DYSF promotes monocyte activation in atherosclerotic cardiovascular disease as a DNA methylation-driven gene

Abstract

Dysferlin (DYSF) has drawn much attention due to its involvement in dysferlinopathy and was reported to affect monocyte functions in recent studies. However, the role of DYSF in the pathogenesis of atherosclerotic cardiovascular diseases (ASCVD) and the regulation mechanism of DYSF expression have not been fully studied. In this study, Gene Expression Omnibus (GEO) database and epigenome-wide association study (EWAS) literatures were searched to find the DNA methylation-driven genes (including DYSF) of ASCVD. The hub genes related to DYSF were also identified through weighted correlation network analysis (WGCNA). Regulation of DYSF expression through its promoter methylation status was verified using peripheral blood leucocytes (PBLs) from ASCVD patients and normal controls, and experiments on THP1 cells and Apoe^-/- mice. Similarly, the expressions of DYSF related hub genes, mainly contained SELL, STAT3 and TMX1, were also validated. DYSF functions were then evaluated by phagocytosis, transwell and adhesion assays in DYSF knock-down and overexpressed THP1 cells. The results showed that DYSF promoter hypermethylation up-regulated its expression in clinical samples, THP1 cells and Apoe^-/- mice, confirming DYSF as a DNA methylation-driven gene. The combination of DYSF expression and methylation status in PBLs had a considerable prediction value for ASCVD. Besides, DYSF could enhance the phagocytosis, migration and adhesion ability of THP1 cells. Among DYSF related hub genes, SELL was proven to be the downstream target of DYSF by wet experiments. In conclusion, DYSF promoter hypermethylation upregulated its expression and promoted monocytes activation, which further participated in the pathogenesis of ASCVD.

Keywords: 1stExon; ABR; ABR activator of RhoGEF and GTPase; ASCVD; Cq; DEG; DM; DMG; DMP; DNA methyltransferase 1; DNMT1; DYSF; EWAS; FBG; FH; GAPDH; GSEA; HD; HDL-C; HUVEC; LDL-C; MDRE; ND; PBL; PLT; PPI; RPS27; SELL; STAT3; TC; TG; TMX1; TOM; TSS; Tm; UTR; WBC; WGCNA; WT; annealing temperature; atherosclerotic cardiovascular diseases; diabetes mellitus; differentially expressed gene; differentially methylated gene; differentially methylated probe; dysferlin; epigenome-wide association study; familial hypercholesterolemia; fasting blood glucose; gene set enrichment analysis; glyceraldehyde-3-phosphate dehydrogenase; high-density lipoprotein cholesterol; high-fat diet; human umbilical vein endothelial cell; low-density lipoprotein cholesterol; methylation dependent restriction enzyme; methylation-specific qPCR; normal diet; ox-LDL; oxidized low-density lipoprotein; peripheral blood leukocyte; platelet; protein-protein interaction; qMSP; quantification cycle; ribosomal protein S27; selectin L; signal transducer and activator of transcription 3; the first exon; thioredoxin related transmembrane protein 1; topological overlap matrix; total cholesterol; transcriptional start site; triglyceride; untranslated region; weighted correlation network analysis; white blood cell; wild type.