ANGPTL4-dependent metabolic reprogramming fuels RhoA signalling and microvascular dysfunction in diabetes.

backgroundDiabetic microvascular complications (DMCs) are a principal driver of blindness, kidney failure, neuropathy, and non-healing ulcers. A shared early lesion across microvascular beds is endothelial barrier breakdown with pathological hyperpermeability, which is exacerbated by diabetic metabolic stress and heterogeneous microcirculatory flow. The molecular nodes that connect flow-derived mechanical cues to metabolic remodeling and sustained permeability remain incompletely defined.

methodsWe implemented an integrated multi-omics strategy combining bulk transcriptomics, network biology, and spatially resolved single-cell analyses. Bulk RNA-sequencing datasets from ApoE−/− murine aortas and human carotid/aortic lesion tissues were analyzed using differential expression, weighted gene co-expression network analysis (WGCNA), and pathway enrichment. Single-cell RNA-sequencing (GSE159677) was used to resolve cell-type-specific expression patterns and ANGPTL4-stratified endothelial states. Spatial transcriptomics (GSE241346; Visium) localized ANGPTL4 and RhoA within lesion microenvironments. Finally, protein-compound interaction profiling and AutoDock Vina molecular docking were used to prioritize candidate modulators, focusing on Tanshinol B.

resultsAngiopoietin-like 4 (ANGPTL4) was consistently downregulated and occupied central positions in co-expression modules enriched for endothelial barrier regulation, cytoskeletal remodeling, and mitochondrial bioenergetic pathways. Across independent cohorts, reduced ANGPTL4 expression was robustly associated with increased RhoA expression and with transcriptional programs consistent with heightened actomyosin contractility and endothelial permeability, accompanied by signatures of mitochondrial dysfunction and metabolic reprogramming. Single-cell and spatial analyses concentrated ANGPTL4 expression within endothelial compartments and supported a spatial association between ANGPTL4-low regions and RhoA-high transcriptional features. In silico docking predicted that Tanshinol B engages specific residues on both ANGPTL4 and RhoA, suggesting dual-target potential.

conclusionsThese results define a conserved ANGPTL4-RhoA axis coupled to metabolic dysregulation and barrier-relevant cytoskeletal programs. We propose a mechanometabolic model in which reduced ANGPTL4 permissively enhances RhoA-driven contractility and endothelial hyperpermeability-mechanisms that are highly relevant to DMC pathogenesis. Together, our findings highlight ANGPTL4 and its pharmacologic modulation as candidate leverage points to preserve endothelial integrity in diabetes-associated microvascular disease.