High dimensional single-cell mapping to define immune signatures of cytomegalovirus-associated rejection in cardiac transplantation

高维单细胞图谱定义心脏移植中巨细胞病毒相关排斥反应的免疫特征

• 批准号: 10369726
• 负责人: Pritha Sen
• 金额: $ 17.28万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10369726
• 关键词: Activated Lymphocyte; Advisory Committees; Allografting; Animal Model; Antigen Presentation; Antigens; Area; Automobile Driving; Award; Bioinformatics; Biological Markers; Biology; Biopsy; Blood; Cardiac; Cell Communication; Cells; Cellular Indexing of Transcriptomes and Epitopes by Sequencing; Clonal Expansion; Cytomegalovirus; Cytomegalovirus Infections; Cytotoxic T-Lymphocytes; Data; Development; Development Plans; Disease; Endothelial Cells; Epithelial; Epitopes; Flow Cytometry; Fostering; Foundations; Funding; General Hospitals; Genetic Transcription; Goals; Graft Rejection; Heart; Heart Transplantation; Human; Human Subject Research; Immune; Immune System Diseases; Immunity; Immunofluorescence Microscopy; Immunology; Immunophenotyping; Immunosuppression; In Situ Hybridization; Infiltration; Inflammasome; Inflammatory; Interdisciplinary Study; Ligands; Longevity; Major Histocompatibility Complex; Massachusetts; Measurement; Mediating; Membrane Proteins; Memory; Mentorship; Methods; Molecular; Mononuclear; Morbidity - disease rate; Opportunistic Infections; Organ; Outcome; Phagocytes; Pharmaceutical Preparations; Phenotype; Physicians; Population; Precipitation; Production; Proteins; Proteomics; Research; Research Personnel; Resolution; Risk; Risk Factors; Role; Scientist; System; T cell receptor repertoire sequencing; T-Cell Receptor; T-Lymphocyte Subsets; Testing; Tissues; Training; Transplant Recipients; Transplantation; Viral; Viral Cytopathogenic Effect; Viral reservoir; Viremia; Virus Diseases; Work; allograft rejection; career development; chemokine; cohort; cytotoxic; design; diagnostic tool; heart allograft; high dimensionality; high risk; immunological synapse; in vivo; isoimmunity; multiple omics; novel; pathogen; precision medicine; receptor; response; shift work; single-cell RNA sequencing; trafficking; translational study; transplant centers; treatment strategy

PROJECT SUMMARY Human cytomegalovirus (CMV) is the most common viral infection to complicate cardiac transplantation and is an important risk factor for cellular rejection. While CMV causes damage to native organs through direct viral cytopathic effect, CMV-mediated cardiac allograft rejection is attributed to “immune dysregulation,” the mechanisms for which are not well understood. This information is critical to the field of transplantation, as treatment of cellular rejection, even in cases with concurrent CMV viremia, includes immunosuppression, an approach that confers risk of abrogation of viremic control and precipitation of CMV-related morbidity, including worsening rejection and other opportunistic infections. While cellular rejection is mediated by cytotoxic T lymphocytes (CTL), recent paradigm-shifting work has demonstrated that activated mononuclear-phagocytic (MP) cells also have a critical role in driving allograft rejection through trained immunity. These findings are intriguing in the context of CMV-mediated graft rejection as MP cells are an important CMV viral reservoir. The central hypothesis being tested in this proposal is that CMV-mediated dysregulation of the MP system triggers a unique rejection phenotype that drives the infiltration of distinctly proliferative and alloreactive CTL into cardiac allograft tissue. The objective of this proposal is to leverage multi-omics strategies, including scRNA-seq and CITE-seq, to decode the allograft tissue microenvironment in rejection during CMV viremia, with a focus on understanding how MP cells orchestrate alloreactive CTL responses (Aim 1) and determine if tissue MP and CTL cell states are reflected in the blood (Aim 2). To accomplish this, we will study endomyocardial tissue and paired blood from a cohort of 200 heart transplant recipients at the Massachusetts General Hospital (MGH) Transplant Center. These studies will distinguish CMV-mediated and CMV-independent mechanisms of cellular rejection, allowing for the development of diagnostic tools and rejection treatment strategies tailored to viral or non-viral causes, promoting precision medicine in cardiac transplantation. Dr. Sen will perform the work in this K08 proposal in the Center for Immunology and Inflammatory diseases (CIID) at MGH under the mentorship of Dr. Andrew Luster and Dr. Alexandra-Chloe Villani. The CIID is a state-of-the-art multidisciplinary research center focused on mechanisms of immune-mediated inflammatory diseases and is the foundation for immunology research at MGH. Dr. Sen has devised a career development plan consisting of coursework and hands-on training in single-cell ‘omics,’ bioinformatics, the biology of alloimmunity and tolerance, cardiac tissue microenvironments, in vivo host-pathogen interactions, and human subjects research, as well as organized a Training Advisory Committee, chaired by Dr. Joren Madsen, to provide expertise and assistance in these areas. The K08 award will provide Dr. Sen with the intellectual and technical training necessary to become an independent, R01-funded investigator with expertise in host-pathogen interactions and immune dysregulation in transplantation.

