Decoding synovial CD4+ T cell antigen specificities in Rheumatoid Arthritis

解码类风湿性关节炎滑膜 CD4 T 细胞抗原特异性

基本信息

批准号：
10569878
负责人：
Ayano Christine Kohlgruber
金额：
$ 9.4万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-05-01 至 2025-05-01
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10569878
关键词：
Affect Affinity Alleles Antibodies Antibody Formation Antibody Repertoire Antigens Area Arginine Arthritis Autoantibodies Autoantigens Autoimmune Diseases Award B-Cell Activation B-Lymphocytes Bar Codes Binding Bioinformatics Biological Assay CD4 Positive T Lymphocytes Cell Separation Cells Cellular biology Chronic Citrulline Clonal Expansion Clone Cells Coculture Techniques Collaborations Data Data Set Dedications Development Plans Disease Engineering Ensure Enzyme-Linked Immunosorbent Assay Enzymes Epitopes Faculty Flow Cytometry Frequencies Genetic Genetic Polymorphism Genetic Screening Genetic Transcription Goals HLA-DRB1 Helper-Inducer T-Lymphocyte Histocompatibility Antigens Class II Hospitals Human Immune Tolerance Immune response Immunologics Immunology Individual Inflammatory Joints Knowledge Laboratories Learning Libraries Link Lymphocyte Lymphocyte Biology Lymphoid Major Histocompatibility Complex Maps Mediator Mentors Molecular Molecular Target Pathogenicity Pathologic Pathology Patients Peptides Peripheral Phage ImmunoPrecipitation Sequencing Play Population Positioning Attribute Post-Translational Protein Processing Process Production Proteins RNA Research Resources Rheumatism Rheumatoid Arthritis Rheumatology Role Scientist Serum Site Specificity Susceptibility Gene Synovial Fluid Synovial Membrane T cell receptor repertoire sequencing T-Cell Antigen Receptor Specificity T-Cell Receptor T-Lymphocyte Technology Testing Tissues Training Woman Work antigen-specific T cells autoreactivity career career development citrullinated protein experience experimental study functional genomics genome-wide interest joint destruction medical schools memory CD4 T lymphocyte new therapeutic target novel pathogenic autoantibodies peripheral blood response screening seropositive technology development tenure track transcriptomics translational immunology

项目摘要

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.

项目摘要/摘要类风湿关节炎（RA）是一种普遍的自身免疫性疾病，其特征是慢性炎症导致关节破坏的过程。在免疫学上，自动反应性CD4+ T细胞在疾病病理并激活B细胞，导致滑膜内病理淋巴聚集体自身抗体产生。尽管识别通过致病性CD4+ T细胞识别的分子靶标的是了解RA的分子基础的关键第一步，我们仍然不知道绝大多数滑膜CD4+ T细胞以及这种反应率与自身抗体反应之间的关系。我们有开发了RA中CD4+ T细胞抗原发现的管道，该管道依赖于新的基于细胞的遗传筛查可以在基因组规模上映射TCR规范的技术。根据我们的初步单细胞转录组数据，我们已经确定了流体流体中的几个有趣的CD4+ T细胞群克隆扩展并开始发现其TCR目标。该建议是一项五年的研究和培训计划，科学重点是识别抗原靶标从RA滑膜中克隆膨胀的CD4+ T细胞的，并了解此类抗原与自身抗体反应。我们建议在AIM 1中绘制抗原表位并评估相应的抗原表位通过执行肽范围的抗原发现，克隆膨胀的滑膜CD4+ TCR的HLA限制屏幕。 AIM 2通过询问抗体库来剖析Artritic关节中T细胞-B细胞的协作结合规格和执行CD4+ T细胞-B细胞共培养分析。终于在AIM 3中，我们将设计抗原发现平台，使我们能够发现滑膜TCR反应性的能力瓜氨酸肽抗原，一种在RA中观察到的突出的翻译后修饰。这项研究将尖端的遗传和转录组技术与机械工作结合在一起评估RA的抗原特异性景观。它将为候选人提供一些科学的新培训购买翻译免疫学研究的领域。候选人的直接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测定法。候选人和导师Stephen Elledge博士都描述了一项特定的职业发展计划。功能基因组学和技术开发专家，以及迈克尔·布伦纳（Michael Brenner）博士，专家淋巴细胞生物学和RA，利用Brigham和妇女提供的强大资源医院和哈佛医学院。候选人的长期职业目标是获得终身教师职位领导着一组多样化的合作科学家，致力于研究抗原特定的免疫风湿和自身免疫性疾病的反应及其在治疗中的潜在应用。