from publicly available resources, fine-mapping and resequencing of this region is warranted. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4724.

Abstract Background: We previously identified a panel of genes associated with outcome of ovarian cancer. The purpose of the current study was to assess whether variants in these genes correlated with ovarian cancer risk. Methods and Findings: Women with and without invasive ovarian cancer (749 cases, 1,041 controls) were genotyped at 136 single nucleotide polymorphisms (SNPs) within 13 candidate genes. Risk was estimated for each SNP and for overall variation within each gene. At the gene-level, variation within MSL1 (male-specific lethal-1 homolog) was associated with risk of serous cancer (p=0.03); haplotypes within PRPF31 (PRP31 pre-mRNA processing factor 31 homolog) were associated with risk of invasive disease (p=0.03). MSL1 rs7211770 was associated with decreased risk of serous disease (OR 0.81, 95% CI 0.66-0.98; p=0.03). SNPs in MFSD7, BTN3A3, ZNF200, PTPRS, and CCND1A were inversely associated with risk (p<0.05), and there was increased risk at HEXIM1 rs1053578 (p=0.04, OR 1.40, 95% CI 1.02-1.91). Conclusions: Tumor studies can reveal novel genes worthy of follow-up for cancer susceptibility. Here, we found that inherited markers in the gene encoding MSL1, part of a complex that modifies the histone H4, may decrease risk of invasive serous ovarian cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4718.

Abstract Genome-wide association analyses have identified an ovarian cancer susceptibility locus near BNC2 at 9p22.2 (OR 0.82, p 5.1 × 10−19, Nat Genet 41:996-1000, 2009), based on 1,772 and 2,354 Stage I cases and controls, 4,803 and 5,237 Stage 2 cases and controls, and extensive Stage 3 analysis in independent samples. Here, we report on Stage 3 analysis of nine additional regions with Stage 2 results at p < 10−4. Stage 1 samples were genotyped using the Illumina 610Quad and imputed at up to 2 million SNPs; Stage 2 samples were genotyped on an Illumina iSelect of 22,790 SNPs. Stage 3 samples from 4,335 cases (including 2,197 serous subtype) and 5,951 controls (14 studies) were genotyped using Illumina or Sequenom at up to 32 SNPs on chromosomes 1, 2, 3, 7, 8, 11, and 17. No evidence of heterogeneity by study site was observed. Logistic regression revealed four regions with ordinal p ≤ 10−7 in all cases or serous subtype only. The most significant finding for all cases was for rs2072590 at 2q31.1 (all cases OR 1.15, p=8.1 × 10−13; serous only OR 1.19 p=9.0 × 10−13) which is located 3.8 kb 3-prime of HOXD3, a homeobox gene which also had an intronic SNP (rs2301301) with p=3.1 × 10−5 (all cases). Second, we observed a locus at 8q24.21 highlighted by rs10088218 (all cases OR 0.84, p=1.3 × 10−9; serous only OR 0.75 p=2.2 × 10−14). This locus resides on the opposite side of MYC and 1.3 Mb from the 8q24 regions harboring loci for other cancers, and is independent of the other cancer-associated variants (r2 < 0.02). Third, at 3q25.31, promising results were observed at rs2665390, a SNP intronic to TIPARP (TiPARP, TCDD-inducible poly(ADP-ribose) polymerase; all cases OR 1.19, p=8.7 × 10−7; serous only OR 1.26, p=3.2 × 10−8). Finally, at 17q21.32, rs9303542 was associated with risk particularly among serous cases (all cases OR 1.13, p=1.5 × 10−6; serous only OR 1.16 p=4.3 × 10−7); this SNP is intronic to SKAP1 (src kinase associated phosphoprotein 1). We evaluated the functional role of genes at these loci in ovarian cancer and normal ovarian epithelial cells and in an in vitro model of neoplastic initiation and progression from normal ovarian epithelial cells after CYMC and KRAS activation. Both BNC2 and TiPARP were down regulated in cancer versus normal cells (p=5 × 10−9 and p=3.4 × 10−5, respectively) suggesting that loss of function is required for cancer development. Over-expressing CMYC (at 8q24) induced neoplastic transformation of normal ovarian cells. Both BNC2 and TiPARP were down-regulated following CMYC over-expression and showed significant loss of function with further transformation after mutant KRAS activation (p=7.8 × 10−7 and p=6.3 × 10−4, respectively). These results emphasize the importance of genome-wide association studies and consortia for the identification of moderate-risk alleles in rare cancers. Importantly, our findings unravel the genetic components of this disease and suggest functionally-significant novel targets. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3860.