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，在胸腺中成熟。 它们可以抵御各种疾病，包括癌症。有些癌症会伪装自己，不让T细胞识别。其他抑制T细胞的杀伤反应。T细胞可以用嵌合抗原受体（汽车）修饰。这些是合成分子，为T细胞提供了新的感知和破坏癌细胞的能力。CAR工程化T细胞在临床上有效地对抗一些血液癌症。不幸的是，我们对调节T细胞如何对癌细胞做出反应的参数严重缺乏了解。该项目旨在从根本上了解调节T细胞反应的汽车参数，并表征这些参数如何影响CAR工程化T细胞的功能和命运。该研究项目与旨在支持坦帕湾区社区学院系统学生的教育计划相结合，为学生在STEM途径中导航提供指导和指导研究经验。嵌合抗原受体（汽车）是模拟T细胞受体激活和共刺激的合成T细胞受体。这些凭经验设计的受体提供了有效但未优化的信号传导应答。 因此，工程化T细胞容易过度活化或抑制，这是细胞治疗的两个常见但不同的挑战。重要参数如去磷酸化动力学和下游衔接子偏好对CAR信号传导的影响基本上尚未研究。优化受体功能以支持强大的T细胞活化仍然是一种试错练习。该项目将结合联合收割机蛋白质工程，细胞生物学和组学实验，以建立一个基本的理解，如何可以调整汽车的信号成分，以产生激活和抑制条件下的改善反应。这些实验将通过1）调查合成T细胞活化中磷酸化、去磷酸化和衔接子结合的基序景观; 2）定量地理解去磷酸化动力学如何影响CAR信号传导;第三章该奖项反映了NSF的法定使命，并被认为值得通过使用基金会的知识产权进行评估来支持。优点和更广泛的影响审查标准。