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2971.Full-Text.Pdf Published OnlineFirst March 28, 2014; DOI: 10.1158/1078-0432.CCR-13-2567 Clinical Cancer Imaging, Diagnosis, Prognosis Research Steroidogenic Germline Polymorphism Predictors of Prostate Cancer Progression in the Estradiol Pathway Eric Levesque 1,2, Isabelle Laverdiere 1, Etienne Audet-Walsh1, Patrick Caron1,Melanie Rouleau1, Yves Fradet2, Louis Lacombe2, and Chantal Guillemette1 Abstract Purpose: Reliable biomarkers that predict prostate cancer outcomes are urgently needed to improve and personalize treatment approaches. With this goal in mind, we individually and collectively appraised common genetic polymorphisms related to estradiol metabolic pathways to find prostate cancer prognostic markers. Methods: The genetic profiles of 526 men with organ-confined prostate cancer were examined to find common genetic polymorphisms related to estradiol metabolic pathways and these findings were replicated in a cohort of 213 men with more advanced disease (follow-up time for both cohorts, >7.4 years). Specifically, we examined 71 single-nucleotide polymorphisms (SNP) in SULT2A1, SULT2B1, CYP1B1, COMT, CYP3A4, CYP3A5, CYP3A43, NQO1, and NQO2 and assessed the impact of the SNPs alone and in combination on prostate cancer progression and on circulating hormone levels. Results: According to a multivariate analysis, CYP1B1 (rs1800440), COMT (rs16982844), and SULT2B1 (rs12460535, rs2665582, rs10426628) were significantly associated with prostate cancer progression and hormone levels. Remarkably, by combining the SNP information with previously identified HSD17B2 markers, the patients could be stratified into four distinct prognostic subgroups. The most prominent association was observed for the eight-marker combination [CYP1B1 (rs1800440), SULT2B1 (rs12460535, rs2665582, and rs10426628), and HSD17B2 (rs4243229, rs1364287, rs2955162, and rs1119933)]. Conclusion: This study identified specific germline variations in estradiol metabolism–related pathways, namely CYP1B1, SULT2B1,andHSD17B2, as novel prognostic markers that are cumulatively associated with increased risk of prostate cancer progression. This panel of markers warrants additional investigation and validation to help stratify patients according to their risk of progression. Clin Cancer Res; 20(11); 2971–83. Ó2014 AACR. Introduction Simple, noninvasive, and reliable molecular markers are Prostate cancer is the most common cancer in men and necessary to identify men at high risk for prostate cancer the second leading cause of cancer death in North America progression. Such prognostic markers for use in localized, (1). A clinically relevant, prognostic molecular signature locally advanced, and metastatic settings would consider- defining aggressive and indolent prostate cancer is eagerly ably improve prostate cancer management. awaited to personalize treatment approaches. Indeed, pros- Recently, circulating tumor cells and whole-blood gene- tate cancer is a complex genetic disease, and its clinical expression signatures were elegantly shown to be potential heterogeneity underscores the need to identify biomarkers biomarkers of cancer progression, and use of these markers for prognostication purposes at all stages of prostate cancer. enabled identification of several differentially regulated genes in immune or androgen signaling that may be useful for predicting outcomes for patients with castration-resis- Authors' Affiliations: 1Pharmacogenomics Laboratory, Centre Hospitalier tant prostate cancer (2, 3). Another useful and much sim- Universitaire de Quebec (CHU de Quebec) Research Center and Faculty of pler approach to predict prostate cancer outcomes relies on Pharmacy; and 2CHU de Quebec Research Center and Faculty of Medicine, Laval University, Quebec, Canada identifying key inherited genetic variations involved in disease progression (4, 5). In fact, potential molecular Note: Supplementary data for this article are available at Clinical Cancer Research Online (http://clincancerres.aacrjournals.org/). determinants of clinical outcomes have been identified as components encoded by immune system genes (6, 7), sex Corresponding Author: Chantal Guillemette, CHU de Quebec Research Center, Laval University, 2705 Boul Laurier, R4720 Quebec G1V 4G2, steroid–related genes (4, 8), and, with conflicting results, Canada. Phone: 418-654-2296; Fax: 418-654-2761; E-mail: prostate cancer risk alleles (9–12). The importance of [email protected] studying host steroidogenic pathways comes from observa- doi: 10.1158/1078-0432.CCR-13-2567 tions that prostate cells possess the enzymatic machinery for Ó2014 American Association for Cancer Research. intracrine conversion of precursors into more potent www.aacrjournals.org 2971 Downloaded from clincancerres.aacrjournals.org on September 