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

项目 4：拮抗剂抗性 CRPC 中 AR 活性必需基因的鉴定

• 批准号: 10363641
• 负责人: MYLES A BROWN
• 金额: $ 37.05万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-05-24 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10363641
• 关键词: AR gene; Ablation; Androgen Antagonists; Androgen Receptor; Androgens; Antiandrogen Therapy; CRISPR interference; CRISPR library; CRISPR screen; CRISPR/Cas technology; Candidate Disease Gene; Castration; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Collaborations; Computer Models; Custom; Data; Dependence; EZH2 gene; Enhancers; Essential Genes; Funding; Generations; Genes; Genetic Transcription; Goals; Growth; Hormones; Individual; Knock-out; Libraries; Mediating; Modeling; Nucleic Acid Regulatory Sequences; Oncogenic; Open Reading Frames; Phenotype; Receptor Signaling; Regulatory Element; Research Project Grants; Residual state; Resistance; Structure; Technology; Validation; Variant; Work; Xenograft Model; advanced prostate cancer; androgen sensitive; antagonist; castration resistant prostate cancer; design; enzalutamide; epigenomics; experimental study; gain of function; genome wide screen; genome-wide; in vivo; inhibitor; kinase inhibitor; mutant; new therapeutic target; novel; prostate cancer cell line; prostate cancer model; prostate cancer progression; resistance mechanism; screening; small hairpin RNA; targeted agent; targeted treatment; therapy resistant

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.

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)，以及那些介导对针对残留雄激素的治疗​​产生耐药性的癌症 CRPC 中的受体 (AR) 信号转导，包括第二代 AR 拮抗剂，如恩杂鲁胺，尚未 得到了充分阐明。包括我们自己在内的多个小组的工作强调了持续的依赖性 CRPC 对 AR 的影响，并暗示 EZH2 作为参与 AR 重编程的额外致癌驱动因素 CRPC。在当前资助期间完成的研究中，我们探索了 EZH2 作为目标 CRPC 模型，并证明催化 EZH2 抑制剂在这些模型中具有活性。目标 该提案的目的是利用新型基因组规模的 CRISPR/Cas9 和 ORF 筛选技术 识别激素独立性和敏感性的基础基因及其功能，或 CRPC 对 EZH2 抑制剂、AR 拮抗剂和其他靶向药物的耐药性。此外，在合作中 我们将与 Freedman 实验室一起使用 CRISPR/Cas9 编辑顺式调控元件来详细探索 新型体细胞获得性转录增强子在控制 AR 基因表达中的功能 本身在 CRPC 中。我们还将定义 ARv7 和其他 AR 驱动的 CRPC 模型中的必需基因 与项目 2 合作开发突变体和变体。最后，我们将进行 CRISPR 和 ORF 筛选 确定潜在的新合成致死组合和耐药机制，包括一组新的 与项目 3 合作的激酶抑制剂。