Genome-Wide Analysis of Transcription Factor Function in Prostate Cancer

描述（由申请人提供）：雄激素受体（AR）是一种配体依赖性转录因子，在前列腺癌的发生和发展中起关键作用，是一种治疗靶点。令人惊讶的是，人们对人类基因组中AR结合、AR协同转录因子和AR靶基因的调控知之甚少。本提案的总体目标是从全基因组角度研究雄激素依赖性（AD）和非依赖性（Al）前列腺癌细胞中AR及其协同转录因子对蛋白质编码基因和一类非编码基因（microRNA [miRNA]）的组合转录调控。为了解决这些问题，我们将使用染色质免疫沉淀（ChIP）结合人类全基因组询问平片微阵列（ChlP-on-chip）来研究AD和Al前列腺癌中转录因子的体内结合及其调控功能。我们的具体目的是：(1)确定AD和Al前列腺癌细胞中是否存在不同的AR结合、AR协同转录因子伙伴和AR靶基因。AR chlp芯片检测将在AD和Al前列腺癌细胞中进行。AR结合及其协同转录因子和AR靶基因将通过生物信息学算法进行预测和实验验证。(2)确定AR及其协同转录因子如何在AD和Al前列腺癌细胞中联合调控AR靶基因。将执行协同转录因子ChlP-on-chip，并与AR ChlP-on-chip和基因表达谱相关联，以鉴定AD和Al前列腺癌细胞中AR靶基因的组合转录调控代码。(3)确定AR及其协同转录因子是否调控AD与Al前列腺癌细胞中的mirna。RNA聚合酶II (pol II) chlp芯片将在AD和Al前列腺癌细胞中进行。Pol II结合将与AR及其协同转录因子结合和miRNA表达谱相关，以确定AD和Al前列腺癌细胞中差异转录因子调节的miRNA表达。综上所述，这些研究将增加我们对AD和Al前列腺癌中AR及其协同转录因子在全基因组水平上对靶编码基因和非编码基因的差异转录调控的基本认识，从而为AD和Al前列腺癌的治疗干预寻找新的分子靶点。

DESCRIPTION (provided by applicant): The androgen receptor (AR), a ligand-dependent transcription factor, plays a key role in the onset and progression of prostate cancer and is a therapeutic target. Surprisingly little is known of AR binding, AR collaborating transcription factors, and regulation of AR target genes in the human genome. The overall goal of this proposal is to investigate the combinatorial transcriptional regulation of protein-coding genes and a class of non-coding genes (microRNA [miRNA]) by AR and its collaborating transcription factors from a genome-wide view in androgen-dependent (AD) and -independent (Al) prostate cancer cells. To address these issues, we will use chromatin immunoprecipitation (ChIP) combined with human whole genome interrogating tiling microarrays (ChlP-on-chip) to study in vivo binding of transcription factors and their regulatory function in AD and Al prostate cancer. Our specific aims are to: (1) Determine whether distinct AR binding, AR collaborating transcription factor partners and AR target genes exist in AD and Al prostate cancer cells. AR ChlP-on-chip assays will be performed in AD and Al prostate cancer cells. AR binding, its collaborating transcription factors and AR target genes will be predicted by bioinformatics algorithms and experimentally validated. (2) Determine how AR and its collaborating transcription factors combinatorially regulate AR target genes in AD and Al prostate cancer cells. Collaborating transcription factors ChlP-on-chip will be performed and correlated with AR ChlP-on-chip and gene expression profiles to identify combinatorial transcriptional regulatory codes for AR target genes in AD and Al prostate cancer cells. (3) Determine whether AR and its collaborating transcription factors regulate miRNAs in AD versus Al prostate cancer cells. RNA polymerase II (pol II) ChlP-on-chip will be performed in AD and Al prostate cancer cells. Pol II binding will be correlated to AR and its collaborating transcription factors bindings and miRNA expression profiles to identify differential transcription factors-regulated miRNA expression in AD and Al prostate cancer cells. In summary, these studies will increase our fundamental understanding of differential transcriptional regulation of target coding and non-coding genes by AR and its collaborating transcription factors on a genome-wide level in AD and Al prostate cancer, which will lead to identification of new molecular targets for therapeutic intervention in AD and Al prostate cancer.