Precise targeting of T1D specific T cells using CAR and peptide-MHC chimeric antigen ligands

项目摘要 1型糖尿病(T1D)是一种无法治愈的自身免疫性疾病，表现为胰岛的免疫破坏 β细胞。这项建议描述了一种独特的CAR T细胞方法来消除胰岛自身反应性T细胞，因此 消除自身免疫的来源，为T1D提供高度特异性的治疗。我们将利用 我们最近开发的带有嵌合抗原配体(CAL)接头的分离式CAR系统由pMHC组成 多聚体与招募分子融合在一起。CAL的添加将与自动反应TCR结合并招募 通用CAR T细胞杀死自身反应性T1D特异性T细胞。拟议中的实验将检验这一假设- ESES表示：(1)我们的模块化汽车/CAL系统特别适合于专门针对多个汽车- 反应性TCR，以及(2)我们可以识别针对T1D特定TCR的新表位以与之配对 CAR/CAL系统。重要的是，初步数据支持这些假设，并具有良好的临床特征 在这项建议中，建立了来自T1D患者的样本和严格的实验设计，基本 跨学科合作和专业知识，包括合成生物学、大规模表位发现、 T1D免疫学，人源化T1D动物模型。这项五年计划的具体目标如下 目的1建立和优化抗人TCRs的CAL体系，目的2鉴定 T1D患者/供者TCRs的自身反应性，并鉴定针对这些TCRs的多肽-MHC，以及目标3 在人源化的T1D动物模型上评价CAL系统的性能。这项工作的成果将是 建立CAR/CAL系统作为一种安全、有效和潜在治愈的T1D治疗的临床潜力。

Project Summary Type 1 diabetes (T1D) is an incurable autoimmune disease manifested through the immune destruction of islet beta cells. This proposal describes a unique CAR T cell approach to eliminate islet-autoreactive T cells, thus eliminating the source of the autoimmunity and providing a highly specific treatment for T1D. We will leverage our recently developed split CAR system with a chimeric antigen ligand (CAL) adaptor consisting of a pMHC multimer fused to a recruitment molecule. The addition of CAL will bind to the autoreactive TCR and recruit the universal CAR T cells to kill autoreactive T1D specific T cells. The proposed experiments will test the hypoth- eses that (1) our modular CAR/CAL systems are uniquely suited for specifically targeting multiple auto- reactive TCRs, and (2) that we can identify novel epitopes against T1D specific TCRs to be paired with the CAR/CAL system. Importantly, preliminary data support these hypotheses, and well-characterized clinical specimens from T1D patients and rigorous experimental designs are established in this proposal with essential cross-disciplinary collaborations and expertise that encompass synthetic biology, large-scale epitope discovery, T1D immunology, and humanized T1D animal modeling. The specific aims of this five-year proposal are as follows: Aim 1 establish and optimize the CAL system against antigen-specific human TCRs, Aim 2 characterize the autoreactivity of TCRs from T1D patients/donors and identify peptide-MHC against these TCRs, and Aim 3 evaluate the performance of the CAL system in humanized T1D animal models. Outcomes from this work will establish the clinical potential of the CAR/CAL system as a safe, effective, and potentially curative T1D treatment.