Single-cell dissection of CD4 T cell changes in patients with immune-related adverse events following PD-1 inhibition

项目摘要 该申请提出了一个五年的研究和培训计划，重点是科学的机制 PD-1检查点抑制导致免疫相关不良事件（irAE）。这个假设是 PD-1信号转导的从头缺失增强了CD 4 T细胞的抗原敏感性，并导致CD 4 T细胞的活化。 自身反应性CD 4 T细胞介导irAE的发展。为了研究这个假设，候选人将 分析他的导师的两个互补的人类队列：a）PD-1患者的前瞻性irAE队列 免疫治疗，以比较发生（irAE+）和未发生（irAE-）的患者中CD 4 T细胞的早期变化 irAE，和B）接受季节性流感疫苗接种以测试的PD-1免疫疗法的患者队列 irAE+患者中是否存在基础免疫病理学，在基础免疫学评价中未观察到 在免疫的帮助下，可以被发现。使用前瞻性irAE队列，目标1将评价 发生irAE的患者是否具有不同的初始CD 4 T细胞表观遗传和转录特征， 基线，其允许PD-1抑制后自身反应性CD 4 T细胞的稳健活化。此外，这一目标将 分析PD-1抑制后产生的活化CD 4 T细胞的转录谱和克隆多样性 在irAE+患者中。使用流感疫苗接种的aPD-1队列，目标2将回答以下问题： PD-1信号传导是微调TCR阈值和CD 4 T细胞应答的转录谱所必需的。 维持免疫力和体内平衡。这些研究将确定PD-1抑制导致 自身反应性CD 4 T细胞的活化和增殖引起免疫介导的病理学。他们将 还确定了可用于早期诊断和治疗患有 风湿病和其他irAE。从科学角度来说，候选人的职业发展目标是获得专业知识 在临床和转化研究，生物信息学分析，单细胞技术，以及 患者样品中自身免疫途径的转录和表观遗传调节。从理论上讲， 候选人旨在获得科学写作，拨款准备，沟通和数据展示方面的经验 最终提交R 01。为了促进候选人作为一名物理学家和科学家的成长， 结合了新颖的实验性单细胞方法和特定的职业发展计划， 候选人和他的导师，威利博士和劳弗博士。候选人的长期目标是过渡到任期- 跟踪教师的立场，并制定一个独立的NIH资助的研究计划，重点是机制 自身免疫和T细胞功能障碍，以最终预防和治疗风湿性疾病。丰富的科学 和合作环境在宾夕法尼亚大学将定位候选人有一个高度 在人类自身免疫方面有影响力的转化研究事业。

Project Summary This application proposes a five-year research and training plan with a scientific focus on the mechanisms through which PD-1 checkpoint inhibition leads to immune-related adverse events (irAEs). The hypothesis is that de novo loss of PD-1 signaling enhances the antigen sensitivity of CD4 T cells and leads to the activation of autoreactive CD4 T cells that mediate irAE development. To investigate this hypothesis, the candidate will analyze two complementary human cohorts of his mentors: a) a prospective irAE cohort of patients on PD-1 immunotherapy to compare the early changes of CD4 T cells in patients who do (irAE+) and do not (irAE-) develop irAEs, and b) a cohort of patients on PD-1 immunotherapy that receive seasonal influenza vaccination to test whether any underlying immunopathology in irAE+ patients, not otherwise observed with evaluation of basal states, can be revealed with the help of immunization. Using the prospective irAE cohort, Aim 1 will evaluate whether patients that develop irAEs have a different naïve CD4 T cell epigenetic and transcriptional profile at baseline that allows robust activation of autoreactive CD4 T cells following PD-1 inhibition. Further, this aim will analyze the transcriptional profile and clonal diversity of the activated CD4 T cells generated after PD-1 inhibition in irAE+ patients. Using the aPD-1 cohort of influenza vaccination, Aim 2 will answer the question of whether PD-1 signaling is required to finetune the TCR threshold and transcriptional profile of CD4 T cell responses to maintain immune fitness and homeostasis. These studies will define the path through which PD-1 inhibition leads to the activation and propagation of autoreactive CD4 T cells to cause immune-mediated pathology. They will also identify cellular and molecular targets that can be used for early diagnosis and treatment of patients with rheumatologic and other irAEs. Scientifically, the candidate’s career development goals are to gain expertise in the conduct of clinical and translational research, bioinformatic analyses, single-cell technologies, and transcriptional and epigenetic regulation of autoimmune pathways in patient samples. Professionally, the candidate aims to gain experience in scientific writing, grant preparation, communication, and data presentation culminating to an R01 submission. To facilitate the candidate’s growth as a physician-scientist, this proposal combines novel experimental single-cell approaches and a specific career development plan designed by the candidate and his mentors, Dr. Wherry and Dr. Laufer. The candidate’s long-term goal is to transition to a tenure- track faculty position and to develop an independent NIH-funded research program focusing on the mechanisms of autoimmunity and T cell dysfunction to ultimately prevent and treat rheumatologic diseases. The rich scientific and collaborative environment at the University of Pennsylvania will position the candidate to have a highly impactful, translational research career in human autoimmunity.