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

确定肾细胞癌中 DNA 复制应激反应缺陷的免疫学后果，以改善免疫治疗结果

• 批准号: 10640407
• 负责人: Daniel James McGrail
• 金额: $ 24.9万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-03 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10640407
• 关键词: Advisory Committees; Area; Biological Markers; CRISPR screen; Candidate Disease Gene; Cells; Clear cell renal cell carcinoma; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Combined Modality Therapy; Cytometry; DNA; DNA Damage; DNA biosynthesis; Defect; Early Diagnosis; Ensure; Environment; Etiology; Faculty; Gene Deletion; Gene Expression; Gene Expression Profile; Genes; Genetic Engineering; Genetically Engineered Mouse; Genotype; Goals; Immune; Immune response; Immunobiology; Immunologics; Immunologist; Immunology; Immunooncology; Immunotherapy; Knowledge; Lead; Link; Location; Malignant Neoplasms; Mediating; Medical Oncology; Mentors; Mentorship; Modality; Modeling; Mus; Mutation; Neoplasm Transplantation; Outcome; Patient-Focused Outcomes; Patients; Pharmaceutical Preparations; Pharmacology; Phase; Population; Production; Proteins; Quality of life; Recording of previous events; Renal Cell Carcinoma; Renal carcinoma; Research Personnel; Resistance; Role; STING1 gene; Sampling; Screening Result; Signal Transduction; Single-Stranded DNA; Stimulator of Interferon Genes; System; Testing; Training; Tumor-infiltrating immune cells; Validation; base; biological adaptation to stress; career; clinical biomarkers; cohort; computational pipelines; cytokine; genetic signature; high throughput screening; immune checkpoint blockade; improved; improved outcome; in vivo; insight; malignant breast neoplasm; member; mouse model; multiplexed imaging; neoplastic cell; novel therapeutics; overexpression; patient stratification; patient subsets; predicting response; predictive marker; programmed cell death ligand 1; replication stress; response; responsible research conduct; side effect; skills; success; targeted treatment; tenure track; tool; transcriptome sequencing; transplant model; tumor; tumor-immune system interactions

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.

项目总结/摘要 虽然免疫疗法在肾细胞癌（RCC）中显示出强大的临床反应，但只有 三分之一的患者会受益。目前的临床生物标志物，如突变负荷或PD-L1表达， 证明在RCC中没有预测能力;然而，申请人最近鉴定了一种基因签名， DNA复制应激反应（RSR）缺陷可预测多种免疫治疗反应， 独立的群体。该提案将测试RSR缺陷导致STING激活的中心假设 信号传导以促进对免疫检查点阻断敏感性，以及RSR的药理学诱导 缺陷可使原本具有抗性的肿瘤对免疫疗法敏感。在K99辅导阶段， 申请人将使用高度多重成像质谱细胞术来分析免疫细胞中的差异， RSRD低和RSRD高患者之间的微环境（目标1.1）。然后，申请人将生成一个 多种基因工程RCC小鼠移植模型，以实现受控的体内研究，以及 验证RSR缺陷与免疫治疗反应存在因果关系（目标1.2）。在目标2中，申请人将 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高模型（Aim 2.1）介导的STING缺失和体内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）。申请人将过渡到R 00 在CRISPR筛选完成后的独立阶段。CRISPR筛选结果可以作为研究的基础。 第一次R 01提交。目标3将寻求鉴定将诱导RSR缺陷的药理学化合物， 使其他抗性模型对免疫疗法敏感，并将作为第二次R 01应用的基础。 这项提议汇集了一个由在DNA损伤方面具有专业知识的非凡研究人员组成的咨询委员会， 免疫肿瘤学、肾细胞癌医学肿瘤学、基因工程小鼠模型和高- 通过筛选，以加强申请人在这些领域的技能。这将通过课程作业得到补充 在免疫学和负责任的研究行为，以及额外的培训，以准备申请人 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方之一。总的来说，这项建议将 为申请人提供工具，以启动职业生涯作为终身教职员工，同时提供关键的 深入了解RCC的免疫治疗，以推进MD安德森的“让癌症成为历史”的目标。