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: 10609429
• 负责人: Alec Thomas McIntosh
• 金额: $ 3.5万
• 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/10609429
• 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; Phase; Phase I Clinical Trials; Phosphotransferases; Physical condensation; Protein Dephosphorylation; 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; combat; confocal imaging; epigenomics; genetic element; genetic signature; improved; in vivo; in vivo Model; industry partner; inhibitor; insight; mechanical signal; mutant; novel therapeutic intervention; novel therapeutics; pharmacologic; prevent; promoter; response; segregation; 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) 是美国第八大最常见的成人癌症。尽管最近 随着治疗选择的改善，大多数转移性肾细胞癌患者继续死于这种情况 疾病，每年导致全世界超过 100,000 人死亡。因此，寻找新的治疗方法非常重要 更有效地治疗这种致命疾病的策略。我的初步研究表明 YAP/TAZ 功能 作为核心致癌转录网络的主要调节因子，并且是维持 RCC 肿瘤的转录稳态。我的建议的第一个主要目标是确定以下措施的影响： YAP/TAZ 对致癌转录和超级增强子维持的药理和遗传抑制 在患者来源的原代肾细胞癌细胞中。我将采用尖端的表观基因组技术，包括 Cut&Tag 和 QuantSeq 探讨 YAP/TAZ 的遗传和药理学抑制如何影响组合物和 YAP/TAZ 转录复合物的活性以及整体 RCC 表观遗传和转录 景观。鉴于 YAP/TAZ 在协调相分离转录中的报道功能 超级增强子的凝结物，我将结合内源性 YAP/TAZ 的免疫荧光分析和 YAP/TAZ 转录复合物的其他成分，以及 DNA 荧光原位杂交 （DNA-FISH）针对 YAP/TAZ 结合的增强子区域，以研究遗传或药理学如何 YAP/TAZ 的抑制会影响这些特定基因的活性转录缩合物的形成 元素及其调节 YAP/TAZ 靶基因的必要性。通过这些分析，我将能够 深入了解 YAP/TAZ 在 RCC 肿瘤中的分子功能，同时重建核心 RCC 致癌转录网络的整体。该提案的第二个主要目标是确定 YAP/TAZ 如何与缺氧诱导因子 2 α (HIF2A) 功能性相互作用以及 YAP/TAZ 是否共同作用 在体内模型中，抑制可以克服对 HIF2A 抑制剂的耐药性。虽然HIF2A抑制剂已处于晚期 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很普遍。我的初步 研究结果表明，YAP/TAZ 信号传导可能有助于 HIF2A 抑制剂的耐药性。使用 YAP/TAZ 和 HIF2A 的药理学抑制剂，我打算使用 Cut&Tag 和 QuantSeq 来研究 HIF2A 和 YAP/TAZ 在维持透明细胞 RCC 致癌转录中的功能相互作用，如 以及可能的 YAP/TAZ 调节基因介导对 HIF2A 抑制剂的耐药性。此外，我将 研究 YAP/TAZ 抑制剂与 HIF2A 抑制剂联合使用如何影响 HIF2A-i- 的体内生长 抗性患者来源的 ccRCC 异种移植物，对收获的肿瘤进行 QuantSeq 以验证我的体外实验 发现。总之，这些研究将极大地增强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关系，可能产生更多 克服肾细胞癌患者治疗抵抗的有效联合策略。