Integrated multi-omic high-throughput strategies across-species identified potential key diagnostic, prognostic, and therapeutic targets for atherosclerosis under high glucose conditions

Diabetes is a well-known risk factor for atherosclerosis (AS), but the underlying molecular mechanism remains unknown.Hyperglycemia is a hallmark of diabetes and high glucose is proven to induce foam cell formation mainly derived from macrophages. In this study, we aimed to explore systematic regulatory gene programs under this complicated situation. Following by induction of macrophage-derived foam cells in vitro, the high-throughput sequencing was performed. Coexpression gene modules were constructed using weighted gene co-expression network analysis (WGCNA) and relevant modules were identified. Genes with high intramodular connectivity were further screened to select hub genes related to foam cell formation in foam cells annotated by integrated scRNA datasets. The potential roles of selected genes were further validated in bulk-RNA and scRNA datasets of human plaques. Two modules were found to be both positively related to high glucose and ox-LDL. Further enrichment analyses confirmed the association between the brown module and AS. The high correlation between the brown module and macrophages was identified and 4 genes (Aldoa, Creg1, Lgmn, and Pkm) were screened. Further validation in external bulk-RNA and scRNA datasets of human plaques was performed. In addition, the survival analysis confirmed the prognostic value of Aldoa. Knocking down Aldoa expression alleviated the foam cell formation in vitro. Overall design: Mice aged 8 weeks were intraperitoneally injected with 6% soluble starch and the peritoneal macrophages (PM) were isolated. For foam cell induction, the isolated peritoneal macrophages were incubated with 50?mg/ml ox-LDL (Yiyuan Biotech, China) for 48?h. As for high glucose induction, the cells were treated with high glucose (33mmol/L) medium, while mannitol (27.5mM) was added in DMEM as an osmotic control. 4 groups were set, low glucose without ox-LDL (L_C), high glucose without ox-LDL (H_C), low glucose with ox-LDL (L_O), high glucose with ox-LDL (H_O) The mouse strain (background) used in these experiments is C57BL/6.