Chimeric antigen receptor (CAR) therapy for autoimmune arthritis

嵌合抗原受体（CAR）疗法治疗自身免疫性关节炎

• 批准号: 9295586
• 负责人: Edward F Rosloniec
• 金额: $ 13.86万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9295586
• 关键词: Adoptive Transfer; Adverse effects; Alpha Cell; Antigens; Arthritis; Asses; Autoantibodies; Autoimmune Diseases; Autoimmune Process; Autoimmune Responses; Autoimmunity; B-Lymphocytes; Biological Assay; CD28 gene; CD3 Antigens; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; CD8B1 gene; Complementary DNA; Complex; DR1 gene; Development; Disease; Engineering; Event; Flow Cytometry; Generations; Genetic Engineering; Goals; HLA-DR Antigens; HLA-DR1 Antigen; HLA-DRB1; Histology; Immunization; Immunofluorescence Immunologic; Lead; Link; Longevity; Lymphoid; MHC Class II Genes; Measurement; Measures; Mediating; Mus; Onset of illness; Organ; Peptides; Phosphorylation; Predisposition; Production; Recombinants; Rheumatoid Arthritis; Severities; Signal Transduction; Specificity; Staining method; Stains; T-Cell Proliferation; T-Lymphocyte; TNFRSF10A gene; Testing; Therapeutic; Time; Transfection; Visual; ZAP-70 Gene; autoimmune arthritis; base; chimeric antigen receptor; cytokine; experimental study; genetically modified cells; humanized mouse; in vivo; innovation; killings; mouse model; novel; peripheral blood; response; targeted treatment

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.

摘要 该提案的重点是开发新型嵌合抗原受体（CAR）T 特异性靶向和消除CD 4+自身免疫T细胞的细胞， 改变自身免疫性关节炎的病程。这种创新的方法是基于 产生经遗传工程化以表达负载的HLA-DR CAR分子的CD 8 + T细胞 与自身抗原肽结合使用我们充分表征的HLA-DR人源化小鼠模型 类风湿性关节炎（RA），我们将测试的假设，CD 8 + T细胞可以工程化， 表达具有CD 28/CD 3+细胞内信号结构域的HLA-DR II类CAR，并且在 当CAR DR分子的肽负载在这些CD 8 + T细胞上时，它们将起到CTL的作用， 特异性靶向并消除CD 4+自身免疫T细胞。将采取两种办法， 产生CD 4 + T细胞的抗原特异性靶向。初步实验将集中在 使用编码与DR 1分子共价连接的抗原肽的DRB 1 cDNA。 该方法将用于证明CAR CTL生成的可行性及其在细胞中的应用。 在体内抑制活动性自身免疫性疾病的功能。在第二种方法中，我们 将产生具有“空“DR 1分子的CAR CTL，并且可溶性肽将用于负载 在测量CAR CTL功能之前检测DR 1 CAR。这种方法将允许有针对性地 CD 4 + T细胞特异性针对免疫后修饰（PTM）肽，所述肽是 小鼠模型的自身免疫性和RA的潜在重要组分 自身免疫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 CD 28/CD 3结构域接收信号的能力，以及它们的 在治疗活动性自身免疫性疾病中的功效。