Targeted disruption of the YAP/TAZ/TEAD axis in pancreatic cancer

项目总结/摘要 胰腺导管腺癌（PDAC）是胰腺癌的一种毁灭性形式， 结果。在PDAC中通常突变的KRAS是这种疾病中的驱动癌基因。然而，在这方面， 靶向KRAS的方法的开发已被证明具有挑战性， KRAS下游如MAPK途径的药物作为单一药物已经很大程度上失败。这个项目的总体目标是 一项建议是使用创新的化学策略来确定PDAC的联合方案，这是迫切需要的。 来提高患者的生存率。结合CRISPR筛选和表观基因组分析，我发现 雅普/TAZ/TEAD轴是KRAS缺失后旁路存活程序所需的关键转录节点 或MEK抑制。这些数据和已发表的文献支持了直接靶向 PDAC中的雅普/TAZ/TEAD轴。然而，目前的雅普/TAZ/TEAD抑制剂具有较差的效力和脱靶 研究关键转录调节因子的效果和遗传策略由于在转录调节因子的选择和选择之间的延迟而具有有限的效用。 蛋白质损失和实验测量。为了克服这些限制，我开发了一个通用的基于标签的 被称为降解标签（dTAG）系统的技术平台，用于诱导任何目标的快速降解 细胞系和小鼠模型中的蛋白质。dTAG系统使得能够以小的阈值来评估靶蛋白损失。 分子降解剂在一个时间尺度上，这是不可能的遗传方法，有利于评估突变 KRAS、雅普和TAZ。此外，格雷实验室开发了一种选择性小分子共价TEAD， 不可逆地抑制异常雅普/TAZ/TEAD信号传导。本申请中提出的工作将 利用dTAG技术平台和共价TEAD抑制剂来确定雅普/TAZ/TEAD在以下方面的作用： 在PDAC中协调旁路存活。在目标1中，在没有直接雅普或TAZ抑制剂的情况下，我将使用 dTAG系统来定义直接的雅普和TAZ转录信号传导程序，其促进细胞存活， KRAS信号传导的调节。这些实验将证明雅普的化学降解潜力 和TAZ，并确定新的有针对性的漏洞。在目标2中，我将使用PDAC细胞系和患者来源的 类器官模型，以评估共价TEAD抑制作为组合方案的翻译潜力 KRAS信号中断。整合化学生物学、基因组规模分析和翻译模型 的PDAC，我希望PDAC特定的治疗见解将从这项工作中产生。为了实现这些目标，我 制定了一项3年计划，其中包括参加科学和职业发展会议、讲习班和 课程，以进一步发展我的癌症化学生物学和计算生物学专业知识。这个职业 过渡奖将极大地促进我领导一个多学科研究实验室的目标，重点是 解决胰腺癌治疗中的挑战。

Project Summary/Abstract Pancreatic ductal adenocarcinoma (PDAC) is a devastating form of pancreatic cancer with dismal patient outcome. KRAS, which is commonly mutated in PDAC, is a driving oncogene in this disease. However, development of approaches to target KRAS has proven challenging and inhibitors targeting signaling networks downstream of KRAS such as the MAPK pathway have largely failed as single agents. The overall goal of this proposal is to use innovative chemical strategies to identify combination regimens in PDAC, which are urgently needed to improve patient survival. Combining CRISPR screens and epigenomic profiling, I identified that the YAP/TAZ/TEAD axis is a critical transcriptional node required for the bypass survival program upon KRAS loss or MEK inhibition. This data and published literature support the importance of directly targeting the YAP/TAZ/TEAD axis in PDAC. However, current YAP/TAZ/TEAD inhibitors have poor potency and off-target effects and genetic strategies to study critical transcription regulators have limited utility due to delays between protein loss and experimental measurement. To overcome these limitations, I developed a versatile tag-based technology platform known as the degradation tag (dTAG) system to induce rapid degradation of any target protein in cell lines and mouse models. The dTAG system enables evaluation of target protein loss with a small molecule degrader in a time-scale that is not possible with genetic approaches, facilitating evaluations of mutant KRAS, YAP and TAZ. In addition, the Gray laboratory developed a selective small molecule covalent TEAD inhibitor to irreversibly inactivate aberrant YAP/TAZ/TEAD signaling. The work proposed in this application will leverage the dTAG technology platform and a covalent TEAD inhibitor to establish the role of YAP/TAZ/TEAD in coordinating bypass survival in PDAC. In Aim 1, in the absence of direct YAP or TAZ inhibitors, I will use the dTAG system to define the direct YAP and TAZ transcriptional signaling program that promotes survival upon modulation of KRAS signaling. These experiments will demonstrate the potential of chemical degradation of YAP and TAZ and identify novel targetable vulnerabilities. In Aim 2, I will use PDAC cell lines and patient-derived organoid models to evaluate the translational potential of covalent TEAD inhibition as a combination regimen with KRAS signaling disruption. Integrating chemical biology, genome-scale analyses, and translational models of PDAC, I expect that PDAC-specific therapeutic insights will emanate from this work. To achieve these aims, I designed a 3-year plan that includes participation in scientific and career development meetings, workshops and coursework to further develop my cancer chemical biology and computational biology expertise. This career transition award will greatly facilitate my goal of leading a multidisciplinary research laboratory focused on addressing challenges in the treatment of pancreatic cancer.