Dissecting the interactions between TCR signaling strength and inhibitory pathways

剖析 TCR 信号强度和抑制途径之间的相互作用

• 批准号: 10895664
• 负责人: Lin Shen
• 金额: $ 15.36万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-19 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10895664
• 关键词: Dissecting; interactions; between; TCR; signaling

PROJECT SUMMARY/ABSTRACT This 5-year K08 training program is designed to facilitate my (Dr. Lin Shen) career development in preparation of my independent research career as a physician-scientist. My long-term goal is to study the pathogenesis and, potentially, the therapy of autoimmune diseases. This proposal focuses on how inhibitory pathways restrain abnormal TCR signaling strength to achieve peripheral tolerance. Central guidance and the training environment will be provided by my primary mentor, Dr. Arthur Weiss, an expert in TCR signaling. Further expertise will be provided by a strong advisory panel of experts in lymphocyte signaling, immune tolerance, autoimmunity and transcriptomic profiling. A training plan with experimental research, didactics, and seminars has been developed to advance me towards my career goals. The extensive resources and support at the UCSF will facilitate my pathway to independence and my long-term goals. Dysregulation of TCR signaling has long been recognized to play important roles in the pathogenesis of autoimmune diseases. However, how inhibitory pathways are engaged by enhanced TCR signaling strength to regulate peripheral tolerance and prevent autoimmunity remains incompletely understood. The cytoplasmic tyrosine kinase ZAP70 plays a requisite role in TCR signaling. Two ZAP70 hypermorphic mutants, the stronger W131A and the weaker R360P, interfere with autoinhibition and have varying effects on TCR signaling. As a consequence of enhanced basal signaling, OTII-TCR transgenic W131A T cells exhibit marked upregulation of inhibitory receptors and acquisition of an anergic phenotype. The ZAP70 R360P mouse mutant that I generated derives from a familial severe autoimmune syndrome. R360P mice failed to develop overt autoimmune disease on a C57BL/6 background but exhibited expansion of regulatory and anergic T cells. Strikingly, OTI-TCR transgenic R360P T cells showed enhanced responses to weak and self-peptides. Introducing deficiency in Cbl-b, a negative regulator for TCR signaling, resulted in enhanced R360P-OTI T cell responses to a weak agonist peptide and reversed functional anergy in W131A-OTII T cells. Therefore, I hypothesize that increased TCR signaling strength renders T cells subject to greater control by inhibitory pathways but may confer upon them greater sensitivity to disruption of these inhibitory pathways. In this proposal, I will take advantage of the R360P and W131A mouse models to study how enhanced TCR signaling strength and inhibitory pathways are coupled together to either maintain or subvert peripheral tolerance. In Aim 1, using a range of biochemical techniques and transcriptomic profiling, I will dissect how TCR signaling strength is dynamically coupled to inhibitory programs. In Aim 2, I will study whether augmented TCR signaling strength promotes sensitivity to the disruption of negative regulatory pathways. This study will provide new mechanistic insights into how these opposing signaling pathways integrate to impact T cell antigen sensitivity and to regulate peripheral tolerance and the quality of immune responsiveness.

项目摘要/摘要 这项为期5年的K08培训计划旨在促进我（Lin Shen博士）职业发展 作为医师科学家的独立研究生涯的准备。我的长期目标是研究 发病机理，可能是自身免疫性疾病的治疗。该提案重点是抑制性 途径限制异常TCR信号传导强度，以达到外围耐受性。中央指导和 TCR信号专家Arthur Weiss博士将提供培训环境。 强大的淋巴细胞信号专家咨询小组将提供进一步的专业知识，免疫 公差，自身免疫和转录组分析。实验研究，教学和 研讨会是为了促进我的职业目标发展的。广泛的资源和支持 在UCSF，将促进我的独立途径和长期目标。 长期以来，TCR信号的失调一直被认为在 自身免疫性疾病。但是，通过增强的TCR信号强度使抑制途径如何参与 调节外围耐受性并预防自身免疫性仍然不完全理解。细胞质 酪氨酸激酶ZAP70在TCR信号传导中起必要的作用。两个ZAP70超态突变体，较强 W131a和较弱的R360p，会干扰自身抑制，并对TCR信号传导产生不同的影响。作为 增强基础信号传导的结果，OTII-TCR转基因W131A T细胞表现出标志性上调 抑制受体和厌氧表型的获取。 zap70 r360p小鼠突变体 产生的源自家族严重的自身免疫性综合征。 R360p小鼠未能开发公开 在C57BL/6背景下进行自身免疫性疾病，但表现出调节和厌食性T细胞的扩展。 令人惊讶的是，OTI-TCR转基因R360p T细胞显示出对弱和自肽的反应增强。 引入CBL-B缺乏（一种用于TCR信号传导的负调节剂）导致R360p-Oti T细胞增强 对W131a-Otii T细胞中的弱激动剂肽的反应和逆转功能性反向。因此，我 假设提高TCR信号强度会使T细胞受到抑制更大的控制 途径但可能会赋予他们对这些抑制途径的破坏更大的敏感性。在这个 提案，我将利用R360p和W131a鼠标模型来研究增强TCR信号的方式 强度和抑制途径耦合在一起，以维持或颠覆外围耐受性。目标 1，使用一系列的生化技术和转录组分析，我将剖析TCR信号传导 强度与抑制程序动态耦合。在AIM 2中，我将研究是否增强TCR信号传导 力量促进对负调节途径破坏的敏感性。这项研究将提供新的 关于这些相反的信号通路如何整合以影响T细胞抗原敏感性的机械洞察力 并调节外周耐受性和免疫反应性的质量。