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

免疫系统既抗击癌症，又抗击感染。癌症免疫疗法试图利用和增强这种能力。FDA批准的一种免疫疗法涉及修改患者的白细胞以识别和攻击肿瘤细胞。产生的细胞被称为CAR-T细胞。了解CAR-T活性可以使改进的癌症免疫疗法的设计成为可能。确定在CAR-T细胞中增强肿瘤杀伤的信号通路是该项目的主要目标。这项研究项目将整合更广泛的教育计划，以教育学生、教师和公众。对当地一所高中的多年外展将为以西班牙裔为主的学生群体提供实践经验。嵌合抗原受体(CARS)由模块化的蛋白结构域组成，包括细胞内信号/共刺激结构域。CAR激活刺激细胞内的磷酸化级联反应，从而促进T细胞的细胞毒功能。磷酸蛋白质组学提供了蛋白质磷酸化的定量特征。主要的假设是，使用磷酸蛋白质组学对CAR信号进行严格和全面的研究将揭示CAR功能的原理。这可能使下一代汽车的理性设计成为可能。使用CD19作为模型抗原，该项目将尝试扩大汽车信号和汽车设计标准的知识。该项目将追求以下科学目标：1)确定促进CAR-T细胞细胞毒功能的信号通路；2)确定能够抵抗疲劳的CAR信号通路；3)开发具有更高细胞毒性和更高耐用性的第三代汽车。这项研究将建立一个可用于汽车信号工程分析的平台，并将提供汽车设计的总体框架。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势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.