Characterization of CMV-specific T cell responses in immunocompromised hosts

免疫受损宿主中 CMV 特异性 T 细胞反应的表征

• 批准号: 10700143
• 负责人: Zachary Ryan Healy
• 金额: $ 24.59万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-07 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10700143
• 关键词: Advisory Committees; Antigens; Area; Automobile Driving; Award; Biological Assay; Cell Therapy; Cells; Cellular Metabolic Process; Complication; Cytolysis; Cytomegalovirus; Data; Event; Funding; Gene Expression Profile; Generations; Goals; IL7 gene; Immunocompromised Host; Inflammatory; Institution; Interleukin-2; Kidney Transplantation; Laboratory Research; Leadership; Maintenance; Metabolic; Metabolic Pathway; Molecular; Organ Transplantation; Patients; Physicians; Positioning Attribute; Protocols documentation; Reproducibility; Research; Risk; Role; Scientist; Stat5 protein; T cell response; T-Lymphocyte; Technical Expertise; Testing; Training; Transplant Recipients; Transplantation Immunology; Viral; allograft rejection; base; career; cytokine; exhaustion; mortality; response; skills; tenure track; virology

The most common infectious complication following organ transplantation is cytomegalovirus (CMV), resulting in increased allograft rejection and mortality. Current virology research suggests that the presence of antigen- specific “polyfunctional” T cells is essential for viral control; when encountering target, these rare cells are capable of producing multiple inflammatory cytokines (e.g., IFNg, TNFa, IL-2) and effecting rapid cytolysis, while those cells expressing only one (i.e., monofunctional) or no cytokines have been associated with progressively less protection. Unfortunately, assays for quantification of polyfunctional T cells remains arduous and lack inter-facility reproducibility; additionally, the mechanisms driving the generation and maintenance of such cells remains unknown. My preliminary data in kidney transplant recipients reveals that those lacking polyfunctional CMV responses are at increased risk of CMV reactivation. Importantly, I have also found that T cell exhaustion is not primarily responsible for discrepancies in CMV-associated functionality observed between patients with and without CMV reactivation, thereby suggesting alternative mechanisms. Therefore, based on preliminary data, the central hypothesis tested is that STAT5- and myc-dependent molecular reprogramming is necessary for the generation and maintenance of CMV-specific polyfunctional T cells, and that expansion of cells with an IL-7-based protocol recapitulates this reprogramming and will produce functional, long-lived cell-based therapies in the event of CMV reactivation. The proposed research will: (1) determine if a transcriptional profile may be used not only to quantify polyfunctional T cells but also identify SOT recipients at risk for CMV reactivation; (2) determine the critical role of STAT5 and myc activation and alterative metabolic pathways expression on the function of CMV-specific polyfunctional T cells; and (3) optimize cytokine-based protocols for the ex-vivo expansion of CMV-specific T cells by characterization of functional, proliferative, and metabolic parameters. My primary career goal is to obtain a tenure-track position and establish an independent research laboratory at a major biomedical institution. My long-term career goal is to establish a career as an independently-funded physician-scientist, with a particular focus in transplant immunology and associated changes in T cells metabolism. To achieve these goals, I will develop my intellectual base, strengthen my leadership skills, and enhance the necessary technical skills through the duration of the proposed study. Valuable training is readily available in the labs of Drs. Murdoch, Chao, and Piantadosi. Furthermore, to promote and bolster my progress during the award period, I have organized a scientific advisory committee of well-established scientists and clinicians with expertise in all areas of the application. Collectively, the proposed research will enhance our understanding of the specific molecular mechanisms regulating the generation and maintenance of polyfunctional T cells in transplant recipients, and may result in better treatment options for transplant recipients with CMV reactivation.

器官移植后最常见的感染并发症是巨细胞病毒(CMV)，导致 增加了同种异体移植排斥反应和死亡率。目前的病毒学研究表明，抗原的存在- 特殊的“多功能”T细胞对于病毒控制是必不可少的；当遇到目标时，这些稀有细胞是 能够产生多种炎性细胞因子(例如，IFNG、TNFa、IL-2)并实现快速的细胞溶解， 而那些只表达一种(即，单功能)或不表达细胞因子的细胞与 保护力度越来越小。不幸的是，对多功能T细胞进行量化的分析仍然很困难 并且缺乏设施间的重复性；此外，驱动产生和维护 这样的细胞仍然未知。我在肾移植受者中的初步数据显示，那些缺乏 多功能巨细胞病毒应答增加了巨细胞病毒重新激活的风险。重要的是，我还发现T 细胞耗尽不是观察到的CMV相关功能差异的主要原因 在有和没有CMV重新激活的患者之间，从而提出了替代机制。因此， 根据初步数据，检验的中心假设是STAT5和myc依赖的分子 重新编程对于CMV特异性多功能T细胞的产生和维持是必要的，并且 使用基于IL-7协议的细胞扩展概括了这种重新编程，并将产生 在巨细胞病毒重新激活的情况下，基于细胞的功能性、长寿疗法。拟议的研究将：(1) 确定转录图谱是否不仅可以用来量化多功能T细胞，还可以用来识别 面临CMV重新激活风险的SO受体；(2)确定STAT5和myc激活的关键作用和 改变代谢途径对CMV特异性多功能T细胞功能的影响 优化基于细胞因子的CMV特异性T细胞体外扩增方案 功能、增殖和代谢参数。我的主要职业目标是获得终身教职的职位 并在一家大型生物医学机构建立独立的研究实验室。我的长期职业目标是 建立一个独立资助的内科科学家的职业生涯，特别是移植方面的重点 免疫学和T细胞代谢的相关变化。为了实现这些目标，我将发展我的 知识基础，加强我的领导能力，并通过 拟议研究的持续时间。有价值的培训在默多克博士、赵小兰博士和 皮安塔多西。此外，为了促进和支持我在获奖期间的进步，我组织了一个 由在各个领域都有专长的知名科学家和临床医生组成的科学咨询委员会 申请。总的来说，拟议的研究将增强我们对特定分子的理解 调节移植受者多功能T细胞的产生和维持的机制，以及 对于CMV重新激活的移植受者，可能会有更好的治疗选择。