Coactivators and Androgen Receptors in Prostate Cancer

前列腺癌中的共激活剂和雄激素受体

• 批准号: 7057211
• 负责人: Nancy L. Weigel
• 金额: $ 33.07万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-01 至 2008-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7057211
• 关键词: androgen receptor; biological signal transduction; carcinogenesis; cell growth regulation; gene expression; genetic promoter element; genetic regulation; genetic transcription; hormone regulation /control mechanism; immunoprecipitation; laboratory mouse; ligands; male; posttranslational modifications; prostate neoplasms; receptor binding; receptor expression

DESCRIPTION (provided by applicant): Prostate cancer is initially an androgen dependent disease and there is mounting evidence that the androgen receptor (AR) continues to play a role in androgen ablation resistant prostate cancer. Great strides have been made in recent years in elucidating the structure of AR, identifying candidate coregulatory proteins, and in identifying some of the genes regulated by AR. Nonetheless, our understanding of the mechanism of action of AR and our knowledge of the identity of the key genes regulated by AR to induce growth of prostate cancer cells is insufficient to provide a good basis for rational development of alternate means of blocking AR activity. Our understanding of and ability to effectively block the growth of androgen independent prostate cancer is even less well advanced. Although many candidate AR coactivators have been identified by molecular biological techniques, virtually nothing is known about their individual roles in cells, their contributions to positive and negative regulation of AR target genes and to AR dependent cell growth. Moreover, their role in androgen independent AR action has not been determined. To address these issues, we propose: Specific Aim 1: To test the hypothesis that the androgen receptor plays a key role in the growth and gene expression in some androgen independent prostate cancer cells as well as in androgen dependent prostate cancer cells and to evaluate the role of receptor coactivators in both hormone dependent and independent cell growth and gene regulation. We will selectively reduce expression of candidate coactivators and assess the effects on cell growth and target gene expression. Specific Aim 2: To identify genes regulated by androgen receptor and to test the hypothesis that coactivator requirements are target gene/promoter specific. We will utilize two approaches, Affymetrix U133A chips and a novel Protein A-AR chimera containing a TEV (tobacco etch virus) protease site linker to identify AR target genes and to evaluate the consequences of coactivator reduction on target gene expression. Specific Aim 3: To utilize chromatin immunoprecipitation (CHIP) assays to identify coregulators involved in hormone dependent and ligand independent transcriptional regulation by androgen receptors and to assess the consequences of elimination of specific coregulators on the composition of proteins associated with promoters and on the post-translational modifications of proteins associated with the promoter.

描述（由申请人提供）： 前列腺癌最初是一种依赖雄激素的疾病，并且有越来越多的证据表明，雄激素受体（AR）继续在抗雄激素消融耐药性前列腺癌中发挥作用。近年来，在阐明AR的结构，鉴定候选蛋白质以及鉴定AR调节的某些基因方面取得了长足的进步。但是，我们对AR作用机理的理解以及我们对AR调节的关键基因的身份的了解，以诱导前列腺癌细胞的生长，不足以为阻止AR活性的替代方法的合理发展提供良好的基础。我们对有效阻止雄激素独立前列腺癌生长的理解和能力的进步甚至不太好。尽管已经通过分子生物学技术鉴定了许多候选AR共激活剂，但实际上对它们在细胞中的各个角色，它们对AR靶基因的正和阴性调节的贡献以及对AR依赖性细胞生长的贡献几乎一无所知。此外，尚未确定它们在雄激素独立AR作用中的作用。为了解决这些问题，我们提出：具体目的1：检验以下假设：雄激素受体在某些独立的雄激素独立前列腺癌细胞以及依赖雄激素的前列腺癌细胞中在生长和基因表达中起关键作用，并评估受体共同激活剂在激素依赖性和独立细胞生长和基因调节中的受体共同激活者的作用。我们将有选择地减少候选共激活因子的表达，并评估对细胞生长和靶基因表达的影响。具体目的2：确定受雄激素受体调节的基因，并测试共激活因素是靶基因/启动子特异性的假设。我们将利用两种方法：Affymetrix U133A芯片和一种新型的蛋白A-AR嵌合体，其中含有TEV（烟草蚀刻病毒）蛋白酶位点接头，以鉴定AR靶基因并评估共激活剂降低靶基因表达的后果。具体目的3：利用染色质免疫沉淀（芯片）测定来鉴定与激素依赖性和配体的抗激素依赖性和配体无关的转录调节，并评估消除与启动子以及与蛋白质后以及与蛋白质相关的蛋白质相关的特定组分剂对蛋白质组成的影响。