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.

项目概要/摘要类风湿性关节炎（RA）是一种常见的自身免疫性疾病，其特征是慢性炎症在导致关节破坏的免疫过程中，自身反应性 CD4+ T 细胞在其中发挥着核心作用。疾病病理学并激活 B 细胞，导致滑膜内病理性淋巴聚集，尽管识别致病性 CD4+ T 细胞识别的分子靶标是一种自身抗体的产生。了解 RA 分子基础的关键第一步，我们仍然不知道 RA 的抗原靶标绝大多数滑膜 CD4+ T 细胞以及这种反应与自身抗体反应的关系。开发了一条在 RA 中发现 CD4+ T 细胞抗原的管道，该管道依赖于一种新的基于细胞的基因筛选基于我们初步的单细胞，能够在基因组范围内绘制 TCR 特异性的技术。转录组数据，我们在滑液中鉴定出了几个有趣的 CD4+ T 细胞群，它们是克隆扩展并已开始发现其 TCR 靶点。该提案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目标从 RA 滑膜中克隆扩增的 CD4+ T 细胞，并了解这些抗原如何与我们在目标 1 中建议绘制抗原表位并评估相应的反应。通过进行全肽组抗原发现对克隆扩展的滑膜 CD4+ TCR 进行 HLA 限制目标 2 通过研究抗体库来剖析关节炎关节中的 T 细胞与 B 细胞的协作。结合特异性并进行 CD4+ 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 反应瓜氨酸肽抗原，是在 RA 中观察到的一种重要的翻译后修饰。这项研究将尖端的遗传和转录组技术与机械工作相结合，以批判性地评估 RA 的抗原特异性情况它将为候选人提供多项科学方面的新培训。候选人的近期职业发展目标是从事转化免疫学研究的领域。获得生物信息分析、抗体分析技术和人类免疫学测定方面的经验。候选人和导师 Stephen Elledge 博士描述了具体的职业发展计划，功能基因组学和技术开发专家 Michael Brenner 博士（医学博士）淋巴细胞生物学和 RA，利用布莱根妇女医院强大的资源医院和哈佛医学院的候选人的长期职业目标是获得终身教职。领导致力于研究抗原特异性免疫的多元化合作科学家小组风湿病和自身免疫性疾病的反应及其潜在的治疗应用。