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Purpose: To determine if gene expression signature of invasive oral squamous cell carcinoma (OSCC) can subclassify OSCC based on survival.Experimental Design: We analyzed the expression of 131 genes in 119 OSCC, 35 normal, and 17 dysplastic mucosa to identify cluster-defined subgroups. Multivariate Cox regression was used to estimate the association between gene expression and survival. By stepwise Cox regression, the top predictive models of OSCC-specific survival were determined and compared by receiver operating characteristic analysis.Results: The 3-year overall mean ± SE survival for a cluster of 45 OSCC patients was 38.7 ± 0.09% compared with 69.1 ± 0.08% for the remaining patients. Multivariate analysis adjusted for age, sex, and stage showed that the 45 OSCC patient cluster had worse overall and OSCC-specific survival (hazard ratio, 3.31; 95% confidence interval, 1.66-6.58 and hazard ratio, 5.43; 95% confidence interval, 2.32-12.73, respectively). Stepwise Cox regression on the 131 probe sets revealed that a model with a term for LAMC2 (laminin γ2) gene expression best identified patients with worst OSCC-specific survival. We fit a Cox model with a term for a principal component analysis-derived risk score marker and two other models that combined stage with either LAMC2 or PCA. The area under the curve for models combining stage with either LAMC2 or PCA was 0.80 or 0.82, respectively, compared with 0.70 for stage alone (P = 0.013 and 0.008, respectively).Conclusions: Gene expression and stage combined predict survival of OSCC patients better than stage alone.

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Supplementary Tables 1-3 from A Sex-Specific Association between a 15q25 Variant and Upper Aerodigestive Tract Cancers

Background: The matrix metalloproteinases (MMP) cause degradation of the extracellular matrix and basement membranes, and thus may play a key role in cancer development.Methods: In our search for biomarkers for oral squamous cell carcinomas (OSCC), we compared primary OSCC, oral dysplasia and control subjects with respect to: (i) expression of MMP1, MMP3, MMP10, and MMP12 in oral epithelial tissue using Affymetrix U133 2.0 Plus GeneChip arrays, followed by quantitative reverse transcription-PCR (qRT-PCR) for MMP1, and (ii) determination of MMP1 and MMP3 concentrations in saliva.Results:MMP1 expression in primary OSCC (n = 119) was >200-fold higher (P = 7.16 × 10−40) compared with expression levels in nonneoplastic oral epithelium from controls (n = 35). qRT-PCR results on 30 cases and 22 controls confirmed this substantial differential expression. The exceptional discriminatory power to separate OSCC from controls was validated in two independent testing sets (AUC% = 100; 95% CI: 100–100 and AUC% = 98.4; 95% CI: 95.6–100). Salivary concentrations of MMP1 and MMP3 in OSCC patients (33 stage I/II, 26 stage III/IV) were 6.2 times (95% CI: 3.32–11.73) and 14.8 times (95% CI: 6.75–32.56) higher, respectively, than in controls, and displayed an increasing trend with higher stage disease.Conclusion: Tumor and salivary MMPs are robust diagnostic biomarkers of OSCC.Impact: The capacity of MMP gene expression to identify OSCC provides support for further investigation into MMPs as potential markers for OSCC development. Detection of MMP proteins in saliva in particular may provide a promising means to detect and monitor OSCC noninvasively. Cancer Epidemiol Biomarkers Prev; 20(12); 2628–36. ©2011 AACR.

Association of frequency of marijuana use, duration of marijuana, and joint years of marijuana use with oropharyngeal and oral tongue cancers

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Description of the studies utilized in assessment of the association of marijuana use and head and neck cancer in INHANCE

Association of Ever Marijuana use and oropharyngeal cancer stratified by HPV 16 L1 serostatus.

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Purpose: To determine the differential gene expression between oral squamous cell carcinoma (OSCC) with and without metastasis to cervical lymph nodes and to assess prediction of nodal metastasis by using molecular features.Experimental Design: We used Affymetrix U133 2.0 plus arrays to compare the tumor genome-wide gene expression of 73 node-positive OSCCs with 40 node-negative OSCCs (≥18 months). Multivariate linear regression was used to estimate the association between gene expression and nodal metastasis. Stepwise logistic regression and receiver operating characteristics (ROC) analysis were used to generate predictive models and to compare these with models by using tumor size alone.Results: We identified five genes differentially expressed between node-positive and node-negative OSCCs after adjusting for tumor size and human papillomavirus status: REEP1, RNF145, CTONG2002744, MYO5A, and FBXO32. Stepwise regression identified a four-gene model (MYO5A, RFN145, FBXO32, and CTONG2002744) as the most predictive of nodal metastasis. A leave-one-out ROC analysis revealed that our model had a higher area under the curve (AUC) for identifying occult nodal metastasis compared with that of a model by tumor size alone (respective AUC: 0.85 and 0.61; P = 0.011). A model combining tumor size and gene expression did not further improve the prediction of occult metastasis. Independent validation using 31 metastatic and 13 nonmetastatic cases revealed a significant underexpression of CTONG2002744 (P = 0.0004).Conclusions: These results suggest that our gene expression markers of OSCC metastasis hold promise for improving current clinical practice. Confirmation by others and functional studies of CTONG2002744 is warranted. Clin Cancer Res; 17(8); 2466–73. ©2011 AACR.

Association of marijuana use and oral tongue†cancer stratified smoking/drinking intensity

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Background: A history of diabetes is associated with an increased risk of several types of cancers. Whether diabetes is a risk factor for head and neck cancer (HNC) has received little attention.Methods: We pooled data from 12 case–control studies including 6,448 cases and 13,747 controls, and estimated OR and 95% CI for the associations between diabetes and HNC, adjusted for age, education level, sex, race/ethnicity, study center, cigarette smoking, alcohol use, and body mass index.Results: We observed a weak association between diabetes and the incidence of HNC overall (OR, 1.09; 95% CI: 0.95–1.24). However, we observed a modest association among never smokers (OR, 1.59; 95% CI: 1.22–2.07), and no association among ever smokers (OR, 0.96; 95% CI: 0.83–1.11); likelihood ratio test for interaction P = 0.001.Conclusion: A history of diabetes was weakly associated with HNC overall, but we observed evidence of effect modification by smoking status, with a positive association among those who never smoked cigarettes.Impact: This study suggests that glucose metabolism abnormalities may be a HNC risk factor in subgroups of the population. Prospective studies incorporating biomarkers are needed to improve our understanding of the relationship between diabetes and HNC risk, possibly providing new strategies in the prevention of HNC. Cancer Epidemiol Biomarkers Prev; 21(2); 294–304. ©2011 AACR.

Age and Sex stratified analysis on the association of ever marijuana use and oral tongue cancer

Background: The incidence of oropharyngeal and oral tongue cancers has increased over the last 20 years which parallels increased use of marijuana among individuals born after 1950.Methods: A pooled analysis was conducted comprising individual-level data from nine case–control studies from the United States and Latin America in the INHANCE consortium. Self-reported information on marijuana smoking, demographic, and behavioral factors was obtained from 1,921 oropharyngeal cases, 356 oral tongue cases, and 7,639 controls.Results: Compared with never marijuana smokers, ever marijuana smokers had an elevated risk of oropharyngeal [adjusted OR (aOR), 1.24; 95% confidence interval (CI): 1.06–1.47] and a reduced risk of oral tongue cancer (aOR, 0.47; 95% CI, 0.29, 0.75). The risk of oropharyngeal cancer remained elevated among never tobacco and alcohol users. The risk of oral tongue cancer was reduced among never users of tobacco and alcohol. Sensitivity analysis adjusting for potential confounding by HPV exposure attenuated the association of marijuana use with oropharyngeal cancer (aOR, 0.99; 95% CI, 0.71–1.25), but had no effect on the oral tongue cancer association.Conclusions: These results suggest that the association of marijuana use with head and neck carcinoma may differ by tumor site.Impact: The associations of marijuana use with oropharyngeal and oral tongue cancer are consistent with both possible pro- and anticarcinogenic effects of cannabinoids. Additional work is needed to rule out various sources of bias, including residual confounding by HPV infection and misclassification of marijuana exposure. Cancer Epidemiol Biomarkers Prev; 23(1); 160–71. ©2013 AACR.

