Contribution of CMV-specific T cells to chronic kidney rejection

项目摘要 肾移植是终末期肾病（ESKD）患者的救命之道。然而，并发症 包括机会性感染和慢性T细胞和抗体介导的排斥反应， 生存巨细胞病毒（CMV）血清阳性与术后慢性排斥反应的发生率较高相关。 移植慢性暴露于CMV诱导CMV应答性记忆T细胞的免疫衰老， 包括增殖能力和分化潜能的丧失，同时维持炎性细胞的产生， 细胞因子结果，这些细胞的保护性降低，甚至可能促进排斥反应。我 初步数据表明，衰老的发展，响应CMV加速后， 移植此外，T细胞介导的炎性巨噬细胞浸润到移植肾中， 诱导排斥。巨噬细胞是CMV感染的主要部位。总之，这些观察结果导致了 将检验衰老CMV应答性T细胞导致慢性排斥的假设 通过两个目标。第一个目标的目标是表征CMV应答性T细胞在施用后的衰老。 移植CMV应答性T细胞将在移植后时间点在CMV感染发作之前和之后进行分析。 加速衰老。表征将包括通过单细胞RNA进行的深入基因表达谱分析 染色质可及性、端粒长度和增殖的测序和功能测定。的目标 第二个目的是确定CMV应答性T细胞和炎性巨噬细胞是否影响 对慢性排斥反应的敏感性。CMV特异性T细胞和巨噬细胞浸润的数量和定位 将在存档的活检标本中测量巨噬细胞进入同种异体肾移植物的情况， 以确诊为慢性排斥反应的慢性患者为对照组。血T细胞表型分析 移植活检时的细胞将识别与排斥相关的群体， 可能提供一种新的排斥诊断方法。职业计划是让PI成为一名独立调查员 研究免疫并发症对肾移植的影响。斯坦福大学的研究环境是 非常适合这些研究，可以使用测序、流式细胞术和显微镜的核心设施。 这些目标提供了与生物信息学家，统计学家， 肾科医生和病理学家，这是职业规划的关键。具体而言，目标1涉及与 博士Purvesh Khatri用于测序数据集的复杂数据分析。目标2涉及与 移植肾学家Jane Tan博士和Paul Grimm博士以及病理学家Neeraja Kambham博士进行分析， 活检结果的解释。它还涉及与肾细胞癌研究人员温迪博士的合作 Fantl和统计学家Robert Tibshirani博士对活检标本进行高度多重分析。这些目的是 制定以了解CMV反应性T细胞在慢性排斥反应中的作用， 为慢性排斥反应提供新的治疗方法，并支持申请人向独立过渡。

Project Summary Kidney transplantation is life-saving for people with end stage kidney disease (ESKD). However, complications including opportunistic infection and chronic T cell- and antibody-mediated rejection reduce long-term graft survival. Seropositivity for cytomegalovirus (CMV) is associated with higher rates of chronic rejection after transplantation. Chronic exposure to CMV induces immunosenescence in CMV-responsive memory T cells, including loss of proliferative capacity and differentiation potential, with maintained production of inflammatory cytokines. In consequence, these cells become less protective, and may even promote rejection. My preliminary data suggest that the development of senescence in response to CMV is accelerated after transplantation. In addition, T cell mediated infiltration of inflammatory macrophages into kidney grafts can induce rejection. Macrophages are a major site of CMV infection. Together, these observations led to the hypothesis that senescent CMV-responsive T cells contribute to chronic rejection that will be tested through two aims. The goal of the first aim is to characterize senescence in CMV-responsive T cells post- transplant. CMV-responsive T cells will be analyzed at post-transplant time points before and after the onset of acceleration of senescence. Characterization will include in-depth gene expression profiling by single cell RNA sequencing and functional assays for chromatin accessibility, telomere length, and proliferation. The goal of the second aim is to determine whether CMV-responsive T cells and inflammatory macrophages influence susceptibility to chronic rejection. Quantity and localization of infiltration of both CMV-specific T cells and macrophages into kidney allografts will be measured in archived biopsy specimens, comparing patients diagnosed with chronic rejection to a control group of patients with chronicity. Phenotypic analysis of blood T cells at the time of transplant biopsy will identify populations correlated with rejection, which in the long-term may provide a new diagnostic of rejection. The career plan is for the PI to become an independent investigator researching the effects of immune complications on kidney transplant. The research environment at Stanford is well suited for these studies, given access to core facilities for sequencing, flow cytometry, and microscopy. The aims provide opportunities to gain expertise in collaboration with bioinformaticians, statisticians, nephrologists, and pathologists, which are key to the career plan. Specifically, Aim 1 involves collaboration with Dr. Purvesh Khatri for complex data analysis of sequencing data sets. Aim 2 involves collaboration with transplant nephrologists Drs Jane Tan and Paul Grimm and pathologist Dr. Neeraja Kambham for analysis and interpretation of biopsy results. It also involves collaboration with renal cell carcinoma researcher Dr. Wendy Fantl and statistician Dr. Robert Tibshirani for highly multiplexed analysis of biopsy specimens. These aims are formulated to gain understanding of the role of CMV-responsive T cells in chronic rejection, potentially to produce new therapies to chronic rejection, and to support the applicant’s transition to independence.