Titrating CD45 expression in mice to study development of T cell tolerance

滴定小鼠 CD45 表达以研究 T 细胞耐受的发展

• 批准号: 7952505
• 负责人: JULIE ZIKHERMAN
• 金额: $ 11.96万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7952505
• 关键词: Address; Advisory Committees; Affect; Animals; Antigen Receptors; Arts; Autoantigens; Autoimmune Diseases; Autoimmune Polyendocrinopathies; Autoimmunity; Biological; Bone Marrow; Celiac Disease; Cells; Characteristics; Chimera organism; Chronic Disease; Clinical; Data; Development; Developmental Biology; Disease; Dose; Environment; Equilibrium; Exhibits; Foundations; Future; Genes; Genetic; Genetic Polymorphism; Goals; Grant; Host Defense; Human; Human Genetics; Immune Tolerance; Immune system; Immunologic Deficiency Syndromes; Immunology; Individual; Insulin-Dependent Diabetes Mellitus; Investigation; Investments; Journals; Laboratories; Light; Link; Medicine; Mentors; Molecular; Molecular Immunology; Morbidity - disease rate; Mus; Myasthenia Gravis; Organ; Organ Model; Output; PTPRC gene; Pathogenesis; Peripheral; Phenotype; Phosphoric Monoester Hydrolases; Phosphotransferases; Physiological; Play; Positioning Attribute; Process; Radio; Receptor Signaling; Regulation; Research; Research Personnel; Research Proposals; Resistance; Rheumatoid Arthritis; Rheumatology; Role; Scientist; Series; Signal Transduction; Sorting - Cell Movement; Specificity; Staining method; Stains; Surface; Syndrome; System; T-Cell Development; T-Cell Receptor; T-Lymphocyte; Thymic epithelial cell; Thymocyte Selection; Thymus Gland; Titrations; Training; Transcriptional Regulation; Work; analog; autoreactive T cell; career; career development; central tolerance; design; digital; experience; genetic risk factor; meetings; molecular phenotype; mortality; novel; novel strategies; novel therapeutics; professor; programs; research study; thymocyte; treatment strategy

DESCRIPTION (provided by applicant): This proposal describes a five year career development program whose goal is to prepare Dr. Julie Zikherman for a career as an independent investigator. This program will build on Dr. Zikherman's experience in clinical rheumatology, basic developmental biology and basic immunology in order to expand an emerging research program and provide her with further expertise in the design and conduct of cellular and molecular immunology research. The long-term goal is to establish an independent laboratory dedicated to the study of basic mechanisms of autoimmune disease pathogenesis, with a focus on how antigen receptor signaling can establish and subvert immune tolerance. The central guidance and training environment for this project will be provided by the mentor, Dr. Art Weiss, Engleman Distinguished Professor of Medicine, and Division Chief of Rheumatology at UCSF, as well as an expert in T cell receptor signaling. The training plan includes formal course work and didactic training through attendance of seminar series, journal clubs, UCSF retreats and scientific meetings, as well as guidance by an advisory committee of established scientists, and a research program that will provide additional technical training along with new directions for independent investigation. The studies proposed in this grant are intended to elucidate how altered T cell receptor signaling can subvert mechanisms of central tolerance and produce autoimmunity in humans. In her preliminary data, Dr. Zikherman has generated an allelic series of mice expressing distinct levels of CD45, and demonstrated that varying CD45 dose produces stable titration of TCR signaling intensity as well as quantitative changes in thymic output. In the research proposal, Dr. Zikherman will take advantage of this novel system to study how titration of TCR signaling intensity regulates thymic cell fate, transcriptional machinery and ultimately functional T cell repertoire. In the first part of the proposal, Dr. Zikherman will use genetic approaches and bone marrow chimeras to define the regulation of positive and negative selection in the allelic series. In the second part she will characterize the molecular phenotype of allelic series thymocytes by defining expression of key transcriptional regulators of positive selection. In the final part of the proposal, Dr. Zikherman will capitalize on the ability to track individual autoreactive cells in the context of the Aire-/- model of organ-specific autoimmunity to study how TCR signaling affects functional T cell repertoire. Together, these experiments will serve to characterize the cellular, molecular, and functional consequences of altered TCR signaling during T cell development. These studies will illuminate the mechanisms by which altered TCR signaling undermines central tolerance to produce disease. In addition to addressing the questions posed in this research proposal, the studies outlined above will serve as a foundation for future work that will enable Dr. Zikherman to become an independent investigator.

描述(由申请人提供)：本提案描述了一项为期五年的职业发展计划，其目标是为朱莉·齐克曼博士作为一名独立研究员的职业生涯做好准备。该项目将以Zikherman博士在临床风湿学、基础发育生物学和基础免疫学方面的经验为基础，以扩大新兴的研究项目，并为她提供细胞和分子免疫学研究设计和实施方面的进一步专业知识。长期目标是建立一个独立的实验室，致力于研究自身免疫性疾病发病的基本机制，重点是抗原受体信号如何建立和颠覆免疫耐受。该项目的中央指导和培训环境将由导师Art Weiss博士提供，他是加州大学旧金山分校的英国杰出医学教授和风湿病科主任，以及T细胞受体信号方面的专家。培训计划包括通过参加研讨会系列、期刊俱乐部、加州大学旧金山分校的务虚会和科学会议，以及由知名科学家组成的咨询委员会的指导，以及将提供额外技术培训和独立调查新方向的研究计划，进行正式的课程工作和教学培训。这项资助中提出的研究旨在阐明改变的T细胞受体信号如何能够颠覆中枢耐受机制，并在人类中产生自身免疫。在她的初步数据中，齐克曼博士产生了一系列表达不同水平CD45的等位基因小鼠，并证明了不同的CD45剂量会产生稳定的TCR信号强度滴定以及胸腺输出的数量变化。在这项研究提案中，齐克曼博士将利用这一新系统来研究TCR信号强度的滴定如何调节胸腺细胞的命运、转录机制以及最终的功能T细胞库。在该提案的第一部分，齐克曼博士将使用遗传学方法和骨髓嵌合体来定义等位基因序列中正选择和负选择的调控。在第二部分中，她将通过定义正选择的关键转录调节因子的表达来表征等位基因系列胸腺细胞的分子表型。在该提案的最后部分，齐克曼博士将在器官特异性自身免疫的Aire-/-模型的背景下，利用追踪单个自身反应细胞的能力，研究TCR信号如何影响功能T细胞谱。总之，这些实验将用于描述T细胞发育过程中TCR信号改变的细胞、分子和功能后果。这些研究将阐明TCR信号改变破坏中枢耐受性导致疾病发生的机制。除了解决本研究提案中提出的问题外，上述研究还将作为未来工作的基础，使齐克曼博士能够成为一名独立的调查员。