Description for sensitivity analysis to adjust for potential unmeasured confounding

Supplementary Figure S1; Supplementary Tables S1-S4.

Sensitivity analysis of the effect of misclassification of marijuana use exposure on the univariate association of ever marijuana use with oropharyngeal cancer

Supplementary Data from A Genetic Expression Profile Associated with Oral Cancer Identifies a Group of Patients at High Risk of Poor Survival

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Background: Sequence variants located at 15q25 have been associated with lung cancer and propensity to smoke. We recently reported an association between rs16969968 and risk of upper aerodigestive tract (UADT) cancers (oral cavity, oropharynx, hypopharynx, larynx, and esophagus) in women (OR = 1.24, P = 0.003) with little effect in men (OR = 1.04, P = 0.35).Methods: In a coordinated genotyping study within the International Head and Neck Cancer Epidemiology (INHANCE) consortium, we have sought to replicate these findings in an additional 4,604 cases and 6,239 controls from 10 independent UADT cancer case–control studies.Results: rs16969968 was again associated with UADT cancers in women (OR = 1.21, 95% CI = 1.08–1.36, P = 0.001) and a similar lack of observed effect in men [OR = 1.02, 95% CI = 0.95–1.09, P = 0.66; P-heterogeneity (Phet) = 0.01]. In a pooled analysis of the original and current studies, totaling 8,572 UADT cancer cases and 11,558 controls, the association was observed among females (OR = 1.22, 95% CI = 1.12–1.34, P = 7 × 10−6) but not males (OR = 1.02, 95% CI = 0.97–1.08, P = 0.35; Phet = 6 × 10−4). There was little evidence for a sexdifference in the association between this variant and cigarettes smoked per day, with male and female rs16969968 variant carriers smoking approximately the same amount more in the 11,991 ever smokers in the pooled analysis of the 14 studies (Phet = 0.86).Conclusions: This study has confirmed a sex difference in the association between the 15q25 variant rs16969968 and UADT cancers.Impact: Further research is warranted to elucidate the mechanisms underlying these observations. Cancer Epidemiol Biomarkers Prev; 20(4); 658–64. ©2011 AACR.

Background: The Fred Hutch/University of Washington Cancer Consortium (Consortium) is a National Cancer Institute-designated comprehensive cancer center located in the Seattle metro area. The Consortium comprises four partnering institutions (Fred Hutchinson Cancer Research Center, University of Washington, Seattle Children's Hospital, and Seattle Cancer Care Alliance [SCCA]) that together address the full spectrum of cancer research and interventions to reduce the burden of cancer among catchment area (CA) residents. The Consortium's CA is thirteen counties in western Washington (WA). In an effort to understand and document ongoing research conducted to address the cancer burden, risk factors, incidence, mortality, and inequities in the CA, we initiated a comprehensive needs assessment. Methods: We conducted a quantitative and qualitative needs assessment from September 2017 to June 2018. Quantitative assessment included secondary data analyses from multiple sources (Seattle-Puget Sound Surveillance, Epidemiology and End Results cancer registry, Behavioral Risk Factor Surveillance Survey data, and WA State Department of Health data). Qualitative assessment included semistructured interviews (n=32) conducted in person and over the telephone with key informants representing county health departments, clinic/health systems, SCCA network sites, and community-based organizations. Interviews included questions about barriers and facilitators to cancer screening, referrals, services, access to care, and cancer inequities. Results: The quantitative data analysis identified the top five cancer incidence sites in the CA as breast, prostate, lung, hematologic malignancies and colorectal; the top five mortality sites in the CA are lung, prostate, breast, hematologic malignancies and colorectal. Comparisons of incidence and mortality for the region revealed several inequities: 1) American Indians and Alaska Natives (AIAN) had the most numerous and severe inequities for incidence and mortality of the top cancers; 2) non-Hispanic Blacks had the most severe mortality for prostate cancer; and 3) outlying rural counties had higher incidence and mortality rates than urban areas. Through the qualitative data analysis, we identified seven major themes that influenced cancer prevention and control, including geography, social determinants of health, financial issues, issues around provider trust and communication, issues with agency partnerships/collaboration, and race/ethnicity. Qualitative interviews suggested opportunities for the Consortium to address barriers that underlie several major cancer inequities in the CA. Conclusion: The Consortium will use the needs assessment as a baseline measure to evaluate future research that addresses the cancer burden in the CA. The Consortium now has information to take an informed approach to engage key stakeholders to collaborate and holistically improve cancer research and outcomes for all CA residents. Citation Format: Katherine J. Briant, Beti Thompson, David R. Doody, Clara Reyes, Stephen M. Schwartz, Peggy A. Hannon, Jason A. Mendoza. Collecting evidence to drive research that addresses community cancer needs in Western Washington [abstract]. In: Proceedings of the Eleventh AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2018 Nov 2-5; New Orleans, LA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2020;29(6 Suppl):Abstract nr C004.

Background: Existing comorbidity indices were not developed for adolescent and young adults (AYA) 15 to 39 years of age. The aim of this study was to assess impact of comorbidities on health care service needs and health status among AYA cancer survivors using the newly developed AYA HOPE comorbidity index in comparison with the existing indices. Methods: Data on comorbid conditions were obtained from medical records and service needs and health status were from a survey of AYA cancer survivors. Prevalence of comorbidities was based on the AYA HOPE index. Charlson and NCI indices were compared. Multivariable logistic regression was used. Results: Of the 485 patients, 14.6% had ≥2 comorbidities based on the AYA HOPE Index. Prevalence of mental illness and obesity/overweight, which were not included in existing indices, were 8.2% and 5.8%, respectively. Prevalence of cardiovascular, endocrine, gastrointestinal, and neurologic conditions were higher with the AYA HOPE Index than the other two indices. Forty percent of AYA patients reported service needs, particularly for mental health services (25.2%) and support groups (17.7%). Having ≥2 comorbidities on the AYA index was associated with higher mental health service needs [OR, 2.05; 95% confidence interval (CI), 1.10–3.82] adjusting for demographic and clinical factors. Comorbidities were associated with fair/poor self-reported health status. Conclusion: The AYA HOPE Index is a more comprehensive comorbidity index for AYA cancer patients than existing indices, and the number of comorbidities is associated with service needs and health status. Impact: The AYA HOPE index could identify patients' additional service needs early in therapy. Cancer Epidemiol Biomarkers Prev; 24(12); 1844–9. ©2015 AACR.