27, 2021. © 2014 American Association for Cancer Research. Published OnlineFirst March 28, 2014; DOI: 10.1158/1078-0432.CCR-13-2567 Levesque et al. to 90% to 100% (20). Moreover, knockout of CYP19A1 Translational Relevance (encoding aromatase) in mice, which prevents estrogen Although there has been a decline in mortality rate due production, elevated the circulating T levels of the mice and to early detection and better therapies, our ability to caused enlargement of their prostates, but this knockout did predict prostate cancer progression and metastatic not cause the mice to develop prostate cancer (21, 22). These behavior of a patient’s cancer is still very limited. This observations are consistent with the conclusion that both variability in clinical outcome underscores the need to androgen and estrogen are required for the development of identify physiopathologic changes, in tumor cells and its prostate cancer. Furthermore, Cussenot and colleagues dem- microenvironment, associated with disease progression. onstrated the role of key estrogen metabolism–related genes, Despite the fact that estrogens have been overshadowed e.g., CYP1B1 and catechol-O-methyltransferase (COMT),ina by androgens in the last decades, several pieces of evi- large and homogeneous French cohort with prostate cancer dence suggested that estrogens cooperate with andro- carcinogenesis (23). Given these important findings sup- gens to perpetuate carcinogenesis. We describe host porting the contribution of estrogen to prostate cancer, we genetic variations in the estradiol pathway associated sought to comprehensively determine if single-nucleotide with hormone levels and lethal form of prostate polymorphisms (SNP) in genes associated with estradiol- cancer. Single-nucleotide polymorphisms distributed related metabolic pathways, either individually or in com- across CYP1B1, HSD17B2, catechol-O-methyltransfer- bination, would be predictors of prostate cancer progression. ase (COMT), and SULT2B1 were associated with pro- We established germline genetic signature in these pathways gression and survival. Indeed, based on our findings, associated with cancer progression in localized prostate patients could be stratified into four distinct prognostic cancers and then verified these findings in locally advanced subgroups. Thus, the cumulative impact of such markers prostate cancers to evaluate the influence of the signature on provides clinically relevant information from a single overall survival. blood sample, is invariant with time, overcomes tumor heterogeneity, and identifies potential targets to opti- Patients and Methods mize disease management. Our data offer a high trans- Clinical data lational potential that could lead to a personalize The study included 739 Caucasian patients with prostate approach based on the presence or absence of the cancer divided into two independent cohorts. The first molecular signature and identify novel promising drug- cohort was composed of 526 patients with localized pros- gable targets. tate cancer, and the second was composed of 213 patients with locally advanced disease. Patients had undergone surgical radical prostatectomies at l’Hotel-Dieu^ de Quebec Hospital (Quebec City, Canada) before being enrolled in hormones (13, 14) and that these pathways are dysregu- the study. The localized and advanced prostate cancer lated in prostate cancer (15). Moreover, the consequence of cohorts were recruited between 1999 and 2002, and sustained and active steroidogenesis in cancer cells is 1982 and 2002, respectively (5). The median follow-up highlighted by the observation that castration-resistant times were 7.4 and 7.8 years for the localized and locally prostate cancer can still be driven by sex steroid hormones advanced populations, respectively. Before surgery, each despite a low circulating testosterone (T) level (14). Several participant provided written consent for genetic analysis. germline polymorphisms in sex steroid metabolic pathways The local research ethics committee approved the research have been shown to be associated with localized and protocol. advanced prostate cancer outcomes related to a meaningful influence of inherited genetic variations in the androgenic Genetic analysis pathway on progression (4, 8). Therefore, in addition to DNA was extracted from peripheral blood mononuclear tumor markers, circulating tumor cells, and whole-blood cells obtained at time of diagnosis. For genotyping, PCR gene-expression profiles (2, 3, 16), host polymorphisms amplification was performed on germline DNA, and the might assist in predicting tumor behavior and serve as a products were sequenced by Sequenom iPLEX matrix- simple and reliable assay easily analyzed from germline assisted laser desorption/ionization–time-of-flight
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