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.

项目概要 人类巨细胞病毒 (CMV) 是导致心脏移植复杂化的最常见病毒感染， 细胞排斥的重要危险因素，而 CMV 通过直接病毒对天然器官造成损害。 细胞病变效应，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，因为该信息对于移植领域至关重要。 细胞排斥的治疗，即使是在并发 CMV 病毒血症的情况下，也包括免疫抑制、 导致病毒血症控制失效和巨细胞病毒相关发病率下降的风险的方法，包括 排斥反应恶化和其他机会性感染，而细胞排斥反应是由细胞毒性 T 介导的。 淋巴细胞（CTL），最近的范式转换工作表明，激活的单核吞噬细胞 (MP) 细胞在通过训练有素的免疫驱动同种异体移植排斥方面也发挥着关键作用。 由于 MP 细胞是重要的 CMV 病毒库，因此在 CMV 介导的移植排斥反应中这一点很有趣。 本提案中测试的中心假设是 CMV 介导的 MP 系统失调会触发 独特的排斥表型，驱动明显增殖和同种异体反应性的 CTL 渗入心脏 该提案的目标是利用多组学策略，包括 scRNA-seq 和 CITE-seq，解码 CMV 病毒血症期间排斥反应中的同种异体移植组织微环境，重点关注 了解 MP 细胞如何协调同种反应性 CTL 反应（目标 1）并确定组织 MP 和 CTL 细胞状态反映在血液中（目标 2）。为了实现这一目标，我们将研究心内膜心肌组织和 来自麻省总医院 (MGH) 200 名心脏移植接受者的配对血液 移植中心。这些研究将区分 CMV 介导的和 CMV 独立的细胞机制。 排斥反应，允许开发针对病毒或病毒量身定制的诊断工具和排斥反应治疗策略 Sen 博士将致力于非病毒原因，促进心脏移植领域的精准医疗。 在麻省总医院免疫学和炎症疾病中心 (CIID) 的指导下提出 K08 提案 Andrew Luster 博士和 Alexandra-Chloe Villani 博士是一项最先进的多学科研究。 该中心专注于免疫介导的炎症性疾病的机制，是 Sen 博士在 MGH 进行免疫学研究，制定了一个职业发展计划，包括课程作业和 单细胞“组学”、生物信息学、同种免疫和耐受生物学、心脏组织的实践培训 微环境、体内宿主-病原体相互作用和人类受试者研究，并组织了一次 培训咨询委员会由 Joren Madsen 博士担任主席，负责在这些领域提供专业知识和帮助。 K08 奖将为 Sen 博士提供成为一名 由 R01 资助的独立研究者，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。