Biological Studies

项目总结： 项目2的最终目标是发现驱动前列腺癌发病机制的基因。自.以来 到目前为止发现的大多数风险变异都位于已知蛋白质编码区之外(即内含子和 基因间)，需要系统化的方法来揭示风险等位基因和基因之间的联系 它所影响的。我们正在全面测试的主要假设是，风险区域 尚未发现的转录本和/或风险区域是功能元件(例如，启动子和 增强剂)影响基因表达。提出了一系列补充技术，将 彻底解决这些假设。将对前列腺转录组进行测序，以得出无偏倚的 编码和非编码转录本和替代剪接的普查。中与功能相关的元素 前列腺癌高危区域将使用染色质免疫沉淀和 DNA酶过敏症。最后，将使用各种方法来从功能上表征每个风险部位， 包括染色体构象捕捉，以识别所有其他与 候选基因的风险区域、基因扰动(例如，敲除和过表达)实验 围绕风险区域，评估风险等位基因和转录本丰度之间的关联 所有候选基因。了解导致前列腺癌的生物途径提供了可靠的基础 合理干预疾病防治。

Project Summary: The ultimate goal of project 2 is the discovery of the genes that drive prostate cancer pathogenesis. Since most of the risk variants discovered to date reside outside of known protein coding regions (i.e., intronic and intergenic), systematic approaches are required to reveal the connection between the risk allele and the gene that it influences. The primary hypotheses that we are comprehensively testing are that the risk regions harbor an as yet undiscovered transcript and/or that the risk regions are functional elements (e.g., promoters and enhancers) that influence gene expression. A range of complementary techniques is proposed that will thoroughly address these hypotheses. The prostate transcriptome will be sequenced to derive an unbiased census of coding and non-coding transcripts and alternative splicing. Functionally relevant elements within the prostate cancer risk loci regions will be annotated using the techniques of chromatin immunoprecipitation and DNase hypersensitivity. Finally, methods will be employed to functionally characterize each of the risk loci, including chromosome conformation capture to identify all other genomic regions that are interacting with the risk region, gene perturbation (e.g., knockdown and overexpression) experiments of candidate genes surrounding the risk regions, and evaluating associations between the risk alleles and transcript abundance across candidate genes. Understanding the biologic pathways driving prostate cancer provides a sound basis for rational intervention in disease prevention and treatment.