The Oncogenic functions of YAP/TAZ in Renal Cell Carcinoma

YAP/TAZ 在肾细胞癌中的致癌功能

• 批准号: 10464566
• 负责人: Alec Thomas McIntosh
• 金额: $ 3.41万
• 依托单位: GEORGETOWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-07 至 2027-04-06
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10464566
• 关键词: Address; Adult; Affect; Binding; CRISPR interference; CRISPR/Cas technology; Cell Line; Cell Nucleus; Cessation of life; Clear cell renal cell carcinoma; Clinical; Clinical Trials; Complex; DNA; Data; Data Set; Development; Disease; Drug Targeting; Enhancers; Epigenetic Process; Fluorescent in Situ Hybridization; Generations; Genes; Genetic; Genetic Transcription; Genomics; Growth; Harvest; Homeostasis; Immune checkpoint inhibitor; Immunofluorescence Immunologic; In Vitro; Literature; Maintenance; Malignant Epithelial Cell; Malignant Neoplasms; Mediating; Metastatic Renal Cell Cancer; Modeling; Molecular; Mutate; Nutrient; Oncogenic; Oncoproteins; Oral; Output; Pathogenesis; Pathway interactions; Patients; Pharmacology; Phase; Phase I Clinical Trials; Phosphotransferases; Regulatory Element; Renal Cell Carcinoma; Reporting; Resistance; Resolution; Scaffolding Protein; Signal Transduction; Site; TAZ gene; Techniques; Technology; Testing; Therapeutic; Tissue Microarray; Tissues; Treatment outcome; United States; Xenograft procedure; bHLH-PAS factor HLF; base; combat; confocal imaging; epigenetic silencing; epigenomics; genetic element; genetic signature; improved; in vivo; in vivo Model; industry partner; inhibitor; insight; mechanical signal; mutant; novel therapeutic intervention; novel therapeutics; prevent; promoter; response; small molecule inhibitor; targeted treatment; therapy resistant; transcription factor; treatment effect; tumor; tumor microenvironment

PROJECT ABSTRACT Renal cell carcinoma (RCC) is the 8th most common form of adult cancer in the US. Despite recent improvement in treatment options, the majority of patients with metastatic RCC continue to succumb to this disease, resulting in over 100,000 deaths per year worldwide. Thus, it is important to identify novel therapeutic strategies to more effectively treat this deadly disease. My preliminary studies suggest that YAP/TAZ function as master regulators of the core oncogenic transcriptional network and are required for maintaining the transcriptional homeostasis of RCC tumors. The first major aim of my proposal is to determine the effects of pharmacologic and genetic inhibition of YAP/TAZ on oncogenic transcription and super enhancer maintenance in patient-derived primary RCC cells. I will employ cutting-edge epigenomic techniques including Cut&Tag and QuantSeq to probe how genetic and pharmacological inhibition of YAP/TAZ affects the compositions and activities of the YAP/TAZ transcriptional complexes and the global RCC epigenetic and transcriptional landscape. Given the reported functions of YAP/TAZ in orchestrating phase-separated transcriptional condensates at super enhancers, I will combine immunofluorescence analysis of endogenous YAP/TAZ and other components of the YAP/TAZ transcriptional complexes, along with DNA fluorescent in situ hybridization (DNA-FISH) against YAP/TAZ-bound enhancer regions, to investigate how genetic or pharmacological inhibition of YAP/TAZ affects the formation of active transcriptional condensates at these particular genetic elements and their necessity in regulating YAP/TAZ target genes. Through these analyses, I will be able to gain critical insights into the molecular functions of YAP/TAZ in RCC tumors, but also reconstruct the core ensemble of the RCC oncogenic transcriptional network. The second major aim of this proposal is to determine how YAP/TAZ functionally interacts with hypoxia-inducible factor 2 alpha (HIF2A) and whether YAP/TAZ co- inhibition can overcome resistance to HIF2A inhibitors in an in vivo model. While HIF2A inhibitors are in late stage clinical trials, upfront and acquired resistance to this small molecule inhibitor is prevalent. My preliminary findings suggest that YAP/TAZ signaling may contribute to resistance to HIF2A inhibitors. Using pharmacological inhibitors of YAP/TAZ and HIF2A, I intend to use Cut&Tag and QuantSeq to investigate the functional interactions between HIF2A and YAP/TAZ in maintaining clear cell RCC oncogenic transcription, as well as possible YAP/TAZ regulated genes mediating resistance to HIF2A inhibitors. Furthermore, I will investigate how YAP/TAZ inhibitors, in combination with HIF2A inhibitors, impact the in vivo growth of HIF2A-i- resistant patient-derived ccRCC xenografts, performing QuantSeq on harvested tumors to validate my in vitro findings. Together, these studies will greatly enhance our understanding of the crosstalk and redundancies between the oncogenic pathways that govern the growth and survival of RCC cells, potentially yielding more effective combination strategies to overcome treatment resistance in RCC patients.

