Tuning peptide specifities for T cell tolerance in Type 1 diabetes

调整 1 型糖尿病 T 细胞耐受性的肽特异性

• 批准号: 10503923
• 负责人: Mark S Anderson
• 金额: $ 44.95万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10503923
• 关键词: Ablation; Address; Adoptive Transfer; Affect; Affinity; Agonist; Amino Acids; Antibodies; Antigens; Autoantigens; Autoimmune Diseases; Autoimmunity; Bacterial Artificial Chromosomes; Binding; Biochemical Pathway; Blood Glucose; CD4 Positive T Lymphocytes; Cells; Characteristics; Clonal Deletion; Collaborations; Coupling; Data; Development; Diabetes Mellitus; Diabetes prevention; Disease; Disease Outcome; Epitopes; Future; Generations; Genes; Histocompatibility Antigens Class II; Immune Tolerance; Immunotherapy; Inbred NOD Mice; Insulin; Insulin-Dependent Diabetes Mellitus; Knock-in; Knowledge; Laboratories; Lead; MHC Class II Genes; Mediating; Modeling; Mus; Mutation; Pathogenicity; Peptide/MHC Complex; Peptides; Peripheral; Positioning Attribute; Prevention; Regulatory T-Lymphocyte; Role; Specificity; T-Cell Immunologic Specificity; T-Lymphocyte; T-cell receptor repertoire; TNFSF11 gene; Testing; Therapeutic; Therapeutic Intervention; Thymus Gland; Time; Transgenic Mice; Transgenic Model; Transgenic Organisms; Translations; Variant; Work; anergy; autoreactive T cell; autoreactivity; central tolerance; diabetes pathogenesis; effector T cell; experience; experimental study; genetic association; genome wide association study; immunoregulation; improved; insight; insulitis; invariant chain; mouse model; novel; peripheral tolerance; prevent; promoter; single cell sequencing; thymus transplantation; tool; Ablation; Address; Adoptive Transfer; Affect; Affinity; Agonist; Amino Acids; Antibodies; Antigens; Autoantigens; Autoimmune Diseases; Autoimmunity; Bacterial Artificial Chromosomes; Binding; Biochemical Pathway; Blood Glucose; CD4 Positive T Lymphocytes; Cells; Characteristics; Clonal Deletion; Collaborations; Coupling; Data; Development; Diabetes Mellitus; Diabetes prevention; Disease; Disease Outcome; Epitopes; Future; Generations; Genes; Histocompatibility Antigens Class II; Immune Tolerance; Immunotherapy; Inbred NOD Mice; Insulin; Insulin-Dependent Diabetes Mellitus; Knock-in; Knowledge; Laboratories; Lead; MHC Class II Genes; Mediating; Modeling; Mus; Mutation; Pathogenicity; Peptide/MHC Complex; Peptides; Peripheral; Positioning Attribute; Prevention; Regulatory T-Lymphocyte; Role; Specificity; T-Cell Immunologic Specificity; T-Lymphocyte; T-cell receptor repertoire; TNFSF11 gene; Testing; Therapeutic; Therapeutic Intervention; Thymus Gland; Time; Transgenic Mice; Transgenic Model; Transgenic Organisms; Translations; Variant; Work; anergy; autoreactive T cell; autoreactivity; central tolerance; diabetes pathogenesis; effector T cell; experience; experimental study; genetic association; genome wide association study; immunoregulation; improved; insight; insulitis; invariant chain; mouse model; novel; peripheral tolerance; prevent; promoter; single cell sequencing; thymus transplantation; tool

