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

T细胞在骨髓中产生，在胸腺中成熟。它们能抵御包括癌症在内的多种疾病。有些癌症会伪装自己不被t细胞识别。另一些则抑制t细胞的杀伤反应。T细胞可以用嵌合抗原受体（CARs）修饰。这些合成分子为T细胞提供了一种新的感知和摧毁癌细胞的能力。CAR修饰的T细胞在临床上对对抗一些血癌是有效的。不幸的是，我们对调节T细胞如何对癌细胞作出反应的参数的理解严重缺乏。该项目旨在获得对调节T细胞反应的CAR参数的基本理解，并表征这些参数如何影响CAR工程T细胞的功能和命运。该研究项目与一项教育计划相结合，该计划旨在支持坦帕湾地区社区学院系统的学生，为学生在STEM道路上导航时提供指导和指导研究经验。嵌合抗原受体（CARs）是一种模拟T细胞受体激活和共刺激的合成T细胞受体。这些经验设计的受体提供了一种有效但未优化的信号反应。因此，经过改造的T细胞容易受到过度激活或抑制的影响，这是细胞疗法面临的两种常见但不同的挑战。诸如去磷酸化动力学和下游适配器偏好等重要参数对CAR信号传导的影响基本上尚未得到研究。优化受体功能以支持稳健的T细胞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car的信号传导成分如何在激活和抑制条件下产生更好反应的基本理解。这些实验将通过以下途径提供基础进展：1)研究合成T细胞活化过程中磷酸化、去磷酸化和适配器结合的基序景观；2)定量了解去磷酸化动力学如何影响CAR信号传导；3)利用适配器偏置定位T细胞中的合成信号。该奖项反映了美国国家科学基金会的法定使命，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。

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.