CAREER: Understanding the Impact of Dephosphorylation Kinetics and Adapter Specificity on Synthetic T Cell Receptor Signaling and Function

职业：了解去磷酸化动力学和接头特异性对合成 T 细胞受体信号传导和功能的影响

• 批准号: 2339172
• 负责人: Lawrence Stern
• 金额: $ 63.61万
• 依托单位: University of South Florida
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2024
• 资助国家: 美国
• 起止时间: 2024-02-01 至 2029-01-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2339172&HistoricalAwards=false
• 关键词: CAREER; Understanding; Impact; Dephosphorylation; Kinetics

T cells are created in bone marrow and mature in the thymus. They defend against a variety of diseases, including cancer. Some cancers disguise themselves from T-cell recognition. Others suppress the killing response of T-cells. T cells can be modified with chimeric antigen receptors (CARs). These are synthetic molecules that provide T cells with a renewed ability to sense and destroy cancer cells. CAR engineered T cells are clinically effective at fighting some blood cancers. Unfortunately, our understanding of parameters that tune how a T cell responds to a cancer cell is severely lacking. This project aims to gain a fundamental understanding of the parameters of CARs that tune T cell responses, and to characterize how these parameters affect CAR engineered T cell function and fate. This research program is integrated with an education plan that aims to support students in the community college system in the Tampa Bay area, providing mentorship and guided research experiences for students as they navigate the STEM pathway.Chimeric antigen receptors (CARs) are synthetic T cell receptors that mimic T cell receptor activation and co-stimulation. These empirically designed receptors provide a potent but unoptimized signaling response. As a result, engineered T cells are susceptible to overactivation or suppression, two common but distinct challenges to cell therapy. The influence of important parameters such as dephosphorylation kinetics and downstream adapter preferences on CAR signaling remains essentially unstudied. Optimizing receptor function to support robust T cell activation remains a trial-and-error exercise. This project will combine protein engineering, cell biology, and omics experiments to establish a fundamental understanding of how the signaling components of CARs can be tuned to yield improved responses under activating and suppressive conditions. These experiments will provide fundamental advances through 1) surveying motif landscapes for phosphorylation, dephosphorylation, and adapter binding in synthetic T cell activation; 2) quantitatively understanding how dephosphorylation kinetics influence CAR signaling; 3) mapping synthetic signaling in T cells with adapter bias.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.

T 细胞在骨髓中产生并在胸腺中成熟。 它们可以防御多种疾病，包括癌症。有些癌症会伪装自己以躲避 T 细胞的识别。其他药物则抑制 T 细胞的杀伤反应。 T 细胞可以用嵌合抗原受体 (CAR) 进行修饰。这些合成分子可为 T 细胞提供感知和破坏癌细胞的新能力。 CAR 工程 T 细胞在临床上可有效对抗某些血癌。不幸的是，我们对调节 T 细胞如何响应癌细胞的参数的理解严重缺乏。该项目旨在对调节 T 细胞反应的 CAR 参数有一个基本的了解，并描述这些参数如何影响 CAR 工程化 T 细胞的功能和命运。该研究项目与一项教育计划相结合，旨在支持坦帕湾地区社区大学系统的学生，为学生在 STEM 途径中提供指导和指导研究经验。嵌合抗原受体 (CAR) 是合成 T 细胞模拟 T 细胞受体激活和共刺激的受体。这些根据经验设计的受体提供了有效但未经优化的信号反应。 因此，工程化 T 细胞容易过度激活或抑制，这是细胞治疗面临的两个常见但又不同的挑战。去磷酸化动力学和下游接头偏好等重要参数对 CAR 信号传导的影响基本上尚未研究。优化受体功能以支持 T 细胞的强大激活仍然是一个反复试验的过程。该项目将结合蛋白质工程、细胞生物学和组学实验，以建立对如何调整 CAR 信号成分以在激活和抑制条件下产生改善反应的基本了解。这些实验将通过以下方面提供根本性进展：1) 调查合成 T 细胞激活中磷酸化、去磷酸化和接头结合的基序景观； 2）定量了解去磷酸化动力学如何影响CAR信号传导； 3) 绘制具有适配器偏差的 T 细胞中的合成信号。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。