Decoding the Spatiotemporal Dynamics of ENTPD1 in Diabetic Foot Ulcer Through Multi‐Omic Profiling

ABSTRACT The global incidence of diabetic foot ulcers (DFUs) has increased in parallel with the global incidence of diabetes mellitus, leading to a range of pathophysiologic conditions and a poor prognosis. ENTPD1, also known as CD39, is an extracellular nucleotidase associated with immunogenic cell death (ICD), but its role in DFUs remains unclear. Through single‐cell RNA sequencing, we obtain the genetic, functional, trajectory, and communication differences between DFUs and healthy subjects. Then, we selected vascular endothelial cells (Vasendo) for further re‐clustering to identify DFU‐healing‐promoted ENTPD1+ Vasendo and related ligand‐receptor pairs in cell communication. The study demonstrated that ICD level and ENTPD1 expression were strongly correlated with DFUs, exhibiting greater enrichment in the Healing DFUs group than the Non‐healing group. The Vasendo in the Healing group were enriched in the healing of DFUs and exhibited complex cellular communication. Re‐clustering of Vasendo further identified the ENTPD1+ subtype. The trajectory and cell communication analysis further confirmed the healing‐promoting effects of ENTPD1+ Vasendo, with inhibited ACKR1 receptor and MIF ligand when communicating with macrophages. Finally, expression quantitative trait loci (eQTL) Mendelian randomization and bulk sequencing validation data confirmed that upregulated expression of ANKIB1 and ANP32E with downregulated expression of SLC30A7 and TMF1 genes in DFUs Vasendo increased the risk of secondary diabetic peripheral artery disease (DPAD). Our study demonstrated the molecular mechanisms underlying DFUs healing and secondary DPAD occurrence, providing potential therapeutic targets for DFUs and DPAD therapy.