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

项目摘要 1型糖尿病（T1 D）是一种无法治愈的自身免疫性疾病，表现为胰岛细胞的免疫破坏， β细胞该提案描述了一种独特的CAR T细胞方法来消除胰岛自身反应性T细胞， 消除了自身免疫的来源，并为T1 D提供了高度特异性的治疗。我们将利用 我们最近开发的具有嵌合抗原配体（CAL）衔接子的分裂CAR系统由pMHC 多聚体与募集分子融合。CAL的添加将结合自身反应性TCR并募集免疫应答。 通用CAR T细胞杀死自身反应性T1 D特异性T细胞。这些实验将检验假设- （1）我们的模块化CAR/CAL系统是唯一适合专门针对多个汽车， 反应性TCR，以及（2）我们可以鉴定针对T1 D特异性TCR的新表位，以与 CAR/CAL系统。重要的是，初步的数据支持这些假设， T1 D患者的标本和严格的实验设计建立在这个建议与基本的 跨学科合作和专业知识，包括合成生物学，大规模表位发现， T1 D免疫学和人源化T1 D动物建模。这一五年计划的具体目标是： 目的1建立并优化针对抗原特异性人TCR的CAL体系，目的2表征 来自T1 D患者/供体的TCR的自身反应性和鉴定针对这些TCR的肽-MHC，以及Aim 3 评价CAL系统在人源化T1 D动物模型中的性能。这项工作的成果将 确立CAR/CAL系统作为安全、有效和潜在治愈性T1 D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.