Chimeric antigen receptor (CAR) therapy for autoimmune arthritis

摘要 这项建议的重点是开发新型嵌合抗原受体(CAR)T细胞 特异性靶向和消除CD4自身免疫T细胞的细胞，并将在治疗上 改变自身免疫性关节炎的病程。这种创新的方法是基于 基因工程表达人类白细胞抗原-DR CAR分子的CD8T细胞的制备 用一种自身抗原肽。使用我们特征良好的人类白细胞抗原-DR人源化小鼠模型 对于类风湿性关节炎(RA)，我们将检验CD8 T细胞可以被工程改造成 含CD2 8/CD3ζ细胞内信号转导结构域的人白细胞抗原-DRⅡ类CAR的表达 将CAR-DR分子负载到这些CD8 T细胞上，它们将发挥CTL和 特别针对和消除CD4自身免疫T细胞。将采取两种方法来 产生针对CD4T细胞的抗原特异性靶向。最初的实验将集中在 使用编码与DR1分子共价连接的抗原肽的DRB1cDNA。 这种方法将被用来证明汽车CTL生成的可行性和他们的 在体内抑制一种活跃的自身免疫性疾病的功能。在第二种方法中，我们 将产生带有空的DR1分子的CAR CTL，并将使用可溶性多肽来加载 对DR1车前车进行CTL功能测量。这种方法将允许有针对性地 翻译后修饰(PTM)多肽的CD4T细胞，这些多肽是 类风湿性关节炎小鼠模型的自身免疫及可能的重要成分 自身免疫力。通过这项提案的具体目标，我们将调查 CAR T细胞，它们通过CAR CD28/CD3区域接收信号的能力，以及它们的 治疗一种活动性自身免疫性疾病的疗效。

Abstract The focus of this proposal is the development of novel chimeric antigen receptor (CAR) T cells that specifically target and eliminate CD4+ autoimmune T cells, and will therapeutically alter the disease course of autoimmune arthritis. This innovative approach is based on the generation of CD8+ T cells genetically engineered to express HLA-DR CAR molecules loaded with an autoantigenic peptide. Using our well characterized HLA-DR humanized mouse model of rheumatoid arthritis (RA), we will test the hypothesis that CD8+ T cells can be engineered to express HLA-DR class II CAR with CD28/CD3ζ intracellular signaling domains, and that after peptide loading of the CAR DR molecules on these CD8+ T cells, they will function as CTL and specifically target and eliminate CD4+ autoimmune T cells. Two approaches will be pursued to generate antigen specific targeting of the CD4+ T cells. Initial experiments will be focused on using DRB1 cDNA encoding an antigenic peptide that is covalently linked to the DR1 molecule. This approach will be used to demonstrate the feasibility of the CAR CTL generation and their function in vivo in the inhibition of an active autoimmune disease. In the second approach, we will generate CAR CTL with “empty “ DR1 molecules, and soluble peptide will be used to load the DR1 CAR before measuring the CAR CTL function. This approach will allow for targeting CD4+ T cells specific for posttranslationally modified (PTM) peptides which are components of the autoimmunity of the mouse model and potentially important components of RA autoimmunity. Through the specific aims of this proposal, we will investigate the specificity of the CAR T cells, their ability to receive signals through the CAR CD28/CD3 domains, and their efficacy in treating an active autoimmune disease.