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: 10507695
• 负责人: Lin Shen
• 金额: $ 19.77万
• 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: 2022-06-19 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10507695
• 关键词: Agonist; Alleles; Antigens; Autoimmune; Autoimmune Diseases; Autoimmunity; Biochemical; Cells; Chronic Disease; Coupled; Cytoplasmic Protein; Data; Defect; Dependence; Development; Disease; Environment; Equilibrium; Exhibits; Gene Expression; Gene Expression Profiling; Goals; Heterogeneity; Human; Immune; Immune Tolerance; Immune checkpoint inhibitor; Immune response; Lipids; Lymphocyte; Mentors; Morbidity - disease rate; Mus; Mutant Strains Mice; Mutation; Pathogenesis; Pathway interactions; Peptides; Peripheral; Phenotype; Physicians; Play; Preparation; Protein Tyrosine Kinase; Protein Tyrosine Phosphatase; Proteins; Publishing; Regulation; Regulatory Pathway; Regulatory T-Lymphocyte; Research; Resources; Role; Scientist; Series; Shapes; Signal Pathway; Signal Transduction; Syndrome; T cell response; T-Lymphocyte; Techniques; Testing; Thymus Gland; Training; Training Programs; Transgenic Organisms; Up-Regulation; ZAP-70 Gene; anergy; anti-PD1 antibodies; autoimmune pathogenesis; autoreactive T cell; autoreactivity; cancer immunotherapy; career; career development; design; experimental study; immune checkpoint blockade; immunoregulation; improved; insight; mortality; mouse model; mutant; novel therapeutic intervention; peripheral tolerance; prevent; programs; receptor; response; single-cell RNA sequencing; tool; transcriptomics; ubiquitin ligase

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.

项目概要/摘要 这个为期5年的K08培训计划旨在促进我（沉林博士）在以下领域的职业发展： 为我作为一名医师科学家的独立研究生涯做准备。我的长期目标是研究 自身免疫性疾病的发病机制和潜在的治疗。该提案的重点是如何抑制 通路抑制异常的 TCR 信号强度以实现外周耐受。中央指导和 培训环境将由我的主要导师 Arthur Weiss 博士提供，他是 TCR 信号传导方面的专家。 淋巴细胞信号传导、免疫等方面强大的专家顾问小组将提供进一步的专业知识 耐受性、自身免疫和转录组分析。包含实验研究、教学法和 举办研讨会是为了推动我实现职业目标。广泛的资源和支持 在加州大学旧金山分校的学习将促进我走向独立和实现长期目标。 长期以来，TCR 信号传导失调被认为在疾病的发病机制中发挥着重要作用。 自身免疫性疾病。然而，增强的 TCR 信号强度如何参与抑制途径 调节外周耐受性和预防自身免疫仍不完全清楚。细胞质的 酪氨酸激酶 ZAP70 在 TCR 信号传导中发挥着重要作用。两个ZAP70超态突变体，越强 W131A 和较弱的 R360P 会干扰自抑制并对 TCR 信号传导产生不同的影响。作为一个 由于基础信号传导增强，OTII-TCR 转基因 W131A T 细胞表现出显着上调 抑制性受体和无能表型的获得。我的ZAP70 R360P小鼠突变体 产生源于家族性严重自身免疫综合征。 R360P小鼠未能显性发育 C57BL/6 背景下的自身免疫性疾病，但表现出调节性 T 细胞和无反应性 T 细胞的扩增。 引人注目的是，OTI-TCR 转基因 R360P T 细胞表现出对弱肽和自身肽的增强反应。 Cbl-b（TCR 信号传导负调节因子）的引入缺陷导致 R360P-OTI T 细胞增强 对弱激动剂肽的反应并逆转 W131A-OTII T 细胞的功能性无反应性。因此，我 假设 TCR 信号强度的增加使 T 细胞受到抑制性的更大控制 途径，但可能赋予它们对这些抑制途径破坏的更大敏感性。在这个 建议，我将利用 R360P 和 W131A 小鼠模型来研究如何增强 TCR 信号传导 强度和抑制途径耦合在一起以维持或破坏外周耐受性。瞄准 1、使用一系列生化技术和转录组分析，我将剖析TCR信号传导是如何进行的 强度与抑制程序动态耦合。在目标 2 中，我将研究是否增强 TCR 信号传导 强度提高了对负面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制见解 并调节外周耐受性和免疫反应的质量。