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

前列腺癌从雄激素依赖到去势抵抗的机制 前列腺癌（CRPC），以及介导对靶向残留雄激素治疗的耐药性的那些 CRPC中的AR受体（AR）信号传导，包括第二代AR拮抗剂如Enzalutamide， 被充分阐明。包括我们自己在内的几个小组的工作都强调了 CRPC对AR的影响，并暗示EZH 2是参与AR重编程的额外致癌驱动因子， CRPC。在本资助期内完成的研究中，我们探索了EZH 2作为靶点， CRPC的模型，并且已经证明催化EZH 2抑制剂在这些模型中具有活性。目标 该提案的主要目的是利用新的基因组规模的CRISPR/Cas9和ORF筛选技术， 确定激素独立性和敏感性的基本基因及其功能， CRPC中对EZH 2抑制剂、AR拮抗剂和其他靶向药物的耐药性。此外，在合作 与Freedman实验室合作，我们将使用CRISPR/Cas9编辑顺式调控元件来详细探索 一种新的体细胞获得性转录增强子在控制AR基因表达中的作用 在CRPC。我们还将确定由ARv 7和其他AR驱动的CRPC模型中的必需基因， 与Project 2合作的突变体和变体。最后，我们将进行CRISPR和ORF筛选， 鉴定潜在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.