CAREER: Engineering next-generation chimeric antigen receptors for cancer immunotherapy using phospho-proteomics

免疫系统对抗癌症和感染。癌症免疫疗法试图利用和加强这种能力。FDA批准的一种免疫疗法涉及修改患者的白色血细胞以识别和攻击肿瘤细胞。由此产生的细胞被称为CAR-T细胞。了解CAR-T活性可以设计改进的癌症免疫疗法。确定增强CAR-T细胞中肿瘤杀伤的信号通路是该项目的主要目标。这个研究项目将整合一个更广泛的教育计划，以教育学生，教师和公众。多年的推广到当地高中将提供实践经验，以西班牙裔学生为主。嵌合抗原受体（汽车）由模块化蛋白结构域组成，包括细胞内信号传导/共刺激结构域。CAR活化刺激促进T细胞的细胞毒性功能的细胞内磷酸化级联。磷酸化蛋白质组学提供蛋白质磷酸化的定量表征。主导假设是使用磷酸蛋白质组学对CAR信号传导进行严格和全面的研究将揭示CAR功能的原理。这可以实现下一代汽车的合理工程。使用CD 19作为模型抗原，该项目将尝试扩展CAR信号传导和CAR设计标准的知识。该项目将追求以下科学目标：1）识别促进CAR-T细胞细胞毒性功能的信号通路; 2）定义能够抵抗耗竭的CAR信号通路; 3）开发具有改善的细胞毒性和增加的持久性的第三代汽车。这项研究将建立一个可用于CAR信号工程分析的平台，并将为CAR设计提供一个通用框架。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The immune system fights cancer as well as infections. Cancer immunotherapy attempts to harness and bolster this capability. One FDA-approved immunotherapy involves modifying white blood cells from the patient to recognize and attack tumor cells. The resulting cells are referred to as CAR-T cells. Understanding CAR-T activity could enable design of improved cancer immunotherapy. Identifying the signaling pathways that enhance tumor killing in CAR-T cells is a primary objective of the project. This research project will integrate a broader educational plan to educate students, teachers, and the public. Multi-year outreach to a local high school will provide hands-on experience to a predominantly Hispanic student population. Chimeric antigen receptors (CARs) are comprised of modular protein domains including intracellular signaling/co-stimulation domains. CAR activation stimulates intracellular phosphorylation cascades that promote the cytotoxic functions of T cells. Phospho-proteomics offers quantitative characterization of protein phosphorylation. The governing hypothesis is that a rigorous and comprehensive investigation of CAR signaling using phospho-proteomics will uncover principles of CAR function. This could enable rational engineering of next-generation CARs. Using CD19 as a model antigen, this project will attempt to expand knowledge of CAR signaling and CAR design criteria. The project will pursue the following scientific objectives: 1) Identify signaling pathways that promote CAR-T cell cytotoxic function; 2) Define CAR signaling pathways that enable resistance to exhaustion; and 3) Develop third-generation CARs with improved cytotoxicity and increased persistence. This research will establish a platform that can be used for engineering analysis of CAR signaling and will provide a general framework for CAR design.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.