[Single-cell transcriptomic analysis reveals immune dysregula-tion and macrophage reprogramming in diabetic foot ulcers].

Objectives: To elucidate the mechanisms underlying macrophage-mediated inflammation and tissue injury in diabetic foot ulcers (DFU).

Methods: Skin tissue samples were collected from patients with diabetic foot ulcers and with non-ulcer diabetic foot (NDFU). A total of 79 272 high-quality cell transcriptomes were obtained using single-cell RNA sequencing. An unbiased clustering approach was employed to identify cell subpopulations. Systematic comparative analyses between the DFU and NDFU groups were conducted, including Gene Ontology (GO) enrichment analysis and screening of differentially expressed genes. Furthermore, cell-cell communication network construction and ligand-receptor interaction analysis were integrated to reveal the mechanisms underlying cellular interactions and signaling regulation in the DFU microenvironment from multiple perspectives.

Results: The results revealed a significant expansion of myeloid cells in DFU tissues, alongside a marked reduction in structural cells such as endothelial cells, epithelial cells, and smooth muscle cells. Major cell types underwent functional reprogramming, characterized by immune activation and impaired tissue remodeling. Specifically, macrophages in DFU tissues exhibited a shift toward a pro-inflammatory M1 phenotype, with upregulation of genes associated with inflammation and oxidative stress. Cell communication analysis further demonstrated that M1 macrophages act as both primary signal receivers and influencers in the COMPLEMENT pathway, and as key signal senders and regulators in the SPP1 pathway, actively shaping the inflammatory microenvironment. Key ligand-receptor interactions driving macrophage signaling were identified, including C3-(ITGAM+ITGB2) and SPP1-CD44.

Conclusions: This study establishes a comprehensive single-cell atlas of DFU, revealing the role of macrophage-driven cellular networks in chronic inflammation and impaired healing. These findings may offer potential novel therapeutic targets for DFU treatment.