Testicular germ cell tumors (TGCT) are ideally suited to agnostic genome-wide association methods because of their known high heritability and homogeneous cell of origin. Here we report the most updated results from the international Testicular Cancer Consortium (TECAC) that extends published findings from our recent meta-analysis, which included 3,558 cases and 13,970 controls from five centers. To these existing samples, we added summary statistics from 300 cases and 151,991 controls provided by deCODE combined with imputed genotypes from an additional 5,602 cases and 5,006 controls assembled from 15 TECAC centers, almost all (97%) of which were centrally genotyped using the Illumina Human Core array. A total of 9,458 TGCT cases were included in our current meta-analysis, representing a 66% increase since our last publication. We identified 14 new and confirmed 41 previously reported loci that surpassed genome-wide significance (5 × 10-8). Fifteen previously reported loci did not reach this threshold, although one did so in subgroup analysis. In addition, 57 new loci attained nominal significance (5 × 10-8 ≤ p ≤ 1 × 10-5). As expected, a substantial proportion of top hits continue to map to genetic regions belonging to biological pathways essential to male germ cell development, sex determination, and chromosomal segregation. For example, newly identified genes include anaphase promoting complex subunit 2 (ANAPC2), BCL2 like 11 (BCL211), anti-Mullerian hormone receptor type 2 (AMHR2), and DEP domain containing mTOR interacting protein (DEPTOR). Our findings also further implicate genes located on the X chromosome, specifically the androgen receptor (AR). In analyses stratified by tumor subtype, we identified GATA-binding protein 4 (GATA4) as a susceptibility locus among men diagnosed with seminoma. Despite a similar effect size observed among men diagnosed solely with non-seminoma (i.e. excluding cases with a mixture of both seminoma and non-seminoma tumors), this marker did not surpass genome-wide significance in this subtype likely because of the smaller case sample size. GATA4 previously has been associated with overall risk of TGCT (Litchfield et al., Nat Genet. 2017). Biological mechanisms for all genome-wide significant loci are currently being explored using publicly available data sets, ATAC-seq of four TGCT cell lines, and SPATIaL-Seq (a novel high-resolution chromatin conformation capture (3C) assay) of the NTERA2 pluripotent human embryonal carcinoma cell line. Additional genotyping of top hits in an independent sample set of 1,103 TGCT cases and 1,210 controls is currently ongoing. Citation Format: Louise C. Pyle, John Pluta, Kevin T. Nead, Nandita Mitra, Javier Benitez, D. Timothy Bishop, Victoria Cortessis, Alberto Ferlin, Jourik Gietema, Mark Greene, Tom Grotmol, Ramneek Gupta, Rob Hamilton, Michelle A. Hildebrandt, Trine B. Haugen, Lambertus Kiemeney, Christian Kubisch, Davor Lessel, Paloma Martin, Thorunn Rafnar, Lorenzo Richiardi, Rolf Skotheim, Clare Turnbull, Fredrik Wiklund, Tongzhang Zheng, Ewa Rajpert-De Meyts, Stephen M. Schwartz, Katherine A. McGlynn, Peter A. Kanetsky, Katherine L. Nathanson, Testicular Cancer Consortium (TECAC). Identification of 14 novel genetic loci for testicular germ cell tumor susceptibility [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2684.

Background: The incidence of oropharyngeal and oral tongue cancers has increased over the last 20 years which parallels increased use of marijuana among individuals born after 1950. Methods: A pooled analysis was conducted comprising individual-level data from nine case–control studies from the United States and Latin America in the INHANCE consortium. Self-reported information on marijuana smoking, demographic, and behavioral factors was obtained from 1,921 oropharyngeal cases, 356 oral tongue cases, and 7,639 controls. Results: Compared with never marijuana smokers, ever marijuana smokers had an elevated risk of oropharyngeal [adjusted OR (aOR), 1.24; 95% confidence interval (CI): 1.06–1.47] and a reduced risk of oral tongue cancer (aOR, 0.47; 95% CI, 0.29, 0.75). The risk of oropharyngeal cancer remained elevated among never tobacco and alcohol users. The risk of oral tongue cancer was reduced among never users of tobacco and alcohol. Sensitivity analysis adjusting for potential confounding by HPV exposure attenuated the association of marijuana use with oropharyngeal cancer (aOR, 0.99; 95% CI, 0.71–1.25), but had no effect on the oral tongue cancer association. Conclusions: These results suggest that the association of marijuana use with head and neck carcinoma may differ by tumor site. Impact: The associations of marijuana use with oropharyngeal and oral tongue cancer are consistent with both possible pro- and anticarcinogenic effects of cannabinoids. Additional work is needed to rule out various sources of bias, including residual confounding by HPV infection and misclassification of marijuana exposure. Cancer Epidemiol Biomarkers Prev; 23(1); 160–71. ©2013 AACR.

Purpose: To determine the differential gene expression between oral squamous cell carcinoma (OSCC) with and without metastasis to cervical lymph nodes and to assess prediction of nodal metastasis by using molecular features. Experimental Design: We used Affymetrix U133 2.0 plus arrays to compare the tumor genome-wide gene expression of 73 node-positive OSCCs with 40 node-negative OSCCs (≥18 months). Multivariate linear regression was used to estimate the association between gene expression and nodal metastasis. Stepwise logistic regression and receiver operating characteristics (ROC) analysis were used to generate predictive models and to compare these with models by using tumor size alone. Results: We identified five genes differentially expressed between node-positive and node-negative OSCCs after adjusting for tumor size and human papillomavirus status: REEP1, RNF145, CTONG2002744, MYO5A, and FBXO32. Stepwise regression identified a four-gene model (MYO5A, RFN145, FBXO32, and CTONG2002744) as the most predictive of nodal metastasis. A leave-one-out ROC analysis revealed that our model had a higher area under the curve (AUC) for identifying occult nodal metastasis compared with that of a model by tumor size alone (respective AUC: 0.85 and 0.61; P = 0.011). A model combining tumor size and gene expression did not further improve the prediction of occult metastasis. Independent validation using 31 metastatic and 13 nonmetastatic cases revealed a significant underexpression of CTONG2002744 (P = 0.0004). Conclusions: These results suggest that our gene expression markers of OSCC metastasis hold promise for improving current clinical practice. Confirmation by others and functional studies of CTONG2002744 is warranted. Clin Cancer Res; 17(8); 2466–73. ©2011 AACR.

Background: Marijuana contains carcinogens similar to tobacco smoke and has been suggested by relatively small studies to increase the risk of head and neck cancer (HNC). Because tobacco is a major risk factor for HNC, large studies with substantial numbers of never tobacco users could help to clarify whether marijuana smoking is independently associated with HNC risk. Methods: We pooled self-reported interview data on marijuana smoking and known HNC risk factors on 4,029 HNC cases and 5,015 controls from five case-control studies within the INHANCE Consortium. Subanalyses were conducted among never tobacco users (493 cases and 1,813 controls) and among individuals who did not consume alcohol or smoke tobacco (237 cases and 887 controls). Results: The risk of HNC was not elevated by ever marijuana smoking [odds ratio (OR), 0.88; 95% confidence intervals (95% CI), 0.67-1.16], and there was no increasing risk associated with increasing frequency, duration, or cumulative consumption of marijuana smoking. An increased risk of HNC associated with marijuana use was not detected among never tobacco users (OR, 0.93; 95% CI, 0.63-1.37; three studies) nor among individuals who did not drink alcohol and smoke tobacco (OR, 1.06; 95% CI, 0.47-2.38; two studies). Conclusion: Our results are consistent with the notion that infrequent marijuana smoking does not confer a risk of these malignancies. Nonetheless, because the prevalence of frequent marijuana smoking was low in most of the contributing studies, we could not rule out a moderately increased risk, particularly among subgroups without exposure to tobacco and alcohol. (Cancer Epidemiol Biomarkers Prev 2009;18(5):1544–51)

