Cellular Immunotherapy to Combat Fibrosis and Inflammation in Systemic Sclerosis

系统性硬化症（SSc）是一种影响多个器官系统的自身免疫性疾病，其特征在于： 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，有三分之一的患者在诊断后10年内死亡。科学 我们的建议的前提是促纤维化巨噬细胞（MHP）是SSc的关键驱动因素。我们的假设是 靶向SSc中活化的MMPs将提供一种新的有效的治疗疾病的方法， 消除这些MIBs将导致纤维化减少。我们设计了一种新的治疗方法， 使用嵌合抗原受体（CAR）T细胞和定位的抗CD 206 CAR T细胞消除促纤维化MSTs 治疗降低了博来霉素中与疾病进展相关的真皮厚度和基因表达 (BLM)小鼠皮肤纤维化模型。考虑到SSc的全身性质和与内分泌相关的死亡率， 器官受累，我们建议确定抗CD 206 CAR T细胞全身给药的疗效 用于治疗肺和皮肤纤维化。 具体目标1：确定通过CAR T细胞免疫疗法消除促纤维化MMPs是否会减少皮肤和肺 纤维化CD 206-CAR T细胞免疫疗法靶向的髓系亚群的耗竭及其对免疫功能的影响 皮肤和肺纤维化是未知的。我们将使用CITE-seq来鉴定治疗前和治疗后皮肤和肺中的MMP 3亚群。 CAR T细胞治疗，并将测量胶原蛋白沉积，炎症标志物和组织厚度。 具体目的2：使用全身性纤维化模型确定抗CD 206 CAR T细胞在体内的持久性。 SSc的成功治疗将取决于CAR T的持久性和效应子功能的维持。监测 在临床前体内研究中，我们将开发并验证定量PCR检测， 测量抗CD 206 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.