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

项目摘要 这项申请提出了一个五年的研究和培训计划，科学地关注机制 通过PD-1检查点抑制导致免疫相关不良事件(IrAEs)。假设是 PD-1信号的从头丢失增强了CD4T细胞的抗原敏感性，导致了CD4T细胞的活化 介导IRAE发育的自身反应性CD4T细胞。为了调查这一假设，候选人将 分析他的导师的两个互补的人类队列：a)服用PD-1的预期患者队列 免疫治疗比较(IRAE+)和未发展(IRAE-)患者CD4T细胞的早期变化 IrAEs，以及b)接受季节性流感疫苗接种以进行测试的PD-1免疫治疗患者队列 在IRAE+患者中是否有任何潜在的免疫病理学，没有通过基础评估进行观察 在免疫接种的帮助下，可以发现。使用未来的IRAE队列，AIM 1将评估 发生irAEs的患者是否有不同的幼稚CD4T细胞表观遗传学和转录图谱 基线，允许在PD-1抑制后强健地激活自身反应性CD4T细胞。此外，这一目标将 PD-1抑制后活化的CD4T细胞的转录谱和克隆多样性分析 在IRAE+患者中。使用apd-1流感疫苗接种队列，目标2将回答以下问题 需要PD-1信号来微调TCR阈值和CD4T细胞反应的转录图谱 保持免疫健康和动态平衡。这些研究将确定PD-1抑制的途径 导致自身反应性CD4T细胞的激活和增殖，引起免疫介导的病理改变。他们会 还要确定可用于早期诊断和治疗的细胞和分子靶点 风湿病和其他irAEs。科学地说，候选人的职业发展目标是获得专业知识 在临床和转化性研究、生物信息学分析、单细胞技术以及 患者样本中自身免疫途径的转录和表观遗传调节。在专业方面， 应聘者的目标是获得科学写作、拨款准备、沟通和数据展示方面的经验 最终提交了R01。为了促进候选人作为内科科学家的成长，这项提议 结合了新颖的实验性单细胞方法和由 候选人和他的导师惠里博士和劳弗博士。候选人的长期目标是过渡到任期-- 跟踪教职员工的位置，并制定一个独立的由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病。富有的科学人 宾夕法尼亚大学的协作环境将使候选人具有高度的 在人类自身免疫方面有影响力的转译研究生涯。

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.