Purpose: To determine if gene expression signature of invasive oral squamous cell carcinoma (OSCC) can subclassify OSCC based on survival. Experimental Design: We analyzed the expression of 131 genes in 119 OSCC, 35 normal, and 17 dysplastic mucosa to identify cluster-defined subgroups. Multivariate Cox regression was used to estimate the association between gene expression and survival. By stepwise Cox regression, the top predictive models of OSCC-specific survival were determined and compared by receiver operating characteristic analysis. Results: The 3-year overall mean ± SE survival for a cluster of 45 OSCC patients was 38.7 ± 0.09% compared with 69.1 ± 0.08% for the remaining patients. Multivariate analysis adjusted for age, sex, and stage showed that the 45 OSCC patient cluster had worse overall and OSCC-specific survival (hazard ratio, 3.31; 95% confidence interval, 1.66-6.58 and hazard ratio, 5.43; 95% confidence interval, 2.32-12.73, respectively). Stepwise Cox regression on the 131 probe sets revealed that a model with a term for LAMC2 (laminin γ2) gene expression best identified patients with worst OSCC-specific survival. We fit a Cox model with a term for a principal component analysis-derived risk score marker and two other models that combined stage with either LAMC2 or PCA. The area under the curve for models combining stage with either LAMC2 or PCA was 0.80 or 0.82, respectively, compared with 0.70 for stage alone (P = 0.013 and 0.008, respectively). Conclusions: Gene expression and stage combined predict survival of OSCC patients better than stage alone.

Background: The magnitude of risk conferred by the interaction between tobacco and alcohol use on the risk of head and neck cancers is not clear because studies have used various methods to quantify the excess head and neck cancer burden. Methods: We analyzed individual-level pooled data from 17 European and American case-control studies (11,221 cases and 16,168 controls) participating in the International Head and Neck Cancer Epidemiology consortium. We estimated the multiplicative interaction parameter (ψ) and population attributable risks (PAR). Results: A greater than multiplicative joint effect between ever tobacco and alcohol use was observed for head and neck cancer risk (ψ = 2.15; 95% confidence interval, 1.53-3.04). The PAR for tobacco or alcohol was 72% (95% confidence interval, 61-79%) for head and neck cancer, of which 4% was due to alcohol alone, 33% was due to tobacco alone, and 35% was due to tobacco and alcohol combined. The total PAR differed by subsite (64% for oral cavity cancer, 72% for pharyngeal cancer, 89% for laryngeal cancer), by sex (74% for men, 57% for women), by age (33% for cases 60 years), and by region (84% in Europe, 51% in North America, 83% in Latin America). Conclusions: Our results confirm that the joint effect between tobacco and alcohol use is greater than multiplicative on head and neck cancer risk. However, a substantial proportion of head and neck cancers cannot be attributed to tobacco or alcohol use, particularly for oral cavity cancer and for head and neck cancer among women and among young-onset cases. (Cancer Epidemiol Biomarkers Prev 2009;18(2):541–50)

Oral squamous cell carcinoma (OSCC) is associated with substantial mortality and morbidity. To identify potential biomarkers for the early detection of invasive OSCC, we compared the gene expressions of incident primary OSCC, oral dysplasia, and clinically normal oral tissue from surgical patients without head and neck cancer or preneoplastic oral lesions (controls), using Affymetrix U133 2.0 Plus arrays. We identified 131 differentially expressed probe sets using a training set of 119 OSCC patients and 35 controls. Forward and stepwise logistic regression analyses identified 10 successive combinations of genes which expression differentiated OSCC from controls. The best model included LAMC2, encoding laminin-γ2 chain, and COL4A1, encoding collagen, type IV α1 chain. Subsequent modeling without these two markers showed that COL1A1, encoding collagen, type I α1 chain, and PADI1, encoding peptidyl arginine deiminase, type 1, could also distinguish OSCC from controls. We validated these two models using an internal independent testing set of 48 invasive OSCC and 10 controls and an external testing set of 42 head and neck squamous cell carcinoma cases and 14 controls (GEO GSE6791), with sensitivity and specificity above 95%. These two models were also able to distinguish dysplasia (n = 17) from control (n = 35) tissue. Differential expression of these four genes was confirmed by quantitative reverse transcription-PCR. If confirmed in larger studies, the proposed models may hold promise for monitoring local recurrence at surgical margins and the development of second primary oral cancer in patients with OSCC. (Cancer Epidemiol Biomarkers Prev 2008;17(8):2152–62)

Cigarette smoking is an established cofactor to human papillomavirus (HPV) in the development of cervical and vulvar squamous cell carcinoma (SCC), and may influence risk through an immunosuppressive pathway. Genetic variation in interleukin 2 (IL2), associated in some studies with the inhibition of HPV-targeted immunity, may modify the effect of smoking on the risk of HPV-related anogenital cancers. We conducted a population-based case-only study to measure the departure from a multiplicative joint effect of cigarette smoking and IL2 variation on cervical and vulvar SCC. Genotyping of the four IL2 tagSNPs (rs2069762, rs2069763, rs2069777, and rs2069778) was done in 399 cervical and 486 vulvar SCC cases who had been interviewed regarding their smoking history. Compared with cases carrying the rs2069762 TT genotype, we observed significant departures from multiplicativity for smoking and carriership of the TG or GG genotypes in vulvar SCC risk [interaction odds ratio (IOR), 1.67; 95% confidence interval (CI), 1.16-2.41]. Carriership of one of three diplotypes, together with cigarette smoking, was associated with either a supramultiplicative (TGCT/GGCC; IOR, 2.09; 95% CI, 0.98-4.46) or submultiplicative (TTCC/TGTC; IOR, 0.37; 95% CI, 0.16-0.85 or TGCT/TGCC; IOR, 0.37; 95% CI, 0.15-0.87) joint effect in vulvar cancer risk. For cervical SCC, departure from multiplicativity was observed for smokers homozygous for the rs2069763 variant allele (TT versus GG or GT genotypes; IOR, 1.87; 95% CI, 1.00-3.48), and for carriership of the TTCC/TTCC diplotype (IOR, 2.08; 95% CI, 1.01-4.30). These results suggest that cervical and vulvar SCC risk among cigarette smokers is modified by genetic variation in IL2. (Cancer Epidemiol Biomarkers Prev 2008;17(7):1790–9)

Variation in human major histocompatibility genes may influence the risk of squamous cell cervical cancer (SCC) by altering the efficiency of the T-cell–mediated immune response to human papillomavirus (HPV) antigens. We used high-resolution methods to genotype human leukocyte antigen (HLA) class I (A, B, and Cw) and class II (DRB1 and DQB1) loci in 544 women with SCC and 542 controls. Recognizing that HLA molecules are codominantly expressed, we focused on co-occurring alleles. Among 137 allele combinations present at >5% in the case or control groups, 36 were significantly associated with SCC risk. All but one of the 30 combinations that increased risk included DQB1*0301, and 23 included subsets of A*0201-B*4402-Cw*0501-DRB1*0401-DQB1*0301. Another combination, B*4402-DRB1*1101-DQB1*0301, conferred a strong risk of SCC (odds ratio, 10.0; 95% confidence interval, 3.0–33.3). Among the six combinations that conferred a decreased risk of SCC, four included Cw*0701 or DQB1*02. Most multilocus results were similar for SCC that contained HPV16; a notable exception was A*0101-B*0801-Cw*0701-DRB1*0301-DQB1*0201 and its subsets, which were associated with HPV16-positive SCC (odds ratio, 0.5; 95% confidence interval, 0.3–0.9). The main multilocus associations were replicated in studies of cervical adenocarcinoma and vulvar cancer. These data confirm that T helper and cytotoxic T-cell responses are both important cofactors with HPV in cervical cancer etiology and indicate that co-occurring HLA alleles across loci seem to be more important than individual alleles. Thus, certain co-occurring alleles may be markers of disease risk that have clinical value as biomarkers for targeted screening or development of new therapies. [Cancer Res 2008;68(9):3532–9]

