Multifactor mRNA Mediated T Cell Reprogramming for Systemic Lupus Erythematosus

项目摘要 系统性红斑狼疮（SLE）是一种严重的系统性自身免疫性疾病， 发病率和死亡率约为1至2百万美国人。SLE发病机制中的关键步骤是 免疫耐受性的破坏，导致T细胞依赖性和B细胞介导的疾病。的 由自身反应性B细胞产生的大量致病性自身抗体是 SLE。在B细胞缺陷的SLE易感小鼠中消除疾病，验证了它们作为一种可行的治疗方法 目标事实上，几种不同的B细胞定向疗法已经成功地减弱了进展， 包括抗CD20抗体利妥昔单抗和抗BAFF（BLyS）抗体贝利木单抗-第一个FDA 治疗系统性红斑狼疮的药物尽管这些令人兴奋的发展，有限的临床反应，只有在 一半的治疗患者，需要进一步探索B细胞耗竭的程度和功效。整体 本申请的目的是使用CAR-T细胞优化SLE的B细胞靶向治疗干预。 使用表达嵌合抗原受体（CAR）的遗传修饰的T细胞的连续性免疫疗法具有以下优点： 在杀死选定的细胞群体方面显示出显著的治疗成功。特别是，识别 CD 19对以前无反应的B细胞恶性肿瘤显示出令人印象深刻的疗效，甚至 浆细胞肿瘤在这里，我们建议使用我们的多重mRNA介导的T细胞重编程， 瞬时根除异常CD 19阳性B细胞的技术。与抗体相比，细胞 免疫治疗具有主动组织渗透和不需要FcR介导的摄取的优点， 在SLE中被阻断。与T细胞的DNA重编程相比，mRNA方法将允许T细胞的DNA重编程。 短暂的B细胞耗竭，可用于匹配SLE的复发-缓解性质。我们将测试 假设我们的mRNA方法将通过引入T细胞来改善受损SLE T细胞的功能， 基因，编码多种有益的蛋白质在一个单一的快速步骤。目标1将纠正SLE T的倾向 通过用编码抗凋亡蛋白的mRNA的组合重编程使细胞经历凋亡 除了CD19 CAR。目的2将通过一种编码T细胞代谢稳定性的mRNA来改善SLE T细胞的代谢稳定性。 除了CD19 CAR之外，SLE中减少的生长刺激细胞因子。Aim 3将重新程控SLE 具有CD19 CAR和趋化因子受体的T细胞，该趋化因子受体将CAR T细胞引导至富含B细胞的区域。我们 我们将在早期和已确诊的小鼠SLE以及 SLE患者的淋巴细胞。CAR-T方法在SLE中的成功将是自身免疫性疾病的新方法。 疾病治疗，并将导致许多潜在的机会，治疗探索。

PROJECT SUMMARY Systemic Lupus Erythematosus (SLE) is a severe systemic autoimmune disease that leads to substantial morbidity and mortality in approximately 1 to 2 million Americans. A critical step in the pathogenesis of SLE is a breakdown in immune tolerance, which leads to a T-cell dependent and B-cell mediated disease. The resulting abundant pathogenic autoantibodies produced from the autoreactive B cells are a defining hallmark of SLE. The abrogation of disease in SLE-prone mice deficient in B cells, validates them as a viable therapeutic target. Indeed, several different B-cell directed therapies have been successful at attenuating progression including the anti-CD20 antibody Rituximab, and the anti-BAFF (BLyS) antibody belimumab – the first FDA approved drug for SLE in decades. Despite these exciting developments, the limited clinical response in only half of the treated patients, calls for further exploration of the extent and efficacy of B cell depletion. Overall the goal of this application is to optimize a B-cell targeted therapeutic intervention for SLE using CAR-T cells. Adoptive immunotherapy with genetically modified T cells expressing Chimeric Antigen Receptors (CARs) has shown remarkable therapeutic success in killing select cell populations. In particular, CARs that recognize CD19 have displayed impressive efficacy toward previously unresponsive B cell malignancies, and even plasma cell neoplasms. Here, we propose to employ our multiplexed mRNA-mediated T cell reprogramming technology to transiently eradicate aberrant, CD19-positive B cells. In comparison to antibodies, cellular immunotherapy has the advantages of active tissue penetration and no requirement for FcR-mediated uptake, which is blocked in SLE. In comparison to DNA reprogramming of T cells, an mRNA approach will permit a transient B cell depletion that can be deployed to match the relapsing-remitting nature of SLE. We will test the hypothesis that our mRNA approach will improve the functionality of the damaged SLE T cells by introducing mRNAs, coding multiple beneficial proteins in a single rapid step. Aim 1 will correct the propensity of SLE T cells to undergo apoptosis by reprogramming with a combination of mRNAs encoding an anti-apoptotic protein in addition to the CD19 CAR. Aim 2 will improve SLE T cell metabolic stability with an mRNA encoding a growth stimulatory cytokine that is diminished in SLE in addition to the CD19 CAR. Aim 3 will reprogram SLE T cells with the CD19 CAR and a chemokine receptor that will direct the CAR T cells to B cell rich areas. We will test these individual factors with complementary studies in early and established murine SLE as well as in lymphocytes from patients with SLE. Success of a CAR-T approach in SLE would be novel for autoimmune disease therapy and would lead to many potential opportunities for therapeutic exploration.