Project Summary/Abstract Type 1 Diabetes (T1D) is a classical T-cell mediated autoimmune disease and substantial data implicates insulin as a dominant autoantigen in T1D disease. In the NOD mouse model of T1D, notable studies have shown that mice lacking native insulin expression, but with an altered insulin sequence to maintain blood glucose levels, are completely protected from insulitis and diabetes. Growing evidence also indicates that insulin peptide binding and orientation within MHC Class II (peptide register) is important in determining the strength of interaction and recognition by autoreactive T cells. In collaboration with the Kappler lab, we have uncovered an unusual peptide binding characteristic of the dominant insulin epitope InsB:9-23. The majority of InsB:9-23-specific CD4+ T cells in the periphery recognize insulin bound in this unusual register 3, and by knocking in a single amino acid variation (R to E) into just one copy of the insulin gene in NOD mice (Ins2EE/+), the mice are completely protected against diabetes. The development of a “super agonist” version of the insulin dominant epitopes allows us to address several key questions surrounding the biochemical pathways of peptide generation, presentation by MHC molecules, and recognitions by auto-reactive pathogenic T cells. Do mutations of the major epitope in the insulin gene allow CD4+ T effectors or Treg cells specific for these alternative epitopes to develop? Are mimotopes of these pathogenic epitopes capable of dramatically altering disease outcomes? Are we able to fine tune these epitopes to alter tolerance mechanisms to shift from deletion to Treg induction? Recent work in our lab has focused on the identification of the insulin-specific repertoire on key mouse backgrounds, and we plan to utilize these tools and well-characterized mouse models to examine the effects of altering insulin expression, thymically and extrathymically. These tools as well as our experience with the generation of numerous TCR-transgenic mouse lines will allow us to address these questions in the context of T1D. Thus, we hypothesize that alterations to epitope presentation and TCR affinity drive the tunning of the TCR repertoire towards tolerance and away from self-reactivity. Using Insulin as a model antigen, we propose to test our hypothesis through the following specific aims: Aim 1: Define the role of central tolerance upon the deletion of insulin-reactive clones Aim 2: Characterize the effects of peripheral tolerance on insulin-reactive T cells Aim 3: Explore mechanisms of dominate tolerance to understand the potential for translation into therapeutic treatments for T1D Through these experiments, we hope to gain a nuanced understanding of how changes in insulin epitopes and antigenicity drive the pathogenesis of diabetes and identify targets for future immune modulation and therapeutic intervention for T1D treatment and prevention.

项目摘要/摘要 1型糖尿病（T1D）是经典的T细胞介导的自身免疫性疾病，大量数据牵涉 胰岛素是T1D疾病中主要的自身抗原。在T1D的点头小鼠模型中，著名的研究表明 那些缺乏天然胰岛素表达的小鼠，但胰岛素序列改变以维持血糖水平， 完全保护胰岛素和糖尿病。越来越多的证据也表明胰岛素肽结合 MHC II类（肽寄存器）中的方向对于确定相互作用的强度和 自动反应性T细胞识别。与卡普勒实验室合作，我们发现了不寻常的肽 主导胰岛素发作的结合特征INSB：9-23。大多数INSB：9-23特异性CD4+ T细胞 在该异常寄存器3中结合的外围识别胰岛素中，并通过敲击单个氨基酸 在NOD小鼠（INS2EE/+）中，只有一个胰岛素基因的变异（r至e），完全保护小鼠 针对糖尿病。 胰岛素主要表位的“超级激动剂”版本的开发使我们能够解决几个 围绕肽产生的生化途径的关键问题，MHC分子呈现， 自身反应性致病细胞的识别。胰岛素基因中主要表位的突变允许 CD4+ T效应子或Treg细胞特异性对于这些替代表位的发展？是这些的模仿 能够大大改变疾病结局的致病表位？我们可以微调这些表位吗 改变耐受性机制以从缺失转向Treg诱导？我们实验室的最新工作重点是 在关键鼠标背景上识别胰岛素特定曲目，我们计划使用这些工具 以及特征良好的小鼠模型，以检查胰岛素表达的效果， 谨慎。这些工具以及我们与大量TCR-转基因鼠标的经验 线条将使我们能够在T1D的背景下解决这些问题。 这就是我们假设对情节演示和TCR亲和力的改变驱动了隧道 TCR曲目对宽容和远离自我反应。使用胰岛素作为模型抗原，我们 通过以下特定目的来检验我们的假设的建议： 目标1：定义中心耐受性在删除胰岛素反应克隆中的作用 AIM 2：表征周围耐受性对胰岛素反应T细胞的影响 AIM 3：探索占主导地位的机制，以了解转化为翻译的潜力 T1D的治疗治疗 通过这些实验，我们希望对胰岛素表位的变化以及 抗原性驱动糖尿病的发病机理，并确定未来免疫调节的靶标 T1D治疗和预防的治疗干预。