项目摘要 肾细胞癌（RCC）是美国第8种最常见的成人癌症。尽管最近 随着治疗选择的改善，大多数转移性RCC患者继续死于这种疾病。 这种疾病每年在全世界造成超过10万人死亡。因此，重要的是确定新的治疗方法， 更有效地治疗这种致命疾病的策略。我的初步研究表明，雅普/TAZ功能 作为核心致癌转录网络的主要调节因子， RCC肿瘤的转录稳态。我的建议的第一个主要目的是确定 雅普/TAZ对致癌基因转录和超级增强子维持的药理学和遗传学抑制 在患者来源的原代RCC细胞中。我将采用尖端的表观基因组技术，包括切割和标记， QuantSeq以探测雅普/TAZ的遗传和药理学抑制如何影响组合物， 雅普/TAZ转录复合物的活性和整体RCC表观遗传和转录 景观鉴于报道的雅普/TAZ在协调分相转录中的功能， 在超级增强子处的浓缩物，我将结合内源性雅普/TAZ的联合收割机免疫荧光分析和 雅普/TAZ转录复合物的其他组分，沿着DNA荧光原位杂交 （DNA-FISH）针对雅普/TAZ结合的增强子区域，研究遗传或药理学 雅普/TAZ的抑制影响这些特定遗传位点活性转录缩合物的形成。 元件及其在调控雅普/TAZ靶基因中的必要性。通过这些分析，我将能够 获得对雅普/TAZ在RCC肿瘤中的分子功能的重要见解，但也重建了核心 RCC致癌转录网络的整体。该提案的第二个主要目的是确定 雅普/TAZ与缺氧诱导因子2 α（HIF 2A）在功能上如何相互作用，以及雅普/TAZ是否与HIF 2A共同作用。 抑制可以克服体内模型中对HIF 2A抑制剂的抗性。虽然HIF2A抑制剂在晚期 临床试验阶段，对这种小分子抑制剂的前期和获得性耐药性是普遍的。我的初步 研究结果表明，雅普/TAZ信号可能有助于对HIF 2A抑制剂的耐药性。使用 作为雅普/TAZ和HIF 2A的药理学抑制剂，我打算使用Cut & Tag和QuantSeq来研究 HIF 2A和雅普/TAZ在维持透明细胞RCC致癌转录中的功能相互作用， 以及可能的雅普/TAZ调节基因介导对HIF 2A抑制剂的抗性。此外，我将 研究雅普/TAZ抑制剂与HIF 2A抑制剂组合如何影响HIF 2A-i的体内生长。 耐药患者来源的ccRCC异种移植物，对收获的肿瘤进行QuantSeq，以验证我的体外 调查结果。总之，这些研究将大大提高我们对串扰和冗余的理解 控制RCC细胞生长和存活的致癌途径之间的联系，可能产生更多的 有效的联合策略，以克服RCC患者的治疗耐药性。