Biological Studies

项目概要： 项目 2 的最终目标是发现驱动前列腺癌发病机制的基因。自从 迄今为止发现的大多数风险变异位于已知的蛋白质编码区之外（即内含子和 基因间），需要系统的方法来揭示风险等位基因和基因之间的联系 它的影响。我们正在全面测试的主要假设是风险区域包含 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 增强子）影响基因表达。提出了一系列补充技术， 彻底解决这些假设。前列腺转录组将被测序以获得公正的结果 编码和非编码转录本以及选择性剪接的普查。内功能相关的元素 将使用染色质免疫沉淀技术注释前列腺癌风险位点区域 DNase 超敏反应。最后，将采用方法来对每个风险位点进行功能表征， 包括染色体构象捕获，以识别与相互作用的所有其他基因组区域 风险区域、候选基因的基因扰动（例如敲低和过度表达）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 跨候选基因。了解驱动前列腺癌的生物学途径提供了坚实的基础 以便合理干预疾病的预防和治疗。

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.