The incidence of testicular germ cell carcinoma (TGCC) is highest among men ages 20 to 44 years. Exposure to relatively high circulating maternal estrogen levels during pregnancy has long been suspected as being a risk factor for TGCC. Catechol (hydroxylated) estrogens have carcinogenic potential, thought to arise from reactive catechol intermediates with enhanced capability of forming mutation-inducing DNA adducts. Polymorphisms in maternal or offspring genes encoding estrogen-metabolizing enzymes may influence prenatal catechol estrogen levels and could therefore be biomarkers of TGCC risk. We conducted a population-based, case-parent triad study to evaluate TGCC risk in relation to maternal and/or offspring polymorphisms in CYP1A2, CYP1B1, CYP3A4, and CYP3A5. We identified 18- to 44-year-old men diagnosed with invasive TGCC from 1999 to 2004 through a population-based cancer registry in Washington State and recruited cases and their parents (110 case-parent triads, 50 case-parent dyads). Maternal or offspring carriage of CYP1A2 −163A was associated with reduced risk of TGCC [maternal heterozygote relative risk (RR), 0.6; 95% confidence interval (95% CI), 0.2-1.7; offspring heterozygote RR, 0.7; 95% CI, 0.3-1.5)]. Maternal CYP1B1 48Gly homozygosity was associated with a 2.7-fold increased risk of TGCC (95% CI, 0.9-7.9), with little evidence that Leu432Val or Asn453Ser genotypes were related to risk. Men were also at increased risk of TGCC if they carried the CYP3A4 −392G (RR, 7.0; 95% CI, 1.6-31) or CYP3A5 6986G (RR, 2.4; 95% CI, 1.1-5.6) alleles. These results support the hypothesis that maternal and/or offspring catechol estrogen activity may influence sons' risk of TGCC.

Tobacco and alcohol consumption are the major risk factors for head and neck cancer, likely due to DNA-damaging processes. Genetic variations in DNA repair genes may affect an individual's susceptibility to head and neck cancer. Pooling data and DNA specimens from three case-control studies in western Washington State, North Carolina, and Puerto Rico, totaling 555 cases (430 whites) and 792 controls (695 whites), we studied the risk of head and neck cancer in relation to common nonsynonymous single-nucleotide polymorphisms in four DNA repair genes: MGMT (Leu84Phe and Ile143Val), XRCC1 (Arg399Gln), XPD (Lys751Gln), and XRCC3 (Thr241Met). All single-nucleotide polymorphisms were assayed in a single laboratory. Among whites, carriage of the MGMT Phe84 [odds ratio (OR), 0.71; 95% confidence interval (95% CI), 0.51-0.98] or Val143 (OR, 0.66; 95% CI, 0.47-0.92) allele was associated with a decreased risk of head and neck cancer; the haplotype distribution for MGMT differed significantly between cases and controls (covariate-adjusted global permutation test, P = 0.012). The XRCC1 GlnGln399 genotype was also associated with decreased risk among whites (OR, 0.56; 95% CI, 0.32-0.94), whereas XPD751 and XRCC3241 were not associated with risk. Alcohol-related risks tended to vary with DNA repair genotypes, especially for MGMT variants, whereas no effect modification was noted with tobacco use. Consistent findings from three case-control studies suggest that selected DNA repair enzymes may play a role in head and neck carcinogenesis.

2-Hydroxylated metabolites of estrogen have been shown to have antiangiogenic effects and inhibit tumor cell proliferation, whereas 4-hydroxylated metabolites have been implicated in carcinogenesis. We examined whether polymorphisms in certain genes involved in estrogen metabolism are associated with endometrial cancer risk in a population-based case-control study with 371 cases and 420 controls. Based on previously published genotype-phenotype correlation studies, we defined variant alleles thought to increase estrogen 2-hydroxylation as presumptively low-risk (CYP1A1 m1 T6235C and m2 Ile462Val) and those thought to increase estrogen 4-hydroxylation as high-risk (CYP1A1 m4 Thr461Asn, CYP1A2 A734C, and CYP1B1 Leu432Val). Odds ratios (OR) and 95% confidence intervals (95% CI) were calculated using unconditional logistic regression. Carrying at least one CYP1A1 m1 or m2 variant allele was associated with a decreased risk of endometrial cancer [ORs (95% CIs), 0.64 (0.44-0.93) and 0.54 (0.30-0.99), respectively]. No strong alteration in risk was observed among women with any of the putative high-risk alleles. When CYP1A1, CYP1A2, and CYP1B1 genotypes were combined and ranked by the number of putative low-risk genotypes carried, women with four or five low-risk genotypes had a reduced risk of endometrial cancer (OR, 0.29; 95% CI, 0.15-0.56) compared with women with one or none. No appreciable alteration in risk was observed among women carrying two or three low-risk genotypes. Some of our findings are consistent with the hypothesis that increased estrogen 2-hydroxylation is associated with decreased endometrial cancer risk, but replication of these results is required before any firm conclusions can be reached.

Circulating hormones are associated with mammographic density, an intermediate marker of breast cancer risk. Differences in circulating hormones, including estrone and testosterone, have been observed in premenopausal women based on their capacity to metabolize daidzein, an isoflavone found predominantly in soybeans. Equol and O-desmethylangolensin (O-DMA) are products of intestinal bacterial metabolism of daidzein. There is interindividual variability in the capacity to produce daidzein metabolites; individuals can be equol producers or non-producers and O-DMA producers or non-producers. We tested the hypothesis that daidzein-metabolizing phenotypes are associated with mammographic density. Participants were recruited from among 92 sedentary, postmenopausal women, ages 50 to 75 years, who participated in a 1-year physical activity intervention. Pre-intervention mammographic density was determined using a computer-assisted, gray-scale thresholding technique. Fifty-five of these women consumed supplemental soy protein (>10 mg daidzein/d) for 3 days and collected a first-void urine sample on the fourth day to determine daidzein-metabolizing phenotypes. Equol and O-DMA concentrations were measured using gas chromatography-mass spectrometry. Associations between daidzein-metabolizing phenotypes and percent mammographic density were adjusted for age, maximum adult weight, gravidity, family history of breast cancer, and serum follicle-stimulating hormone and free testosterone concentrations. Mammographic density was 39% lower in equol producers compared with non-producers (P = 0.04). O-DMA producers had mammographic density 69% greater than non-producers (P = 0.05). These results suggest that particular intestinal bacterial profiles are associated with postmenopausal mammographic density, and these associations are not entirely explained by differences in reproductive or anthropometric characteristics or circulating hormones.

Previous laboratory investigations, case reports, and a hospital-based case-control study have suggested that marijuana use may be a risk factor for squamous cell head and neck cancer. We conducted a population-based case-control study to determine whether marijuana use is associated with the development of oral squamous cell carcinoma (OSCC). Case subjects (n = 407) were 18–65-year-old residents of three counties in western Washington State who were newly diagnosed with OSCC from 1985 through 1995. Control subjects (n = 615), who were similar to the cases with respect to age and sex, were selected from the general population using random-digit telephone dialing. Lifetime histories of marijuana use and exposure to known OSCC risk factors were ascertained using a structured questionnaire. Information on genetic polymorphisms in glutathione S-transferase enzymes was obtained from assays on participant DNA. Odds ratios for associations with features of marijuana use were adjusted for sex, education, birth year, alcohol consumption, and cigarette smoking. A similar proportion of case subjects (25.6%) and control subjects (24.4%) reported ever use of marijuana (adjusted odds ratio, 0.9; 95% confidence interval, 0.6–1.3). There were no trends in risk observed with increasing duration or average frequency of use or time since first or last use. No subgroup defined by known or suspected OSCC risk factors (age, cigarette smoking, alcohol consumption, and genetic polymorphisms) showed an increased risk. Marijuana use was not associated with OSCC risk in this large, population-based study.

