Determining the Immunological Consequences of DNA Replication Stress Response Defects in Renal Cells Carcinoma to Improve Immunotherapy Outcomes

项目摘要/摘要 虽然免疫疗法在肾癌(RCC)的临床反应中显示出巨大的前景，但只有 每3名患者中就有1名将受益。目前的临床生物标记物，如突变负荷或PD-L1表达， 被证明在肾癌中没有预测能力；然而，申请人最近发现了一个基因签名 DNA复制应激反应(RSR)缺陷可预测多发性硬化患者的免疫治疗反应 独立的队列。这一提议将检验RSR缺陷导致STING激活的中心假设 增强对免疫检查点阻断敏感性信号，以及RSR的药理诱导 缺陷可以使原本具有抗药性的肿瘤对免疫疗法敏感。在K99指导阶段， 申请者将使用高度多元化的成像质量细胞术来分析免疫差异 RSRD-Low和RSRD-High患者之间的微环境(目标1.1)。然后申请者将生成一个 多种基因工程RCC小鼠移植模型，以实现体内受控研究，以及 确认RSR缺陷与免疫治疗反应有因果关系(目标1.2)。在目标2中，申请者将 寻求从机制上理解RSR缺陷如何通过CRISPR促进免疫治疗反应- 从RSRD-HIGH模型(AIM 2.1)介导的STING删除和体内CRISPR筛选靶向基因 包含在RSRD签名中，以确定因果关系(目标2.2)。申请者将过渡到R00 CRISPR屏幕完成后的独立阶段。CRISPR筛查结果可以作为 第一份R01提交。目标3将寻求识别会导致RSR缺陷的药理化合物 使其他耐药模型对免疫疗法敏感，并将作为第二次R01应用的基础。 这项提议汇集了一个由在DNA损伤方面拥有专业知识的现象级研究人员组成的咨询委员会， 免疫肿瘤学、肾细胞癌内科肿瘤学、基因工程小鼠模型和高密度脂蛋白 吞吐量筛选，以增强申请者在这些领域的技能。这将由课程作业来补充。 在免疫学和负责任的研究方面，以及为申请人做好准备的额外培训 领导一个独立的实验室。MD安德森的环境，包括研究人员和设施， 使这里成为少数几个可以完成这个项目的地点之一。综上所述，这项提议将 为申请者配备工具，使其能够以终身教职员工的身份启动工作，同时提供关键的 深入了解免疫疗法治疗肾癌，以推进MD Anderson创造癌症历史的目标。

PROJECT SUMMARY/ABSTRACT While immunotherapy has shown great promise for robust clinical responses in renal cell carcinoma (RCC), only 1 in 3 patients will see benefit. Current clinical biomarkers, such as mutation burden or PD-L1 expression, have proven to have no predictive capacity in RCC; however, the applicant has recently identified a gene signature of DNA replication stress response (RSR) defects that has predicted immunotherapy response in multiple independent cohorts. This proposal will test the central hypothesis that RSR defects lead to activation of STING signaling to promote sensitivity to immune checkpoint blockade, and that pharmacological induction of RSR defects can sensitize otherwise resistant tumors to immunotherapy. During the K99 mentored phase, the applicant will use highly multiplexed imaging mass cytometry to analyze differences in the immune microenvironment between RSRD-low and RSRD-high patients (Aim 1.1). The applicant will then generated a diverse array of genetically engineered RCC mouse transplant models to enable controlled in vivo studies, and validate that RSR defects are causally linked to immunotherapy response (Aim 1.2). In Aim 2, the applicant will seek to mechanistically understand how RSR defects contribute to immunotherapy response by CRISPR- mediated deletion of STING from an RSRD-high model (Aim 2.1) and in vivo CRISPR screens targeting genes contained within the RSRD signature to ascertain causality (Aim 2.2). The applicant will transition to the R00 independent phase as the CRISPR screen is completed. The CRISPR screen results can serve as a basis for a first R01 submission. Aim 3 will seek to identify pharmacological compounds that will induce RSR defects to sensitize otherwise resistant models to immunotherapy, and will serve as the basis for a second R01 application. This proposal brings together an advisory committee of phenomenal researchers with expertise in DNA damage, immuno-oncology, renal cell carcinoma medical oncology, genetically engineered mouse models, and high- throughput screening to bolster the applicant's skillset in these areas. This will be supplemented by coursework in immunology and responsible conduct of research, as well as additional training to prepare the applicant for leading an independent lab. The environment at MD Anderson, including both the researchers and facilities, makes this one of the few locations where this project could be accomplished. Taken together, this proposal will equip the applicant with the tools to launch by career as a tenure-track faculty member while providing critical insight into immunotherapy for treatment of RCC to advance MD Anderson's goal of Making Cancer History.