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

项目摘要 CD 4+调节性T细胞（TCD 4 + regulatory T cells，TCD 4 + T细胞）是正常免疫监视所必需的，其功能障碍可导致免疫功能低下。 自身免疫性疾病的发展，如类风湿性关节炎。多能干细胞（PSC）可以是 用于获得可再生来源的健康睾酮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TCFs。然而，发展的适当条件 来自PSC的抗原（Ag）-特异性T细胞（即，PSC-TSC 2）尚未完全定义，尤其是信令 这些机制直接分化这种T细胞。Ag-特异性PSC-T细胞可以是组织相关的， 浸润到局部发炎组织（例如，滑膜）以抑制过继转移后的自身免疫应答， 从而避免了来自非特异性T细胞的潜在的整体免疫抑制。我们的长期目标是 一项研究计划是开发和优化利用PSC作为高活性TTRs来源的策略， 基于细胞的疗法本申请的目的是确定Ag- 特异性PSC-Treg治疗旨在调节自身免疫性关节炎的耐受性。核心假设是 自体Ag特异性PSC-T细胞是组织相关的T细胞，其在发炎的滑膜中积累， 抑制过继转移后的自身免疫，并作为基于细胞的治疗的优秀候选者。 根据已发表的结果和申请人实验室的初步数据，将对这一假设进行检验 通过追求三个具体的目标：1）定义汽车Ag特异性PSC-THBE的基本性质， 2）确定指导自体免疫细胞分化的信号传导机制， Ag-特异性PSC-TCLs;和3）使用自身Ag-特异性PSC-TCLs研究自身免疫性关节炎的基于细胞的疗法。 你好在第一个目标下，体外和体内方法，已经被确定为在本领域中可行， 申请人的实验室，将被使用。研究了自载银特异性PSC-T细胞的基本性质， 将定义表型、特异性和功能。在第二个目标下，基因表达谱和 将研究细胞内信号传导途径，以确定Notch-Hes 1、Notch-Runx 1和 Notch-Survivin信号转导在自身Ag特异性PSC-T细胞发育过程中的作用在第三个目标下，嗯- 特征化的动物模型将用于优化基于细胞的自身免疫性关节炎的治疗， Ag-特异性小鼠或人PSC-T细胞。这种方法是创新的，因为汽车Ag的概念是特定的 PSC-Tclase用于自身免疫性关节炎的基于细胞的疗法之前尚未探索。拟议 这项研究意义重大，因为它将为开发组织相关的PSC-THBE提供基础， 通过积累发炎的滑膜和驱动力，将减少过继转移后的总体免疫抑制 提出了治疗性PSC-T细胞在自身免疫性关节炎中用于基于细胞的疗法的用途。

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.