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

项目总结 这项建议旨在开发免疫细胞工程方法来监测和治疗 T细胞介导的自身免疫性疾病，如I型糖尿病(T1D)。天然的5-模块受体 推动T细胞对嵌入在MHC分子中的多肽抗原的反应的复合体通过 在过去约4.35亿年的迭代过程中，优化T细胞对广泛的 挑战。利用仿生学的方法，我们开发了一种5模块嵌合抗原受体(5MCAR) 并报道了表达5MCAR的细胞毒性T细胞(5MCAR-CTL)可以特异性地靶向和杀伤 自身免疫的CD4+T细胞在临床前小鼠模型中介导T1D，导致显著降低 5MCAR-CTL处理小鼠的发病率。我们的概念验证工作支持这一想法，得到了 我们正在回应的RFA，基于免疫细胞的工程化免疫疗法有望为 自身免疫性疾病的治疗。此外，正如《框架协议》所指出的，这一领域的调查尚处于起步阶段， 需要探索性工程方法的支持才能充分发挥其潜力。我们的基础性工作 为我们提供了一个继续开发新型仿生工程的新平台 检测和消除自身免疫T细胞反应的方法。我们在这里的目标是发展 使用我们的第一代5MCAR技术的新型应用程序，并通过 第二代5MCARs的工程和测试。具体来说，我们将：1)确定是否使用5MCAR重定向 中央记忆CD8+T细胞、干细胞记忆CD8+T细胞或Tregs在预防T1D方面最有效 临床前模型；2)使用5MCAR-CTL作为可以在全身贩运的哨兵，并报告 在临床症状出现之前存在致病的自身免疫的CD4+T细胞；以及，3)第二次设计和测试 5代MCAR更接近于其本地同行，目标是改进5MCAR 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5MCAR-CTL和5MCAR-Tregs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.