Inhibition of ZAP-70 to selectively attenuate autoimmune disease-inducing T cells

抑制 ZAP-70 可选择性减弱自身免疫性疾病诱导 T 细胞

• 批准号: 8617722
• 负责人: Byron Benton Au-Yeung
• 金额: $ 12.84万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-18 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8617722
• 关键词: Achievement; Animal Model; Attenuated; Autoantigens; Autoimmune Diseases; Autoimmune Process; Autoimmunity; Award; CD4 Positive T Lymphocytes; CD8B1 gene; California; Cell physiology; Cells; Chronic; Collagen Arthritis; Data; Disease; Disease model; Disease susceptibility; Doctor of Philosophy; Environment; Experimental Autoimmune Encephalomyelitis; Exposure to; Generations; Genes; Genetic Polymorphism; Goals; HLA-DR4 Antigen; Health; IL17 gene; IL2RA gene; Immunology; Immunosuppression; Impairment; Infiltration; Inflammation; Interleukin-17; K-Series Research Career Programs; Laboratories; Learning; Mediating; Mentors; Mentorship; Microbiology; Modeling; Molecular; Multiple Sclerosis; Mus; PTPN22 gene; Pathogenesis; Pathology; Patients; Pharmaceutical Preparations; Phosphotransferases; Process; Production; Protein Tyrosine Kinase; Proteins; Receptor Signaling; Regulatory T-Lymphocyte; Relative (related person); Research; Rheumatoid Arthritis; Rodent Model; Role; San Francisco; Scientist; Signal Transduction; Signaling Molecule; Stimulus; Students; Symptoms; System; T Cell Receptor Signaling Pathway; T cell anergy; T cell differentiation; T cell response; T cell therapy; T memory cell; T-Cell Activation; T-Cell Receptor; T-Lymphocyte; T-Lymphocyte Subsets; Testing; Th1 Cells; Th2 Cells; Therapeutic; Time; Tissues; Titrations; Training; Tyrosine Kinase Inhibitor; Universities; Work; ZAP-70 Gene; anergy; autoreactive T cell; cytokine; design; effective therapy; experience; genome wide association study; in vivo; inhibitor/antagonist; insight; interest; joint destruction; mouse model; novel; novel strategies; novel therapeutics; post-doctoral training; prevent; response; rheumatologist; small molecule; technology/technique; tool; ubiquitin ligase

DESCRIPTION (provided by applicant): Candidate and Training Environment: I am seeking a K01 Mentored Career Development Award to obtain additional training, which will facilitate my transition into an independent scientist. My long-term research goal is to understand how manipulation of T cell receptor signals can be used to alter or prevent diverse T cell functions, i the interest of developing therapeutic strategies for T cell mediated diseases. As a PhD student under the mentorship of Dr. Deborah Fowell, I determined the requirements for the tyrosine kinase Itk in CD4+ T cell differentiation. During the course of those studies, I gained training in cellular T cell immunology, mostly related to the isolation, activation, differentiation, and effecor cytokine production of primary mouse CD4+ T cells. My postdoctoral training in the laboratory of Dr. Arthur Weiss has so far focused on developing and characterizing a unique experimental inhibitor for the tyrosine kinase ZAP-70. We showed that this inhibitor system is highly specific, titratable, and can rapidly and reversibly inhibit ZAP-70 kinase activity, making this system a very useful tool for modulating the magnitude of T cell receptor signaling. At this point of my postdoctoral training, the technology and techniques that I have learned to use are well suited to understand how manipulations of T cell receptor signaling can be applied to alter T cell function, which could be important for developing novel therapeutic strategies for T cell mediated autoimmune diseases, such as rheumatoid arthritis. Further training in the analysis autoimmune disease models, and the signals that drive the activation and differentiation of autoreactive T cells will allow me to develop an independent research plan and transition into an independent scientist. The proposed studies will take place under the mentorship of Dr. Arthur Weiss at the University of California, San Francisco (UCSF). Dr. Weiss is a rheumatologist, a renowned expert in the field of T cell signal transduction, and a distinguished mentor as the recipient of te UCSF Lifetime Achievement in Mentoring award. Additionally, I will be able to get advice from my collaborators/consultants at UCSF, Dr. Abul Abbas and Dr. Mark Anderson, who have considerable experience and expertise in the fields of autoimmunity and tolerance. The Department of Microbiology and Immunology at UCSF is also highly regarded and is a rich environment for gaining additional training and input for studies of autoimmunity. Research: T cell responses are associated with multiple autoimmune diseases such as rheumatoid arthritis (RA). Therefore, a therapeutic strategy for blocking T cell function could be useful for treatment of RA and other T cell-mediated autoimmune diseases. While inhibiting all T cell function would block the autoreactive T cell response, a more desirable strategy would selectively block the subset of T cells responsible for causing pathology. Previous work and preliminary data suggest that inhibition of the tyrosine kinase ZAP-70 could fit these criteria. First, the activation of conventional CD4+ and CD8+, but not regulatory T cells, requires ZAP-70 catalytic activity. Secondly, preliminary data indicates that CD4+ T cells undergoing Th17 differentiation are substantially more sensitive to ZAP-70 inhibition as compared to Th1 and Th2 differentiating cells. Together these data suggest that ZAP-70 inhibition could be very useful for treating autoimmune diseases mediated by autoreactive Th17 responses, such as RA. In Aim 1 of this proposal, I aim to determine how Th17 differentiating cells are differentially sensitive to ZAP-70 inhibition. First, I will test the hypothesis that Th17 cells are preferentially derived from cellsthat express relatively low levels of ZAP-70. Secondly, I plan to take advantage of the increased sensitivity of Th17 cells to ZAP-70 inhibition to identify TCR signaling pathways that are most sensitive to ZAP-70 inhibition and thus highly important for Th17 differentiation. Aim 2 will determine whether incomplete or transient inhibition of ZAP-70 results in subsequent T cell hyporesponsiveness and identify potential mechanisms that drive this process. Lastly, the goal of Aim 3 is to determine the efficacy of ZAP-70 inhibition as a therapy to either prevent or ameliorate pathology associated with collagen-induced arthritis, and experimental autoimmune encephalomyelitis, two mouse models of Th17-mediated autoimmune disease. I anticipate that these studies will show for the first time that ZAP-70 inhibition effectively inhibits T cell-associated pathology in settings of autoimmunity. Ultimately, these studies may provide new insights into how T cell receptor signaling can alter the differentiation and effector functions ofT cells, which will be important for developing novel therapies for T cell-mediated autoimmune disease.

