Single-Cell RNA-Seq Reveals Collagen VI Antibody-Induced M2-Like Macrophages Expressing Lyve1 Reduce Atherosclerotic Plaque Area in Apoe-/- Mice

The macrophage subpopulations that have therapeutic benefits against atherosclerosis have not yet been fully understood. A single-cell RNA sequencing analysis were performed on the F4/80+ macrophages of apolipoprotein E-deficient (Apoe−/−) mice on a normal diet (ND), a high-fat diet (HFD), and a high-fat diet (HFD) with anti-CVI-mAb treatment. We analyze the diversity of M2 macrophages in atherosclerosis using single cell RNA sequencing (scRNA-seq). Four-week-old male Apoe−/− mice with C57BL/6 background were fed a high-fat diet (HFD; 21 % fat, 0.15 % cholesterol) for 20 weeks. The diet was changed to a normal diet one week before the first immunization. At week 21, the mice in the anti-CVI-mAb and HFD groups received intraperitoneal injections of anti-CVI-mAb (3 mg per 200 µL) and PBS (200 µL). C57BL/6 and Apoe-/- fed normal diet (intraperitoneal injections equal volume PBS) were control. Peritoneal cells were isolated from each group. The cell suspensions obtained individually from the peritoneal cavity were passed through a 100 µm cell strainer and were stained with anti-mouse F4/80-FITC. The total viable F4/80+ cells were selected using Beckman Coulter’s MoFlo Astrios EQ for scRNA-sequencing.