ELF3-regulated PES1 targets VEGFR2 to mediate angiogenesis and retinal inner barrier injury in diabetic retinopathy.

BACKGROUND: Angiogenesis and increased vascular permeability caused by endothelial cell dysfunction are crucial in diabetic retinopathy (DR). Pescadillo ribosomal biogenesis factor 1 (PES1) plays key roles in diabetes; however, its effect on DR has not been thoroughly investigated. This study evaluated the role of PES1 in angiogenesis and blood-retinal barrier functions of DR and explored the related regulatory mechanisms. METHODS: Integrated bulk RNA-seq and single-cell RNA-seq analyses were used to identify the key molecular target PES1 in this study. Western blot, scratch assay, tube formation assay, transwell, EdU proliferation assay, immunofluorescence, Evans Blue assay, PAS staining and H&E staining were used to detect the role of PES1 in angiogenesis and retinal barrier injury in vivo and in vitro, respectively. ChIP-seq was used to explore the regulatory mechanism. RESULTS: Integrated bulk RNA-seq and single-cell RNA-seq analyses revealed that high PES1 expression was significantly associated with DR. Under hyperglycemia conditions, elevated PES1 expression disrupted the retinal vascular barrier function and exacerbated vascular leakage and DR progression by inhibiting the expression of VE-cadherin and Occludin. Moreover, PES1 aggravated the imbalance between oxidation and anti-oxidation, leading to an excessive release of reactive oxygen species and further impairment of endothelial cell function. These biological processes were reversed by PES1-targeted siRNA/shRNA. Mechanistically, PES1 bound to the enhancer of VEGFR2 to regulate VEGFR2 mRNA expression, thereby influencing tube formation capacity and permeability of endothelial cells. Under diabetic conditions, PES1 upregulation was regulated by ELF3 binding to its promoter. CONCLUSIONS: Our study confirmed that the ELF3/PES1/VEGFR2 signaling pathway is crucial for regulating retinal angiogenesis and blood-retinal barrier function in DR. Interventions targeting ELF3 or PES1 may serve as potential therapeutic strategies for DR.