Project 4: Identification of Essential Genes Underlying AR Activity in Antagonist-Resistant CRPC

前列腺癌从雄激素依赖性进展到去势抵抗的机制 前列腺癌(CRPC)，以及那些对以残留雄激素为靶点的治疗产生抵抗的患者 CRPC中的受体(AR)信号转导包括第二代AR拮抗剂，如苯扎鲁胺 得到了充分的阐明。包括我们自己在内的几个组织的研究突显了这种持续的依赖 并暗示EZH2是参与AR重新编程的另一个致癌驱动因素 CRPC。在当前资助期间完成的研究中，我们将EZH2作为#年的目标 CRPC模型，并已证明催化EZH2抑制剂在这些模型中具有活性。目标是 这项建议的目的是利用新的基因组规模的CRISPR/Cas9和ORF筛选技术来 确定构成激素独立性和敏感性的基本基因及其功能 CRPC对EZH2抑制剂、AR拮抗剂和其他靶向药物的耐药性。此外，在协作中 通过弗里德曼实验室，我们将使用CRISPR/Cas9编辑顺式法规元素来详细探索 一种新的体细胞获得性转录增强子对AR基因表达的调控作用 它本身在CRPC中。我们还将定义ARv7和其他AR驱动的CRPC模型中的必要基因 突变体和变种与项目2合作。最后，我们将进行CRISPR和ORF筛查 确定潜在的新的合成致死组合和抗性机制，包括对一组新的 与项目3合作开发的激酶抑制剂。

The mechanisms underlying prostate cancer progression from androgen dependent to castration resistant prostate cancer (CRPC), as well as those mediating resistance to therapies that target the residual androgen receptor (AR) signaling in CRPC including second generation AR antagonists such as enzalutamide have not been fully elucidated. Work from several groups including our own has highlighted the continued dependence of CRPC on AR and has implicated EZH2 as an additional oncogenic driver involved in AR reprogramming in CRPC. In studies accomplished during the current funding period we have explored EZH2 as a target in models of CRPC and have demonstrated that catalytic EZH2 inhibitors have activity in these models. The goal of this proposal is to leverage the novel genome-scale CRISPR/Cas9 and ORF screening technology to identify the essential genes and their functions that underlie the hormone independence and sensitivity or resistance to EZH2 inhibitors, AR antagonists, and other targeted agents in CRPC. In addition, in collaboration with the Freedman lab we will use CRISPR/Cas9 editing of cis-regulatory elements to explore in detail the function of a novel somatically acquired transcriptional enhancer in controlling the expression of the AR gene itself in CRPC. We will also define the essential genes in models of CRPC driven by ARv7 and other AR mutants and variants in collaboration with Project 2. Finally, we will perform CRISPR and ORF screens to identify potential new synthetic lethal combinations and mechanisms of resistance including to a set of novel kinase inhibitors in collaboration with the Project 3.