Regulation of Androgen Receptor Conformation

描述(申请人提供)：雄激素受体(AR)是一种核受体，是前列腺癌(CAP)的主要治疗靶点。这项拨款涉及调节AR活性的细胞途径的特征，这可能产生新的治疗靶点，以及应用一种新的筛选策略来识别抑制AR功能的小分子。我们推测Rho/ROCK/LIM激酶(RRL)通路通过改变肌动蛋白细胞骨架来调节AR活性。我将使用稳定表达天然AR和荧光标记AR的前列腺癌株来确定RRL途径如何影响AR的折叠、二聚化、核输入和转录活性。我们还假设，抑制AR配体诱导的构象变化的化合物可能揭示出传统AR抑制剂筛选所遗漏的新的抗雄激素。我将使用高通量FRET分析来筛选三个具有生物活性的小分子集合，以寻找抑制AR配体诱导的构象变化的化合物。最有效的抑制剂将在详细的二次分析中进行检验，以确定它们对AR核运输、二聚化、天然AR反应转录和前列腺癌细胞增殖的影响。

DESCRIPTION (provided by applicant): The androgen receptor (AR) is a nuclear receptor that is the primary therapeutic target of prostate cancer (CaP). This grant concerns characterization of a cellular pathway that regulates AR activity, which could produce new therapeutic targets, and the application of a novel screening strategy to identify small molecules that inhibit AR function. We hypothesize that the Rho/ROCK/LIM kinase (RRL) pathway regulates AR activity through alterations in the actin cytoskeleton. I will use prostate carcinoma lines that stably express native and fluorescently-labeled AR to determine how the RRL pathway influences the folding, dimerization, nuclear import, and transcriptional activity of AR. We also hypothesize that compounds that inhibit AR ligand-induced conformational change may reveal novel anti-androgens that would be missed by traditional screens for AR inhibitors. I will use a high-throughput FRET assay to screen three collections of biologically active small molecules for compounds that inhibit AR ligand-induced conformational change. The most effective inhibitors will be examined in detailed secondary analyses to determine their effect on AR nuclear transport, dimerization, native AR-responsive transcription, and proliferation of prostate cancer cells.