Genome-wide association studies (GWAS) have already identified 21 genetic loci associated with testicular germ cell tumor (TGCT). Many of the loci contain biologically plausible genes that function in male germ cell maturation and differentiation, as well as KIT-MAPK signaling and chromosomal segregation. We recently formed the international TEsticular CAncer Consortium (TECAC), and pooled data from the three published and one unpublished GWAS (3,556 TGCT cases and 13,969 controls) to identify additional novel susceptibility loci. We imputed across the data set using the 1,000 Genomes Project version 3 and conducted a fixed effects meta-analysis, including the first analysis of the X chromosome. Eleven new loci achieved genome-wide significance level (p Citation Format: Zhaoming Wang, Peter A. Kanetsky, Katherine A. McGlynn, D. Timothy Bishop, Charles C. Chung, Marlene D. Dalgaard, Tom Grotmol, Mark H. Greene, Ramneek Gupta, Trine B. Haugen, Kevin Litchfield, Jennifer T. Loud, Nandita Mitra, Kasper Nielsen, Clare Turnbull, Ewa Rajpert-DeMeyts, Saran Vardhanabhuti, Fredrik Wiklund, Stephen Schwartz, Stephen J. Chanock, Katherine L. Nathanson, TECAC consortium. Imputation and meta-analysis of five genome-wide association studies identify multiple new loci associated with testicular germ cell tumor. [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 843. doi:10.1158/1538-7445.AM2015-843

Background: Numerous studies have assessed associations between lipid levels and risk of colorectal neoplasia, but findings have been inconsistent. Recent genetic studies have identified >100 loci associated with lipid levels. Knowledge of the magnitude and direction of genetic effects permits evaluation of allele scores that can serve as proxies for phenotypes. Methods: We collected information on blood lipids as part of a colonoscopy study among enrollees, ages 25-79, of Group Health, a large healthcare system in Washington State. Low-density lipoprotein (LDL), high-density lipoprotein (HDL), triglycerides (TG), and total cholesterol (TC) measurements were extracted from electronic medical records for 98% of participants. For each participant, we identified the highest LDL, TG, and TC measurement (zenith) and lowest HDL measurement (nadir) prior to colonoscopy. All participants were genotyped for 96 single-nucleotide polymorphisms (SNPs) identified by the Global Lipids Genetics Consortium. For each lipid trait, we estimated 3 associations: 1) phenotype-polyp odds ratios (OR) with 95% confidence intervals (CI) comparing non-advanced adenoma cases to controls, advanced adenoma cases (≥10 mm in diameter, with villous components, or high-grade dysplasia) to controls, and non-adenomatous polyp cases to controls from polytomous logistic regression; 2) genotype-phenotype associations from linear regression; and 3) genotype-polyp ORs from two-stage linear-logistic regression. Results: In total, 1,791 participants had information on phenotype and genotype (518 non-advanced adenoma cases, 139 advanced adenoma cases, 380 non-adenomatous polyp cases, and 754 polyp-free controls). Individuals with advanced adenomas were more likely than controls to have higher LDL and TG (adjusted OR per 20 mg/dL increase in zenith LDL: 1.16, CI 1.03-1.30 and OR per 40 mg/dL increase in zenith TG: 1.09, 1.03-1.16). Associations from allele scores were in the same direction (OR per increase in allele score scaled to be comparable to a 20 mg/dL LDL increase: 1.17, CI 0.78-1.75, and OR per increase in allele score scaled to be comparable to a 40 mg/dL TG increase: 1.12, 0.91-1.38). SNPs most strongly related to phenotype, however, were not associated with polyps, and SNPs that were, including variants of NAT2, MC4R, and APOE, may function through alternative pathways. Analyses among statin-naïve participants were not meaningfully different. Conclusions: Mendelian randomization estimates did not achieve statistical significance, but the direction of associations suggests that polyps are more prevalent among those with inherited susceptibility to increased LDL and TG. Results also highlight difficulties interpreting results from Mendelian randomization analyses of traits with complex biology, given that variants may be pleiotropic and not suitable for use as instrumental variables. Citation Format: Michael N. Passarelli, Polly A. Newcomb, Karen W. Makar, Andrea N. Burnett-Hartman, John D. Potter, Melissa P. Upton, Lee-Ching Zhu, Michael E. Rosenfeld, Stephen M. Schwartz, Carolyn M. Rutter. Blood lipids, colorectal adenomas, and non-adenomatous polyps: A comparison of associations from clinical measurements and Mendelian randomization. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 294. doi:10.1158/1538-7445.AM2014-294

Oral and oropharyngeal squamous cell carcinomas (OSCC) are among the most common cancers worldwide. The overall 5-yr survival rate of OSCC patients is approximately 50% but is higher than 80% in early-stage patients. The high mortality of OSCC is due in part to late diagnosis and high recurrence rates. Thus, early detection of OSCC and identification of a good predictor of disease progression are important in improving OSCC outcomes. To identify potential markers for early diagnosis and disease progression, we used Affymetrix U133 plus 2.0 arrays to examine the gene expression profiles of 167 primary OSCC from newly diagnosed patients, 58 “normal” oral mucosa from oral cancer patients (NCA) and 45 normal oral mucosa from patients without cancer (controls), all of whom were enrolled at three University of Washington-affiliated medical centers from 2003 to 2008. We first compared gene expression in the 45 controls to the 167 tumors using ANOVA implemented in Partek® Genomics Suite™ software, adjusting for age, sex, cigarette-smoking, alcohol use and HPV status. We found 2,596 probe sets differentially expressed between tumors and controls, using the criteria of a FDR of 0.05 and at least a two-fold difference in the expression level. We then used linear regression to compare the gene expression level of these 2,596 probe sets between controls and NCA, and between NCA and tumors. Among 2,596 probe sets, 60 were significantly up-regulated and 11 were significantly down-regulated in both comparisons using the Bonferroni correction with a p-value of 1.93x10-5 and a difference in gene expression greater than 1 standard deviation of the expression in NCA. A cluster analysis on gene expression of the 71 probe sets identified a subgroup of 45 OSCC patients with highest/lowest expression of these probe sets. These 45 OSCC patients had a higher risk of disease progression after treatment (defined by persistence or recurrence of OSCC) and higher OSCC-specific mortality when compared with the remaining 122 OSCC patients; the respective Cox regression hazard ratio (HR) and 95% CI, adjusted for age, sex, and stage for disease progression and OSCC-specific mortality were 2.7 (1.5-4.6) and 3.8 (2.1-7.0). The top gene associated with both outcomes was dermatan sulfate epimerase (DSE). A high level of DSE expression in tumor (higher than mean expression among tumors) was associated with a higher risk of disease progression (HR 2.8, 95% CI: 1.5-5.1) and of OSCC-specific mortality (HR 6.0, 95% CI: 2.9-12.2) after adjusting for age, sex and stage. DSE encodes a tumor-rejection antigen that could potentially serve as a target for cancer immunotherapy. In conclusion, we identified 71 probe sets in which dysregulation occurred in both NCA and cancer samples. Some of these genes could potentially serve as early diagnostic markers, markers to predict disease progression, or as potential targets for new therapy. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 724. doi:1538-7445.AM2012-724