项目摘要 人类巨细胞病毒（CMV）是使心脏移植复杂的最常见病毒感染，IS 细胞排斥的重要危险因素。而CMV通过直接病毒造成了对本地器官的损害 CMV介导的心脏同种异体移植排斥反应归因于“免疫失调”， 其机制尚不清楚。此信息对移植领域至关重要， 即使在同时发生CMV病毒血症的情况下，细胞排斥的治疗也包括免疫抑制 承认废除病毒控制风险和与CMV相关的发病率的危险的方法，包括 拒绝和其他机会感染的恶化。细胞排斥是由细胞毒性T介导的 淋巴细胞（CTL），最近的范式转移工作表明活化的单核斑点细胞 （MP）细胞在通过受过训练的免疫力驱动同种异体移植排斥中也具有关键作用。这些发现是 在CMV介导的移植物排斥反应的背景下，作为MP细胞是重要的CMV病毒储存剂。这 该提案中测试的中心假设是CMV介导的MP系统失调触发A 独特的拒绝表型将明显增殖和同种异体CTL渗入心脏 同种异体移植组织。该提案的目的是利用包括scrna-seq和包括 引用seq，解释在CMV病毒血症期间排斥反应中的同种异体组织微环境，重点 了解MP细胞如何协调同种反应性的CTL响应（AIM 1），并确定组织MP和 CTL细胞状态反映在血液中（AIM 2）。为此，我们将研究心肌组织和 来自马萨诸塞州综合医院（MGH）的200名心脏移植接受者的队列配对的血液 移植中心。这些研究将区分CMV介导的细胞无关和CMV独立的机制 拒绝，允许开发诊断工具和拒绝治疗策略，该策略是根据病毒或 非病毒原因，促进心脏移植中的精确药物。森博士将在此工作 在MGH的免疫学和炎症性疾病中心（CIID）的K08提案。 Andrew Luster博士和Alexandra-Chloe Villani博士。 CIID是一项最先进的多学科研究 专注于免疫介导的炎症性疾病机制，是 MGH的免疫学研究。森博士制定了一项职业发展计划，包括课程和 单细胞“ Omics”，生物信息学，同种免疫和耐受性的生物学，心脏组织的动手培训 微环境，体内宿主 - 病原体相互作用和人类受试者研究，并有组织 培训咨询委员会由Joren Madsen博士主持，以在这些领域提供专业知识和帮助。 K08奖将为SEN博士提供成为一项必要的知识和技术培训 独立的R01资助的研究者在宿主 - 病原体相互作用和免疫失调方面具有专业知识 移植。