Identification of the cognate epitopes of autoreactive T cells in Type 1 Diabetes

1 型糖尿病自身反应性 T 细胞同源表位的鉴定

• 批准号: 10264075
• 负责人: Alok joglekar
• 金额: $ 15.91万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-14 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10264075
• 关键词: Affect; Alleles; American; Antigens; Autoantigens; Autoimmune; Binding; Biological Assay; CD4 Antigens; CD4 Positive T Lymphocytes; CD8B1 gene; Cells; Clone Cells; Data Set; Disease; Epitopes; FOXP3 gene; Foundations; Future; Gene Expression; Genes; Goals; Histocompatibility Antigens Class II; Human; Immune Tolerance; Immune response; Immune system; Immunotherapy; In Vitro; Inbred NOD Mice; Individual; Insulin; Insulin-Dependent Diabetes Mellitus; Islets of Langerhans; Jurkat Cells; Knowledge; Libraries; Major Histocompatibility Complex; Maps; Mass Spectrum Analysis; Mediating; Methods; Mus; Pancreas; Pathogenesis; Patients; Peptide/MHC Complex; Peptides; Protocols documentation; Publishing; Regulatory T-Lymphocyte; Reporter; Research; Sampling; Signal Transduction; Sorting - Cell Movement; Source; Specificity; T cell receptor repertoire sequencing; T-Cell Antigen Receptor Specificity; T-Cell Receptor; T-Lymphocyte; T-Lymphocyte Epitopes; T-cell receptor repertoire; Techniques; Technology; Tissues; arm; autoreactive T cell; autoreactivity; base; cell killing; central tolerance; combat; cytotoxicity; design; diabetes pathogenesis; diabetogenic; effector T cell; in vitro Assay; islet; mouse model; novel; novel therapeutic intervention; peripheral tolerance; prevent; programs; receptor; single cell sequencing; targeted treatment

ABSTRACT Type 1 Diabetes (T1D), affects approximately 4 million individuals worldwide, including 1.6 million Americans. T1D is caused by progressive destruction of pancreatic  cells, resulting in a significantly diminished capacity to produce insulin. Diabetogenic CD8+ and CD4+ effector T cells that infiltrate pancreatic islets mediate  cell destruction in an antigen-specific manner by recognizing peptide epitopes presented on class I and class II MHC molecules respectively. In contrast, regulatory T cells can suppress diabetogenic T cells, thereby preventing T1D pathogenesis. Recognition of epitopes presented by  cells is critical for the function of these autoreactive T cells. Only a small number of self-epitopes recognized by autoreactive CD8+, CD4+ effector and regulatory T cells in T1D have been uncovered. However, the epitopes recognized by the majority of islet-infiltrating T cells are not known. The knowledge of these epitopes is critical for understanding disease pathogenesis and for developing targeted therapies. Currently, widely applicable and efficient methods for T cell antigen discovery for uncovering autoreactivity are lacking. The overarching goal of this project is to uncover the cognate epitopes of autoreactive T cells in T1D using a novel and generalizable antigen discovery technology developed by our group. In this proposal, we will employ T cell epitope discovery using Signaling and Antigen-presenting Bifunctional Receptors (SABRs) to identify the epitopes recognized by effector and regulatory T cells in a mouse model of T1D, NOD mice. We propose that constructing a library of epitopes derived from genes expressed specifically in pancreatic  cells will lead to identification of novel targets of islet-infiltrating T cells. We will construct epitope libraries from published mass spectrometry and gene expression datasets from NOD mice. We will obtain islet-reactive TCRs by performing single cell TCR sequencing on pancreatic islets of NOD mice. Using  cell derived SABR libraries, we will determine the cognate epitopes of islet-reactive TCRs and validate them in vitro. The epitopes identified by these studies will lead to future studies aiming to understand the breakage of immune tolerance by autoreactive T cells and to develop targeted immunotherapy approaches to combat T1D. This approach will also establish the foundation for antigen discovery for T cells from T1D patient samples in affiliation with Human Islet Research Network. 1

摘要 1型糖尿病（T1D）影响全球约400万人，其中包括160万美国人。 T1D是由胰腺炎细胞的进行性破坏引起的，导致胰腺炎细胞的增殖能力显著降低。 产生胰岛素。浸润胰岛的致糖尿病性CD8+和CD4+效应T细胞介导胰岛细胞增殖 通过识别I类和II类MHC上呈现的肽表位以抗原特异性方式进行破坏 分子分别。相反，调节性T细胞可以抑制致糖尿病性T细胞，从而预防T1D 发病机制识别由T细胞呈递的表位对于这些自身反应性T细胞的功能至关重要。 细胞只有少数自身抗原表位被自身反应性CD8+、CD4+效应子和调节性T细胞识别， T1D的细胞已经被发现。然而，大多数胰岛浸润性T细胞识别的表位 不知道。这些表位的知识对于理解疾病的发病机制和预防疾病是至关重要的。 开发靶向治疗。目前，用于T细胞抗原发现的广泛适用且有效的方法， 缺乏对自身反应性的揭示该项目的总体目标是揭示的同源表位的 使用我们开发的一种新的和可推广的抗原发现技术， 组在这个提议中，我们将使用信号传导和抗原呈递来发现T细胞表位。 双功能受体（SABR），用于识别小鼠中效应和调节T细胞识别的表位 T1D、NOD小鼠模型。我们建议构建一个来自表达基因的表位库， 特别是在胰腺癌细胞中的研究将导致对胰岛浸润性T细胞的新靶点的鉴定。我们将 从公开的质谱和来自NOD小鼠的基因表达数据集构建表位文库。 我们将通过对NOD小鼠的胰岛进行单细胞TCR测序来获得胰岛反应性TCR。 使用胰岛细胞来源的SABR文库，我们将确定胰岛反应性TCR的同源表位，并验证 在vitro。这些研究所确定的表位将导致未来的研究，旨在了解 自身反应性T细胞破坏免疫耐受，并开发靶向免疫治疗方法， 战斗T1D。该方法也将为T1D患者T细胞的抗原发现奠定基础 人类胰岛研究网络（Human Islet Research Network） 1