Effect of CAR-M2 hydrogel treatment on the expression of related genes such as fibrosis and endothelial angiogenesis in mice with renal fibrosis (bulk RNA-seq)

Currently there is no effective treatment for renal fibrosis, which is the common outcome of chronic kidney disease (CKD). The renal fibrosis was aggravated by the unfavorable microenvironment and cell-to-cell interactions, accompanied by fibroblast activation and vascular rarefaction. Here, M2 macrophage engineered with chimeric antigen receptor (CAR) was created to promote renal revascularization for the synergistic treatment of phagocytosis of activated fibroblast. To achieve efficient delivery of parenchymal organs, a hydrogel was used to deliver CAR-M2 by renal subcapsule injection. CAR-M2 significantly reduced renal fibrosis and promoted revascularization. The single-cell RNA sequencing (scRNA-seq) revealed the heterogenicity and interaction of stroma and endothelial cells (ECs). A fibrosis-related Cxcr2+ EC subset was defined, which induced renal fibrosis by upregulating lipocalin 2 (Lcn2). Combined analysis of CKD patients demonstrated that CAR-M2 could release Matrix Metalloproteinase 2 (MMP2) in close proximity to activate the Cxcr2+ EC retinoid X receptor alpha (Rxra) to regulate fibrosis. Our study provides an effective strategy to develop an advanced cell therapy for organ fibrosis.