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: 9976049
• 负责人: Zachary Ryan Healy
• 金额: $ 12.51万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9976049
• 关键词: Advisory Committees; Antigens; Antiviral Response; Apoptosis; Area; Automobile Driving; Award; Biological Assay; Biology; Cell Death; Cell Therapy; Cells; Cellular Metabolic Process; Characteristics; Clinical; Complex; Complication; Cytolysis; Cytomegalovirus; Cytomegalovirus Infections; Data; Dependence; Ensure; Event; Flow Cytometry; Funding; Future; Generations; Genetic Transcription; Glutamine; Goals; Immunocompromised Host; Immunology; Immunosuppression; Inflammatory; Institution; Interleukin-2; Interleukin-7; Investigation; JAK2 gene; Kidney Transplantation; Laboratory Research; Leadership; Maintenance; Measures; Mentorship; Metabolic; Metabolic Pathway; Metabolism; Mitochondria; Molecular; Morbidity - disease rate; Organ Transplantation; Pathway interactions; Patients; Phenotype; Physicians; Polyamines; Positioning Attribute; Prevention therapy; Production; Protocols documentation; RNA; Reproducibility; Research; Risk; Risk Assessment; Role; Scientist; Signal Transduction; Solid; Stat5 protein; T cell response; T-Cell Activation; T-Lymphocyte; T-Lymphocyte Subsets; Technical Expertise; Techniques; Testing; Therapeutic; Time; Training; Transcript; Transplant Recipients; Transplantation; Up-Regulation; Viral; allograft rejection; antigen-specific T cells; base; career; cost; cytokine; cytotoxicity; design; differential expression; exhaustion; high risk; improved; metabolic profile; mortality; novel; post-transplant; prevent; response; skills; tenure track; tool; transcriptome sequencing; 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），由此产生 在增加同种异移植排斥和死亡率中。当前的病毒学研究表明，抗原的存在 特定的“多功能” T细胞对于病毒控制至关重要。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）并影响快速的细胞溶解， 而那些仅表达一个（即单功能）或没有细胞因子的细胞与 逐渐减少保护。不幸的是，定量多功能T细胞的测定仍然很艰巨 并且缺乏可重复性的间隔性；此外，推动生成和维护的机制 这些细胞仍然未知。我在肾脏移植接受者中的初步数据表明，那些缺乏的人 多功能CMV响应的CMV重新激活风险增加。重要的是，我还发现 细胞耗尽并不是观察到与CMV相关功能差异的主要负责 在有和没有CMV重新激活的患者之间，因此提出了替代机制。所以， 基于初步数据，所检验的中央假设是STAT5-和MYC依赖性分子 重新编程对于生成和维护CMV特异性多功能T细胞，以及 用基于IL-7的协议扩展细胞的细胞概括了此重编程，并将产生 在CMV重新激活的情况下，功能性，长期基于细胞的疗法。拟议的研究将：（1） 确定是否可以使用转录曲线来量化多功能T细胞，还可以识别 SOT接受者有CMV重新激活的风险； （2）确定STAT5和MYC激活的关键作用以及 CMV特异性多功能T细胞功能的替代代谢途径表达； （3） 通过表征来优化基于细胞因子的方案，以实现CMV特异性T细胞的离体扩展 功能，增殖和代谢参数。我的主要职业目标是获得终身职位 并在主要的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 建立作为独立资助的身体科学家的职业，特别关注移植 T细胞代谢的免疫学和相关变化。为了实现这些目标，我将发展我的 智力基础，增强我的领导能力，并通过 拟议研究的持续时间。 DRS的实验室很容易获得宝贵的培训。默多克（Murdoch），混乱和 Piantadosi。此外，为了在奖励期间促进和促进我的进度，我已经组织了一个 具有专业知识的良好科学家和临床医生的科学咨询委员会在所有领域的所有领域都有专业知识 应用。总的来说，拟议的研究将增强我们对特定分子的理解 应对移植受者中多功能T细胞的产生和维护的机制，以及 可能会为具有CMV重新激活的移植受者提供更好的治疗选择。 呢 呢 呢