History of any type of allergy is associated with a decreased risk of multiple types of cancers, including colorectal, cervical, esophageal, pancreatic, stomach, leukemia and glioma. It is proposed that having a history of allergies may prevent cancer by enhancing immune surveillance against tumor cells early in disease development and/or against carcinogenic infectious agents. While a few prior studies suggest an inverse relationship between history of allergies and oral squamous cell carcinoma (OSCC); they did not distinguish between oropharyngeal cancer and oral cavity cancers, and did not account for HPV infection. We compared 400 OSCC cases newly diagnosed between 1985-1995 with 613 general population controls from the Seattle-Puget Sound metropolitan areas with respect to self-reported history of allergy to each of the following: dust, pollen, mold, dogs, cats, bee stings, drugs and food. We estimated odds ratios (OR) and 95% confidence intervals (CI), adjusted for age, education level, sex, race, pack years of cigarette smoking, and lifetime average alcoholic drinks per week. We observed a small inverse association between history of any allergy and OSCC (OR=0.81, 95% CI, 0.61-1.08). This inverse association was present only for dust (OR=0.70; 95% CI,0.43-1.13), pollen (OR=0.72; 95% CI,0.51-1.02) and mold (OR=0.34; 95% CI,0.14-0.83). Additional adjustment for HPV-16 seropositivity did not materially affect the estimates. In exploratory analyses, we examined associations with dust, pollen or mold more closely, and observed stronger associations with oropharyngeal cancer (OR=0.56; 95% CI, 0.35-0.90) than with oral cavity cancer (OR=0.71; 95% CI, 0.49-1.05); although a polytomous regression model did not suggest heterogeneity by site (p=0.5). There was no effect modification by sex, cigarette smoking, alcohol consumption, HPV-16 seropositivity (for all sites or oropharynx alone), or herpes simplex virus 1 seropositivity. The inverse association with dust/pollen/mold was near null for 30 cases of oncogenic HPV DNA-positive oropharyngeal cancer (OR=0.86; 95% CI, 0.38-1.94) and stronger for 46 cases of oncogenic HPV DNA-negative oropharyngeal cancer (OR=0.53; 95% CI, 0.24-1.19). These results suggest that a history of allergies to dust, pollen or mold may confer some protection against OSCC. That the inverse association was particularly apparent for oropharyngeal SCC would be consistent with heightened immune surveillance, due to the substantial amount of lymphatic tissue in this organ. Whether the inverse association involves an immune response to HPV or other antigen, or other carcinogenic mechanism, remains to be determined in more definitive studies. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5491. doi:1538-7445.AM2012-5491

Proceedings: AACR 103rd Annual Meeting 2012‐‐ Mar 31‐Apr 4, 2012; Chicago, IL Testicular germ cell tumors (TGCT) are the most common cancers in young men, and several lines of evidence point to their substantial genetic component. To date, genome-wide association (GWA) studies have uncovered six TGCT susceptibility loci. Together these loci account for 11-16% of TGCT disease risk to first degree relatives, arguing that additional genetic variation contributing to TGCT susceptibility must exist. Recently we and others indentified variation at DMRT1 associated with TGCT risk. DMRT1 is highly conserved across many species and plays a crucial role in male gonadal development and sex determination. Deletions in DMRT1 lead to male to female (XY) sex reversal. Therefore, we undertook a targeted pathway-based analysis of genes influencing XY sex reversal to identify associations with TGCT risk. Through review of the literature, we identified 27 additional genes in which mutations or deletions are associated with XY sex reversal. Using our existing GWA data in 349 cases and 919 controls, ∼570 risk markers in the 28 candidate loci were evaluated for replication based on a significance threshold of p < 2 x 10−5 for at least two SNP markers. Two markers that mapped near to MAP3K1 and FGF9 meet these criteria. We genotyped the most statistically significant SNP markers in the MAP3K1 ([rs252906][1]; p = 1.89 x 10Δ5) and FGF9 ([rs628137][2]; p = 1.66 x 10Δ5) region in two independent replication sample sets, consisting of 439 cases of TGCT and 960 controls and of 768 cases and 674 controls respectively. In combined analysis, TGCT cases had elevated odds of carriage of the [rs252906][1] major T allele [per-allele odds ratio (OR) = 1.24, 95% confidence interval (CI) 1.16, 1.377; P = 5.87 × 10−5] and the [rs628137][2] major G allele (per allele OR = 1.20, 95% CI 1.08, 1.34, P = 5.28 x 10−4 compared with controls. Similarly to KITLG, the susceptibility locus most strongly associated with TGCT risk, FGF9 is associated with proliferation, migration and survival of primordial germ cells, the progenitor cells for human TGCT. MAP3K1 and FGF9 play a crucial role in normal testes development. These finding strengthen the supposition that variation in genes influencing sex determination, and male germ cell development and differentiation play an important role in inherited TGCT susceptibility. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 2622. doi:1538-7445.AM2012-2622 [1]: /lookup/external-ref?link_type=GEN&access_num=rs252906&atom=%2Fcanres%2F72%2F8_Supplement%2F2622.atom [2]: /lookup/external-ref?link_type=GEN&access_num=rs628137&atom=%2Fcanres%2F72%2F8_Supplement%2F2622.atom

Proceedings: AACR 102nd Annual Meeting 2011‐‐ Apr 2‐6, 2011; Orlando, FL The matrix metalloproteinase genes have frequently been identified as important in cancer development and progression, including in oral squamous cell carcinoma (OSCC). These proteases are believed to cause degradation of the extracellular matrix and basement membranes and therefore to play a key role in tumor metastasis, cell migration, cancer cell growth, and angiogenesis. We compared MMP1 expression levels in primary OSCC (n=119) with expression levels in normal oral epithelium from control subjects (n=35), as measured by Affymetrix U133 2.0 Plus GeneChip arrays, adjusting for age and sex. MMP1 was expressed more than 200-fold higher in OSCC compared with normal control epithelium. qRT-PCR results on 30 of the cases and 22 of the controls confirmed the substantial differential expression of MMP1, with a difference in mean threshold cycle (Ct) value for cases versus controls of 9.85 (95% CI, 8.74-10.97, p

The presence of lymph node metastasis is associated with a 50% reduction of 5-year survival in patients with oral squamous cell carcinoma (OSCC). We set out to combine DNA copy number aberrations (CNAs) with gene expression profiles, to identify CNAs-associated genes dysregulated in metastatic OSCC, and determine whether these genes can be targeted to selectively kill metastatic OSCC tumor cells. Toward this end, we interrogated DNA and RNA of the same OSCC cell populations laser micro-dissected from non-metastatic primary tumors (n=17) and metastatic lymph nodes (n=20) using Affymetrix 250K Nsp single-nucleotide polymorphism (SNP) arrays and U133 Plus 2.0 arrays, respectively. With a false discovery rate (FDR) < 5%, 1988 transcripts were found to be differentially expressed between primary and metastatic OSCC. Out of these, 114 transcripts showed significant correlation between their DNA copy number alternations (estimated using SNPs within 250 kb upstream and downstream of the transcript) and gene expression (FDR < 0.01). Among these CNA-correlated transcripts, 95 had significantly different DNA copy numbers between metastatic and non-metastatic OSCC (FDR Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4983. doi:10.1158/1538-7445.AM2011-4983

