Decoding synovial CD4+ T cell antigen specificities in Rheumatoid Arthritis

项目总结/摘要 类风湿性关节炎（RA）是一种常见的自身免疫性疾病， 导致联合破坏的过程。在免疫学上，自身反应性CD 4 + T细胞在免疫系统中起着核心作用。 疾病病理学和激活B细胞，导致滑膜内病理性淋巴聚集， 自身抗体产生。虽然鉴定致病性CD 4 + T细胞识别的分子靶点是一个重要的研究课题， 尽管这是了解RA分子基础的关键第一步，但我们仍然不知道RA的抗原靶点。 绝大多数滑膜CD 4 + T细胞以及这些反应性如何与自身抗体应答相关。我们有 开发了一种新的基于细胞的基因筛选的RA中CD 4 + T细胞抗原发现管道 该技术能够在基因组规模上绘制TCR特异性。根据我们初步的单细胞 转录组学数据，我们已经确定了几个有趣的CD 4 + T细胞群在滑液中， 克隆扩增，并已开始发现其TCR靶点。 这项建议是一项为期五年的研究和培训计划，其科学重点是确定抗原靶点 克隆扩增的RA滑膜CD 4 + T细胞，并了解这些抗原如何与 自身抗体反应我们在目的1中提出绘制抗原表位并评估相应的免疫原性。 通过进行全肽组抗原发现对克隆扩增的滑膜CD 4 + TCR进行HLA限制 卡位目的2通过询问抗体库来剖析关节炎关节中T细胞-B细胞的协作 结合特异性和进行CD 4 + T细胞-B细胞共培养测定。最后，在目标3中，我们将设计一个 抗原发现平台，使我们能够发现滑膜TCR反应性， 瓜氨酸肽抗原，一个突出的翻译后修饰观察到RA。 这项研究将尖端的遗传和转录组技术与机械工作相结合， 评估RA的抗原特异性景观。它将为候选人提供几个科学的新培训 从事转化免疫学研究的领域。候选人的近期职业发展目标是 获得生物信息学分析、抗体分析技术和人类免疫学检测的经验。 一个具体的职业发展计划是由候选人和导师，博士斯蒂芬Elledge描述， 功能基因组学和技术开发专家，以及医学博士迈克尔布伦纳， 淋巴细胞生物学和类风湿关节炎，利用强大的资源，可在布里格姆和妇女的 医院和哈佛医学院。候选人的长期职业目标是获得终身教职 领导一个致力于研究抗原特异性免疫的不同合作科学家小组的职位 风湿性和自身免疫性疾病的反应及其潜在的治疗应用。

Project Summary/Abstract Rheumatoid arthritis (RA) is a prevalent autoimmune disorder characterized by chronic inflammatory processes that lead to joint destruction. Immunologically, autoreactive CD4+ T cells play a central role in disease pathology and activate B cells leading to pathologic lymphoid aggregates within the synovium and autoantibody production. Although identifying the molecular targets recognized by pathogenic CD4+ T cells is a critical first step in understanding the molecular basis of RA, we still do not know the antigenic targets for the vast majority of synovial CD4+ T cells and how such reactivities relate to autoantibody responses. We have developed a pipeline for CD4+ T cell antigen discovery in RA that relies on a new, cell-based genetic-screening technology that enables mapping of TCR specificities at genome scale. Based on our preliminary single-cell transcriptomic data, we have identified several interesting CD4+ T cell populations in synovial fluid that are clonally expanded and have begun to discover their TCR targets. This proposal is a five-year research and training plan with a scientific focus on identifying the antigenic targets of clonally expanded CD4+ T cells from RA synovium and understanding how such antigens relate to autoantibody responses. We propose in Aim 1 to map the antigenic epitopes and assess the corresponding HLA-restriction of clonally expanded synovial CD4+ TCRs by performing peptidome-wide antigen discovery screens. Aim 2 dissects T cell-B cell collaboration in the arthritic joint by interrogating antibody repertoire binding specificities and performing CD4+ T cell-B cell co-culture assays. Finally in Aim 3, we will engineer an antigen discovery platform to enable our ability to uncover synovial TCR reactivities against citrullinated-peptide antigens, a prominent post-translational modification observed in RA. This study combines cutting-edge genetic and transcriptomic technologies with mechanistic work to critically evaluate the antigen-specific landscape of RA. It will provide the candidate new training in several scientific areas to pursue translational immunology research. The candidate’s immediate career development goals are to gain experience with bioinformatic analysis, antibody profiling technologies, and human immunology assays. A specific career development plan is described by both the candidate and the mentors, Dr. Stephen Elledge, an expert in functional genomics and technology development, and Dr. Michael Brenner MD, an expert in lymphocyte biology and RA, taking advantage of the powerful resources available at Brigham and Women’s Hospital and Harvard Medical School. The candidate’s long-term career goal is to attain a tenure-track faculty position leading a diverse group of collaborative scientists dedicated to studying antigen specific immune responses in rheumatic and autoimmune diseases and their potential applications for therapy.