Cellular Immunotherapy to Combat Fibrosis and Inflammation in Systemic Sclerosis

系统性硬化症(SSC)是一种影响多器官系统的自身免疫性疾病，其特征是 慢性炎症、血管病变和纤维化。SSC的标准化死亡率没有变化 在过去的40年里有很大的变化，每三个患者中就有一个在确诊后的10年内死亡。科学的 我们提出的前提是促纤维化的巨噬细胞(M？S)是SSc的关键驱动因素。我们的假设是 靶向活化的S干细胞将为该病的治疗提供一种新的有效途径。 消除这些M？S将导致纤维化的减少。我们设计了一种新的治疗方法 用嵌合抗原受体(CAR)T细胞和定位的抗CD206CAR T细胞清除促纤维化的M？S 博莱霉素治疗减少真皮厚度和与疾病进展相关的基因表达 (BLM)小鼠皮肤纤维化模型。考虑到SSC的系统性和与内部 器官受累，我们建议确定系统给予抗CD206 CAR T细胞的疗效 用于治疗肺和皮肤纤维化。 具体目标1：确定CAR T细胞免疫疗法消除纤维蛋白原S是否会减少皮肤和肺 纤维化症。CD206-CAR-CAR T细胞免疫治疗靶向髓系亚群的耗竭及其对免疫功能的影响 皮肤和肺纤维化尚不清楚。我们将使用CITE-SEQ来鉴定皮肤和肺组织中的M？亚群 CAR T细胞治疗，并将测量胶原沉积、炎症标志物和组织厚度。 具体目标2：利用系统性纤维化模型确定抗CD206 CAR T细胞在体内的持久性。 SSC的成功治疗取决于CAR-T的坚持和效应器功能的维持。要监视 抗CD206 CAR T细胞在临床前的体内研究中，我们将建立并验证一种定量PCR方法来 检测抗CD206 CAR基因的拷贝数。

Systemic sclerosis (SSc) is an autoimmune disease that affects multiple organ systems and is characterized by chronic inflammation, vasculopathy, and fibrosis. The standardized mortality ratio for SSc has not changed significantly in the past 40 years, and one in three patients dies within 10 years of diagnosis. The scientific premise of our proposal is that pro-fibrotic macrophages (MØs) are key drivers of SSc. Our hypothesis is that targeting activated MØs in SSc will provide a novel and effective approach to treat the disease, and that the elimination of these MØs will result in decreased fibrosis. We have designed a novel therapeutic approach to eliminate pro-fibrotic MØs using chimeric antigen receptor (CAR) T cells, and localized anti-CD206 CAR T cell treatment reduces dermal thickness and gene expression associated with disease progression in a bleomycin (BLM) mouse model of skin fibrosis. Given the systemic nature of SSc and the mortality associated with internal organ involvement, we propose to determine the efficacy of systemic administration of anti-CD206 CAR T cells for the treatment of pulmonary and dermal fibrosis. Specific Aim 1: Determine if elimination of profibrotic MØs by CAR T cell immunotherapy reduces skin and lung fibrosis. Depletion of the myeloid subsets targeted by CD206-CAR T cell immunotherapy and their impact on skin and lung fibrosis are unknown. We will use CITE-seq to identify MØ subsets in skin and lung pre and post- CAR T cell treatment and will measure collagen deposition, markers of inflammation, and tissue thickness. Specific Aim 2: Determine anti-CD206 CAR T cell persistence in vivo using a systemic model of fibrosis. Successful treatment of SSc will depend on CAR T persistence and maintenance of effector function. To monitor anti-CD206 CAR T cells in preclinical in vivo studies, we will develop and validate a quantitative PCR assay to measure copy number of the anti-CD206 CAR gene.