High-throughput identification and transcriptional analysis of autoreactive T cells in individuals with membranous nephropathy.

膜性肾病患者自身反应性 T 细胞的高通量鉴定和转录分析。

基本信息

批准号：
10725558
负责人：
Paolo Cravedi
金额：
$ 26.9万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
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：
2023-08-16 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10725558
关键词：
Adult Affect Aftercare Alloantigen Antibodies Antigen Targeting Antigens Autoantibodies Autoimmune Autoimmune Diseases Autologous B-Lymphocytes Biological Biological Markers Blood Volume Blood specimen CD4 Positive T Lymphocytes Cell Compartmentation Cells Characteristics Circulation Clinical Clinical Research Cloning Complement Couples DNA Sequence Data Deposition Development Diagnosis Disease Disease remission End stage renal failure Epitope spreading Epitopes Exposure to Fc Receptor Functional disorder Gene Expression Profile Gene Library Generations Genetic Transcription Human Human Herpesvirus 4 Immune Tolerance Immune response Immunologic Monitoring In Vitro Individual Inflammatory Injury Libraries Link Maps Mediating Membranous Glomerulonephritis Molecular Nephrotic Syndrome Outcome Pathogenicity Pathway interactions Patients Peptide Receptor Peptides Peripheral Blood Mononuclear Cell Phospholipase Phospholipase A2 Production Receptor Cell Recombinants Renal glomerular disease Research Research Design Role Sampling Severity of illness Signal Transduction Specificity T cell regulation T cell response T-Cell Immunologic Specificity T-Cell Receptor T-Cell Receptor Genes T-Lymphocyte T-cell receptor repertoire Techniques Technology Testing Therapeutic Therapeutic Uses Tumor Antigens United States National Institutes of Health Up-Regulation active method alpha-beta T-Cell Receptor autoreactive T cell cost effective design experience glomerular basement membrane high reward high risk innovation innovative technologies kidney biopsy novel strategies pathogen patient screening patient subsets podocyte prevent prophylactic receptor rituximab screening single-cell RNA sequencing therapeutic target tool

Adult Affect Aftercare Alloantigen Antibodies Antigen Targeting Antigens Autoantibodies Autoimmune Autoimmune Diseases Autologous B-Lymphocytes Biological Biological Markers Blood Volume Blood specimen CD4 Positive T Lymphocytes Cell Compartmentation Cells Characteristics Circulation Clinical Clinical Research Cloning Complement Couples DNA Sequence Data Deposition Development Diagnosis Disease Disease remission End stage renal failure Epitope spreading Epitopes Exposure to Fc Receptor Functional disorder Gene Expression Profile Gene Library Generations Genetic Transcription Human Human Herpesvirus 4 Immune Tolerance Immune response Immunologic Monitoring In Vitro Individual Inflammatory Injury Libraries Link Maps Mediating Membranous Glomerulonephritis Molecular Nephrotic Syndrome Outcome Pathogenicity Pathway interactions Patients Peptide Receptor Peptides Peripheral Blood Mononuclear Cell Phospholipase Phospholipase A2 Production Receptor Cell Recombinants Renal glomerular disease Research Research Design Role Sampling Severity of illness Signal Transduction Specificity T cell regulation T cell response T-Cell Immunologic Specificity T-Cell Receptor T-Cell Receptor Genes T-Lymphocyte T-cell receptor repertoire Techniques Technology Testing Therapeutic Therapeutic Uses Tumor Antigens United States National Institutes of Health Up-Regulation active method alpha-beta T-Cell Receptor autoreactive T cell cost effective design experience glomerular basement membrane high reward high risk innovation innovative technologies kidney biopsy novel strategies pathogen patient screening patient subsets podocyte prevent prophylactic receptor rituximab screening single-cell RNA sequencing therapeutic target tool

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项目摘要

SUMMARY Membranous nephropathy (MN) is a glomerular disease due to the deposition of anti-podocyte antibodies in the subepithelial space of the glomerular basement membrane. This leadis to complement-mediated podocyte injury and, in ~30% of patients, to the development of end stage kidney disease within 10 years from diagnosis. Most of the research in MN has focused on the autoantibody specificities and has led to the identification of phospholipase 2A receptor (PLA2R) as the main target antigen in 70%-80% of the patients. Conversely, little is known on autoreactive T cells, despite the evidence from other antibody-mediated autoimmune diseases clearly showing a critical pathogenic role of autoreactive T cells, efficacy of T cell targeting treatments, and our preliminary data documenting that autoreactive T cells are present in the circulation of MN patients. Aim 1 of the present project will develop a new strategy to capture the T cell receptor (TCR) repertoire of PLA2R-reactive T cells in MN patients. We will apply high-throughput paired T cell receptor alpha and beta chain DNA sequence capture for renewable TCR gene library generation and functional screening from patient T cells. We will physically link TCR gene sequences for cloning into cellular display libraries, enabling repeated in vitro functional analyses of TCRs in ways that are impossible with alternative available techniques. Aim 2 will apply these technologies for the study of the single-cell transcriptional profile of CD4+ T cells. These data will reveal the characteristics of autoreactive TCRs in a previously collected samples from MN patients with remission versus active disease, setting the stage for expanded clinical studies. Overall, this project will collect and analyze immune response data from MN patients with unprecedented molecular scope and scale. Our approach is innovative because it couples carefully designed clinical sample sets with new high-throughput approaches in TCR analysis to comprehensively interrogate the molecular mechanisms of T cell responses in MN patients. The proposed research is significant because it will provide important tools for mechanistic studies aimed at identifying new biomarkers and therapeutic targets for MN patients.

概括膜性肾病（MN）是肾小球疾病，这是由于抗幼体抗体的沉积在肾小球基底膜的上皮下空间中。这是补体介导的足细胞的领导受伤，约30％的患者在诊断后的10年内发展为终阶段肾脏疾病。 MN的大多数研究都集中在自身抗体特异性上，并导致了鉴定磷脂酶2A受体（PLA2R）作为70％-80％的患者的主要靶抗原。相反，几乎没有尽管有其他抗体介导的自身免疫性疾病的证据，但在自动反应性T细胞上已知显示自动反应性T细胞的关键致病作用，T细胞靶向处理的功效，我们初步数据记录了MN患者的循环中存在自动反应性T细胞。本项目的目标1将制定一种新的策略来捕获T细胞受体（TCR）曲目 MN患者的PLA2R反应性T细胞。我们将应用高通量配对的T细胞受体α和β链可再生TCR基因文库产生的DNA序列捕获和患者T细胞的功能筛选。我们将实际将TCR基因序列连接到克隆到细胞显示库中，从而在体外重复通过替代可用技术是不可能的，对TCR的功能分析。 AIM 2将适用这些技术用于研究CD4+ T细胞的单细胞转录谱。这些数据将显示从MN患者先前收集的样品中自动反应性TCR的特征相对于活跃疾病，为扩大的临床研究奠定了基础。总体而言，该项目将收集和分析来自前所未有的MN患者的免疫反应数据分子范围和比例。我们的方法具有创新性，因为它融合了精心设计的临床样本在TCR分析中采用新的高通量方法的设置，以全面询问分子 MN患者T细胞反应的机制。拟议的研究很重要，因为它将提供机械研究的重要工具，旨在识别MN的新生物标志物和治疗靶标患者。