描述（由申请人提供）：候选人和培训环境：我正在寻求K 01辅导职业发展奖，以获得额外的培训，这将有助于我过渡到一个独立的科学家。我的长期研究目标是了解如何操纵T细胞受体信号可以用来改变或防止不同的T细胞功能，我的兴趣是开发T细胞介导的疾病的治疗策略。作为Deborah Fowell博士指导下的博士生，我确定了CD 4 + T细胞分化中酪氨酸激酶Itk的要求。在这些研究过程中，我获得了细胞T细胞免疫学的培训，主要涉及原代小鼠CD 4 + T细胞的分离，活化，分化和效应细胞因子的产生。我在亚瑟韦斯博士实验室的博士后培训迄今为止一直专注于开发和表征酪氨酸激酶ZAP-70的独特实验抑制剂。我们发现，这种抑制剂系统是高度特异性的，可滴定的，并且可以快速和可逆地抑制ZAP-70激酶活性，使该系统成为调节T细胞受体信号传导幅度的非常有用的工具。在我的博士后培训的这一点上，我所学到的技术和技巧非常适合于理解如何操纵T细胞受体信号传导可以应用于改变T细胞功能，这对于开发T细胞介导的自身免疫性疾病（如类风湿性关节炎）的新治疗策略可能很重要。在分析自身免疫性疾病模型方面的进一步培训，以及驱动自身反应性T细胞活化和分化的信号，将使我能够制定独立的研究计划，并转变为独立的科学家。拟议的研究将在弗朗西斯科加州大学（UCSF）的亚瑟韦斯博士的指导下进行。韦斯博士是一位风湿病学家，T细胞信号转导领域的著名专家，也是一位杰出的导师，获得了UCSF终身成就导师奖。此外，我将能够从我在UCSF的合作者/顾问Abul Abbas博士和Mark安德森博士那里获得建议，他们在自身免疫和耐受性领域拥有丰富的经验和专业知识。加州大学旧金山分校的微生物学和免疫学系也受到高度重视，是获得额外培训和自身免疫研究投入的丰富环境。研究：T细胞反应与多种自身免疫性疾病有关，如类风湿性关节炎（RA）。因此，阻断T细胞功能的治疗策略可用于治疗RA和其他T细胞介导的自身免疫性疾病。虽然抑制所有T细胞功能将阻断自身反应性T细胞应答，但更理想的策略将选择性地阻断负责引起病理的T细胞亚群。以前的工作和初步数据表明，抑制酪氨酸激酶ZAP-70可以满足这些标准。首先，常规的CD 4+和CD 8+而不是调节性T细胞的活化需要ZAP-70催化活性。其次，初步数据表明，与Th 1和Th 2分化细胞相比，经历Th 17分化的CD 4 + T细胞对ZAP-70抑制实质上更敏感。总之，这些数据表明ZAP-70抑制对于治疗由自身反应性Th 17应答介导的自身免疫性疾病（例如RA）可能非常有用。在本提案的目标1中，我的目标是确定Th 17分化细胞对ZAP-70抑制的敏感性差异。首先，我将检验Th 17细胞优先来源于表达相对低水平ZAP-70的细胞的假设。其次，我计划利用Th 17细胞对ZAP-70抑制的敏感性增加来鉴定对ZAP-70抑制最敏感的TCR信号通路，因此对Th 17分化非常重要。目的2将确定ZAP-70的不完全或短暂抑制是否会导致随后的T细胞低反应性，并确定驱动这一过程的潜在机制。最后，目的3的目标是确定ZAP-70抑制作为预防或改善与胶原诱导的关节炎和实验性自身免疫性脑脊髓炎（Th 17介导的自身免疫性疾病的两种小鼠模型）相关的病理学的疗法的功效。我预计这些研究将首次表明，ZAP-70抑制剂在自身免疫性疾病中有效抑制T细胞相关的病理。最终，这些研究可能为T细胞受体信号如何改变T细胞的分化和效应功能提供新的见解，这对于开发T细胞介导的自身免疫性疾病的新疗法将是重要的。