Stem cell-derived regulatory T cells for therapeutic use in arthritis

项目总结 CD4调节性T细胞(Tregs)对于正常的免疫监测是必不可少的，它们的功能障碍可能导致 自身免疫性疾病的发展，如类风湿性关节炎。多能干细胞(PSCs)可以 用来获得健康树突状细胞的可再生来源来治疗自身免疫性关节炎，因为它们有能力 产生体内几乎所有类型的细胞，包括树突状细胞。然而，发展的合适条件 PSC的抗原(Ag)特异性Tregs(PSC-Tregs)还没有完全定义，尤其是信号转导 指导这种树的分化的机制。抗原特异性PSC-Tregs可以是组织相关的和 渗透到局部炎症组织(例如滑膜)以抑制过继转移后的自身免疫反应， 从而避免了来自非特异性Treg的潜在的整体免疫抑制。我们的长期目标是 研究计划是开发和优化策略，以利用PSCs作为高活性Tregs的来源 基于细胞的疗法。本申请的目的是确定银离子的潜在机制。 旨在调节自身免疫性关节炎耐受性的特定PSC-Treg治疗。中心假设是 AUTO抗原特异性的PSC-Tregs是组织相关的Tregs，积聚在发炎的滑膜和 抑制过继移植后的自身免疫，是细胞治疗的极佳候选者。 在发表的结果和申请人实验室的初步数据的指导下，这一假设将得到验证 通过追求三个具体目标：1)定义汽车银特异性PSC-Tregs的基本属性，该属性 与抑制活动有关；2)确定指导自体分化的信号机制 利用自身抗原特异性PSC-Tregs研究自身免疫性关节炎的细胞治疗。 特雷格斯。在第一个目标下，体外和体内方法已被确定为可行的 申请人的实验室，将被使用。AUTO-Ag特异性PSC-Tregs的基本性质 表型、特异性和功能将被定义。在第二个目标下，基因表达谱和 将对细胞内信号通路进行研究，以确定Notch-Hes1、Notch-RUNX1和 自身抗原特异性PSC-Tregs发育过程中的Noch-Survivin信号转导。在第三个目标下，那么- 特色化的动物模型将被用来优化基于细胞的自身免疫性关节炎的治疗 抗原特异的小鼠或人PSC-Tregs。这种方法是创新的，因为汽车银特定的概念 PSC-Tregs用于自身免疫性关节炎的基于细胞的治疗之前还没有被探索过。建议数 这项研究意义重大，因为它将为开发与组织相关的PSC-Tregs奠定基础 将通过累积发炎的滑膜和驱动减少过继转移后的整体免疫抑制 将治疗性PSC-Tregs用于自身免疫性关节炎的基于细胞的治疗。

PROJECT SUMMARY CD4+ regulatory T cells (Tregs) are essential for normal immune surveillance, and their dysfunction can lead the development of autoimmune diseases, such as rheumatoid arthritis. Pluripotent stem cells (PSCs) can be utilized to obtain a renewable source of healthy Tregs to treat autoimmune arthritis as they have the ability to produce almost all cell types in the body, including Tregs. However, the right conditions for the development of antigen (Ag)-specific Tregs from PSCs (i.e., PSC-Tregs) have not been fully defined, especially the signaling mechanisms that direct differentiation of such Tregs. Ag-specific PSC-Tregs can be tissue-associated and infiltrate to local inflamed tissue (e.g., synovium) to suppress autoimmune responses after adoptive transfer, thereby avoiding potential overall immunosuppression from non-specific Tregs. The long-term goal of our research program is to develop and optimize strategies for utilizing PSCs as a source of highly reactive Tregs for cell-based therapies. The objective of this application is to determine the mechanisms underlying the Ag- specific PSC-Treg treatments that aim to modulate tolerance in autoimmune arthritis. The central hypothesis is that auto Ag-specific PSC-Tregs are tissue-associated Tregs, which accumulate in inflamed synovium and suppress the autoimmunity after adoptive transfer and act as excellent candidates for cell-based therapies. Guided by published results and preliminary data from the applicant's laboratory, this hypothesis will be tested by pursuing three specific aims: 1) to define the fundamental properties of auto Ag-specific PSC-Tregs that relate to suppressive activity; 2) to determine the signaling mechanisms that direct the differentiation of auto Ag-specific PSC-Tregs; and 3) to study cell-based therapies of autoimmune arthritis using auto Ag-specific PSC- Tregs. Under the first aim, in vitro and in vivo approaches, which have been established as feasible in the applicant's laboratory, will be used. The fundamental properties of auto Ag-specific PSC-Tregs with regard to phenotype, specificity and function will be defined. Under the second aim, gene expression profile and intracellular signaling pathways will be studied to determine the critical roles of Notch-Hes1, Notch-Runx1 and Notch-survivin signaling during the development of auto Ag-specific PSC-Tregs. Under the third aim, well- characterized animal models will be used to optimize cell-based therapies of autoimmune arthritis using auto Ag-specific mouse or human PSC-Tregs. The approach is innovative, because the concept of auto Ag-specific PSC-Tregs for cell-based therapies in autoimmune arthritis has not been previously explored. The proposed research is significant, because it will provide the foundation for developing tissue-associated PSC-Tregs that will reduce overall immunosuppression after adoptive transfer by accumulating inflamed synovium and drive forward the use of therapeutic PSC-Tregs for cell-based therapies in autoimmune arthritis.