Engineering 2nd generation 5MCARs to monitor and treat Type-I Diabetes

项目摘要 该提案旨在开发免疫细胞工程方法，用于监测和治疗 T细胞介导的自身免疫性疾病，如I型糖尿病（T1 D）。天然5-模块受体 驱动T细胞对嵌在MHC分子中的肽抗原应答的复合物已经通过以下方式进化： 在过去的4.35亿年里，这是一个迭代的过程，以优化T细胞对广泛的免疫反应。 挑战利用仿生方法，我们开发了一种5-模块嵌合抗原受体（5 MCAR）。 并报道了表达5 MCAR的细胞毒性T细胞（5 MCAR-CTL）可以特异性靶向并杀死 在临床前小鼠模型中介导T1 D的自身免疫性CD 4 + T细胞， 在5 MCAR-CTL处理的小鼠中的疾病发生率。我们的概念验证工作支持这一想法， 我们正在回应的RFA，基于工程免疫细胞的免疫疗法有望实现 自身免疫性疾病的治疗。此外，正如RFA中所指出的，这一调查领域还处于起步阶段， 需要探索性工程方法的支持，以充分发挥其潜力。我们的基础工作 为我们提供了一个新的平台，继续发展新的仿生工程 检测和消除自身免疫性T细胞反应的方法。我们的目标是发展 使用我们的第一代5 MCAR技术的新应用，并通过 第二代5 MCR的工程和测试。具体来说，我们将：1）确定是否使用5 MCAR重定向 中央记忆性CD 8 + T细胞、干细胞记忆性CD 8 + T细胞或T细胞免疫应答在预防T1 D方面最有效。 临床前模型; 2）使用5 MCAR-CTL作为哨兵，可以在整个身体中运输并报告 在出现临床症状之前存在致病性自身免疫性CD 4 + T细胞;以及，3）工程化和测试第二阶段 第五代5 MCAR，更接近地模仿其天然对应物，目的是改善5 MCAR 性能完成后，拟议的工作将为治疗提供有价值的临床前数据。 基于进化蓝图设计的仿生5 MCAR-CTL和5 MCAR-Tcls的潜力。

PROJECT SUMMARY This proposal is aimed at developing immune cell engineering approaches for the monitoring and treatment of T cell-mediated autoimmune diseases such as Type-I diabetes (T1D). The native 5-module receptor complexes that drive T cell responses to peptide antigens embedded in MHC molecules have evolved through an iterative process over the last ~435 million years to optimize T cell responses to a broad range of challenges. Using a biomimetic approach, we have developed a 5-module chimeric antigen receptor (5MCAR) and reported that cytotoxic T cells expressing 5MCARs (5MCAR-CTLs) can specifically target and kill autoimmune CD4+ T cells that mediate T1D in preclinical mouse models, resulting in significantly reduced disease incidence in 5MCAR-CTL-treated mice. Our proof-of-concept work supports the idea, echoed by the RFA to which we are responding, that engineered immune cell-based immunotherapies hold promise for the treatment of autoimmune diseases. Also as noted in the RFA, this area of investigation is in its infancy and requires support of exploratory engineering approaches to reach its full potential. Our foundational work provides us with a novel platform from which to continue the development of novel biomimetic engineering approaches for detecting and eliminating autoimmune T cell responses. Our goals here will be to develop novel applications using our 1st generation 5MCAR technology and to iterate on the 5MCAR platform through the engineering and testing of 2nd generation 5MCARs. Specifically, we will: 1) determine if using 5MCARs to redirect central memory CD8+ T cells, stem cell memory CD8+ T cells, or Tregs are most effective at preventing T1D in preclinical models; 2) use 5MCAR-CTLs as sentinels that can trafficking throughout the body and report on the presence of pathogenic autoimmune CD4+ T cells prior to clinical symptoms; and, 3) engineer and test 2nd generation 5MCARs that more closely mimic their native counterparts with the goal of improving 5MCAR performance. When completed, the proposed work will provide valuable pre-clinical data for the therapeutic potential of biomimetic 5MCAR-CTLs and 5MCAR-Tregs that are engineered based on evolution’s blueprint.