Background. Head and neck cancer (HNC) is a significant cause of morbidity and mortality, with over half a million cases worldwide each year. While high fruit and vegetable intake is thought to be protective against HNC, it is unclear whether vitamin and mineral supplement intake is associated with decreased risk of these malignancies. Previous case-control studies in the US have suggested vitamin E supplement intake is inversely associated with oral cavity and pharyngeal cancers. In a large randomized controlled trial, supplementation with alpha-tocopheryl acetate and/or beta-carotene was not associated with upper aerodigestive tract cancer incidence; however an inverse association was suggested for early stage laryngeal cancers. Methods. We analyzed individual-level pooled data from 12 case-control studies (7,002 HNC cases and 8,383 controls) participating in the International Head and Neck Cancer Epidemiology (INHANCE) consortium. There were a total of 2,028 oral cavity cancer cases, 2,465 pharyngeal cancer cases, and 874 laryngeal cancer cases. Odds ratios (OR) and 95% confidence intervals (CIs) for ever use of any vitamins, multivitamins, vitamin A, vitamin C, vitamin E, and calcium, beta carotene, iron, selenium, and zinc supplement were assessed. We further examined frequency, duration and cumulative exposure of each vitamin or mineral when possible. All ORs were adjusted for age, sex, race/ethnicity, study center, education level, pack-years of smoking, frequency of alcohol drinking and fruit/vegetable intake. Results. A decreased risk of HNC was observed with ever use of vitamin C (OR=0.76, 95%CI=0.59-0.96) and with ever use of calcium supplements (OR=0.64, 95%CI=0.42-0.97). There did not appear to be any associations between HNC risk and ever use of any vitamins (OR=0.87, 95%CI=0.71-1.06), multiple vitamins (OR=0.99, 95%CI=0.82-1.19), vitamin A (OR=1.04, 95%CI=0.54-1.98), vitamin E (OR=0.71, 95%CI=0.45-1.11), beta-carotene (OR=1.35, 95%CI=0.65-2.81), iron (OR=0.79, 95%CI=0.54-1.16), selenium (OR=1.21, 95%CI=0.35-4.22) or zinc (OR=0.88, 95%CI=0.34-2.26). Dose-response trends were suggested for years of use of vitamin C (p for trend=0.03) and calcium (p 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 2818.

To examine the impact of copy number aberrations (CNA) on gene expression in oral squamous cell carcinoma (OSCC) metastasis, we used Affymetrix 250K Nsp single nucleotide polymorphism arrays and U133 Plus 2.0 expression arrays, respectively, to interrogate the DNA and RNA from metastatic OSCC cells isolated using laser capture microdissection from lymph nodes of 20 patients. Overall, CNA accounted for expression changes in about 30% of the transcripts studied. With a false discovery rate < 1%, 530 transcripts (461 genes) were identified as to have significantly correlated CNA and expression. When tested using an independent dataset with expression profiles for 148 primary OSCC and 45 normal oral mucosa, the genes were enriched with candidates associated with OSCC (vs. normal oral mucosa) (n=174 transcripts differentially expressed) and OSCC-specific survival (n=32 transcripts associated), as compared to a random gene set of the same size (p < 0.001). We then fit two Cox models using the first two principal components derived from these two gene sets (‘174-transcript PCA’ or ‘32-transcript PCA’), and two other models that combined stage with either the 174- or the 32-transcript PCA score. The Area Under the Curve (AUC) for a model combining stage with 174-gene PCA was significantly higher than that for a model with stage alone (AUC=0.80 vs. 0.72, p = 0.042). Further investigation is warranted to confirm these findings and to examine the biologic role of these copy number-associated transcripts in OSCC metastasis and their potential as therapeutic 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 2209.

MicroRNAs (miRNAs), a group of small non-coding RNAs, regulate post transcriptional gene expression by binding to their target messenger RNAs resulting in their degradation or reduced translation. Evidence suggests that miRNAs are involved in the pathogenesis of cancer and could potentially be used as diagnostic or prognostic markers. MicroRNAs have been studied extensively in solid tumors such as breast, lung and colon cancers; however, there are few studies of miRNA in oral squamous cell carcinoma (OSCC). The purpose of this study was to investigate the potential role of miRNA in OSCC. We selected tumors from 40 patients with OSCC (10 HPV-positive oral cavity tumors, 10 HPV-negative oral cavity tumors, 10 HPV-positive oropharyngeal tumors, and 10 HPV-negative oropharyngeal tumors) and normal oral tissues from 10 patients without cancer (all HPV-negative) who were treated at three University of Washington affiliated medical centers between December 2003 and April 2007 and were enrolled in the study with informed consent. The tissues were obtained at the time of surgery and DNA and RNA were simultaneously extracted using TRIzol method (Invitrogen, Carlsbad, CA). RNA was tested using miRCURY LNATM microRNA v.11.0 (Exiqon, Vedbaek, Denmark). This array covers 1,282 mature human miRNAs and 80 viral miRNAs. Using TIGR software and significance analysis of microarrays (SAM) and a cut point of 2-fold change, we found 30 miRNAs (12 up-regulated and 18 down-regulated) differentially expressed between OSCC and controls. We found no difference in miRNA expression between HPV-positive and HPV-negative tumors, both overall and among oropharyngeal tumors. To confirm the Exiqon array results, we conducted qRT-PCR of miRNAs that had >4-fold change (miR-223: 6.2-fold; miR-21: 4.5-fold); the correlation between miRNA expression as measured by Exiqon array and qRT-PCR was 0.63 (p 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 3052.

B127 The incidence of testicular germ cell carcinoma (TGCC), the most common malignancy among men aged 20-34, has increased several-fold during the past several decades. The etiology of TGCC is poorly understood and the cause of the increased incidence is unknown. Established risk factors include age, race, personal history of undescended testes, and family history of TGCC, none of which is modifiable. Alcohol is an established testicular toxicant, causing testicular atrophy, gynecomastia, and disrupted sexual function in alcoholic men. In both alcoholics and non-alcoholic men with moderate intake, alcohol depresses levels of circulating testosterone and luteinizing hormone. We hypothesized that alcohol consumption, particularly during adolescence, increases the risk of TGCC. A single previous epidemiologic analysis of alcohol intake and TGCC found no association. We conducted a population-based case-control study among 18-44 year-old male residents of three Washington State counties. Cases (n=400) were diagnosed during 1999-2006 with a first, primary TGCC. Controls (n=1,020) were men of similar age with no history of TGCC from the same population identified through random-digit telephone dialing. Questionnaires elicited information on demographic, medical, and lifestyle factors, including usual consumption of beer, wine, and liquor during the 5 years prior to reference date and during grades 7-12. We used logistic regression models to compute odds ratio (OR) estimates and 95% confidence intervals (CI) for TGCC in men that consumed 1-6, 7-13, and 14+ alcoholic beverages per week compared to those that consumed none, adjusting for age, education, smoking, and current marijuana use. A higher proportion of cases than controls (73% vs. 66%) reported drinking 1 or more alcoholic beverages per week during the five years prior to reference date, and during grades 9-12 (44% vs. 37%). Consumption of 14+ alcoholic drinks/week was associated with an approximately 60% increased risk of TGCC (OR=1.62; 95% CI: 1.02-2.58) for consumption during the 5 years prior to reference date, and for consumption during grades 9-12 (OR=1.56; 95% CI: 0.99-2.47). The risk of TGCC appeared to be similar for consumption of beer, wine, or liquor, and for consumption of alcoholic beverages during adolescence versus consumption during adulthood. The risk of TGCC associated with 14+ alcoholic drinks/week was higher among men diagnosed with nonseminomas (OR=2.28; 95% CI: 1.08-4.82) than among men diagnosed with seminomas (OR=1.40 95%CI: 0.80-2.42). Our results indicate that consumption of 14 or more alcoholic beverages per week during adolescence or adulthood may be associated with a doubling of the risk of non-seminomatous TGCC. This is the first study to report an association between alcohol consumption and an increased risk of TGCC. Additional studies are needed to clarify the relationship between alcohol intake and TGCC. Citation Information: Cancer Prev Res 2008;1(7 Suppl):B127.