Identification of M2 Macrophage-Related Key Genes in Advanced Atherosclerotic Plaques by Network Based Analysis

Abstract

Atherosclerotic plaque accounts for major adverse cardiovascular events due to its vulnerability. The M1 and M2 macrophages are implicated in progression and regression of plaque, respectively. However, the therapeutic targets related M2 macrophages still remain largely elusive. In this study, CIBERSORT and WGCNA algorithms were employed to establish a weighted gene co-expression network for identifying M2 macrophage-related hub genes using GSE43292 dataset. The results shown that genes were classified into 7 modules, with the blue module (Cor = 0.67, P = 3e-05) being the one that was most related to M2 macrophages infiltration in advanced plaques, and then 99 hub genes were identified from blue modules. Meanwhile, 1289 differentially expressed genes (DEGs) were produced in GSE43292 dataset. Subsequently, the intersection genes of hub genes and DEGs, including AKTIP, ASPN, FAM26E, RAB23, PLS3, and PLSCR4, were obtained by Venn diagrams and named as key genes. Further validation using datasets GSE100927 and GSE41571 shown that 6 key genes all down-regulated in advanced and vulnerable plaques compared with early and stable plaque samples (|Log2 (fold change) | > 0.5, P < 0.05 or 0.001), respectively. ROC curve analysis indicated the 6 key genes might have potential diagnostic value. The validation of key genes in model in vitro and in vivo also demonstrated decreased mRNA expressions of AKTIP, ASPN, FAM26E, RAB23, PLS3, and PLSCR4 (P < 0.05 or 0.001). Collectively, we identified AKTIP, ASPN, FAM26E, RAB23, PLS3, and PLSCR4 as M2 macrophages related key genes during atherosclerotic progression, proposing potential intervention targets for advanced atherosclerotic plaques.