Contribution of CMV-specific T cells to chronic kidney rejection

CMV 特异性 T 细胞对慢性肾排斥反应的影响

• 批准号: 10054619
• 负责人: Lauren Higdon
• 金额: $ 15.41万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-08 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10054619
• 关键词: Acceleration; Address; Adult; Affect; Age; Aging; Antibodies; Antiviral Agents; Archives; Biological Assay; Biopsy; Biopsy Specimen; Blood; CD8-Positive T-Lymphocytes; Cell Aging; Cells; Characteristics; Chromatin; Chronic; Clinical; Collaborations; Complication; Computer Analysis; Control Groups; Core Facility; Cytomegalovirus; Cytomegalovirus Infections; Data; Data Analyses; Data Set; Detection; Development; Diagnosis; Elderly; End stage renal failure; Environment; Exposure to; Feasibility Studies; Flow Cytometry; Frequencies; Gene Expression Profiling; Generations; Goals; Graft Survival; Health; Imaging Techniques; Immune; Immune response; Immunity; Immunosuppression; Impairment; Incidence; Individual; Infection; Infiltration; Inflammation; Inflammatory; Kidney; Kidney Diseases; Kidney Transplantation; Knowledge; Learning; Length; Life; Maintenance; Measures; Mediating; Microscopy; Morbidity - disease rate; Opportunistic Infections; Organ Transplantation; Outcome; Pathogenicity; Pathologist; Patients; Peptides; Peripheral Blood Mononuclear Cell; Phenotype; Population; Predisposition; Process; Production; Prophylactic treatment; Proteins; Renal Cell Carcinoma; Research; Research Personnel; Risk; Role; Sampling; Savings; Scientist; Signal Transduction; Site; Sorting - Cell Movement; Stains; Surface; Symptoms; T memory cell; T-Lymphocyte; Testing; Time; Training; Transplant Recipients; Transplantation; Viral; Viremia; Virus Latency; Virus Replication; aged; aging population; base; career; cell age; cohort; complex data; cytokine; cytotoxic; cytotoxicity; experience; immunosenescence; improved; indexing; innovation; kidney allograft; kidney biopsy; kidney dysfunction; macrophage; mortality; mortality risk; novel diagnostics; novel therapeutics; post-transplant; post-transplant disease; premature; receptor; response; senescence; seropositive; single-cell RNA sequencing; telomere; transcription factor

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.

项目概要 肾移植对于终末期肾病 (ESKD) 患者来说可以挽救生命。然而，并发症 包括机会性感染以及慢性 T 细胞和抗体介导的排斥反应，减少长期移植物 生存。巨细胞病毒（CMV）血清阳性与术后慢性排斥反应的较高发生率相关。 移植。长期接触 CMV 会诱导 CMV 反应性记忆 T 细胞发生免疫衰老， 包括增殖能力和分化潜力的丧失，同时维持炎症的产生 细胞因子。结果，这些细胞的保护作用减弱，甚至可能促进排斥反应。我的 初步数据表明，CMV 反应后衰老的发展加速。 移植。此外，T细胞介导的炎症巨噬细胞浸润至肾移植物中可以 诱发排斥反应。巨噬细胞是CMV感染的主要部位。这些观察结果共同导致了 衰老的 CMV 反应性 T 细胞会导致慢性排斥反应的假设将得到测试 通过两个目标。第一个目标是表征 CMV 反应性 T 细胞的衰老特征 移植。 CMV 反应性 T 细胞将在移植后时间点进行分析 加速衰老。表征将包括单细胞 RNA 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。目标是 第二个目的是确定 CMV 反应性 T 细胞和炎症巨噬细胞是否影响 对慢性排斥反应的易感性。 CMV 特异性 T 细胞和 CMV 特异性 T 细胞浸润的数量和定位 将在存档的活检标本中测量进入同种异体肾移植物的巨噬细胞，以比较患者 诊断为慢性排斥反应的对照组为慢性排斥反应患者。血T表型分析 移植活检时的细胞将识别与排斥相关的群体，从长远来看 可能会提供新的排斥反应诊断。 PI的职业规划是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、统计学家合作获得专业知识的机会， 肾病学家和病理学家，这是职业规划的关键。具体来说，目标 1 涉及与 Purvesh Khatri 博士负责测序数据集的复杂数据分析。目标 2 涉及与 移植肾病专家 Jane Tan 博士和 Paul Grimm 博士以及病理学家 Neeraja Kambham 博士进行分析和 活检结果的解释。它还涉及与肾细胞癌研究人员 Wendy 博士的合作 Fantl 和统计学家 Robert Tibshirani 博士对活检标本进行了高度多重分析。这些目标是 旨在了解 CMV 反应性 T 细胞在慢性排斥反应中的作用，可能有助于 开发针对慢性排斥的新疗法，并支持申请人向独立过渡。