Long Noncoding RNA PPT2-EGFL8 Regulates Pathological Retinal Neovascularization in PDR by Functioning as a Competing Endogenous RNA

Diabetic retinopathy (DR) is a common complication in patients with diabetes, and proliferative DR (PDR) has become an important cause of blindness; however, the mechanisms involved have not been fully elucidated. miRNAs and long noncoding RNAs can play an important role in DR, and they can accurately regulate the expression of target genes through a new regulatory model: competing endogenous RNAs. We isolated total RNA of extracellular vesicles (EVs) in the serum of healthy individuals and individuals with diabetes without DR, non-PDR, or PDR, and performed deep sequencing. We found aberrantly low expression of PPT2-EGFL8 and significantly increased level of miR-423-5p. PPT2-EGFL8 adsorbs miR-423-5p as a molecular sponge and inhibits hypoxia-induced human retinal microvascular endothelial cells proliferation. In an oxygen-induced retinopathy (OIR) model and a streptozotocin-induced diabetes model, Egfl8-overexpression treatment reduces diabetes-related reactive gliosis, inflammation, and acellular capillaries and attenuates the development of pathological neovascularization. In addition, PPT2-EGFL8 targeting miR-423-5p plays an important role in hypoxia-induced peroxisome proliferator-activated receptor-beta/delta (PPARD)/angiopoietin-like 4 (ANGPTL4) signaling activation, especially the expression of the C-terminal ANGPTL4 fragment. Finally, ANGPTL4 significantly induces retinal vessel breakage in the inner limiting membrane and facilitates retinal vessel sprouting into the vitreous in the OIR mice. Thus, either new biomarkers or new therapeutic targets may be identified with translation of these findings.

Automated summary

Diabetic retinopathy (DR) is a common complication in patients with diabetes and the mechanisms involved in proliferative DR (PDR) causing blindness are not fully understood. miRNAs and long noncoding RNAs can accurately regulate target gene expression through competing endogenous RNAs. Through deep sequencing of extracellular vesicles (EVs) in the serum, aberrantly low expression of PPT2-EGFL8 and significantly increased level of miR-423-5p were found in individuals with PDR. PPT2-EGFL8 acts as a molecular sponge for miR-423-5p and inhibits hypoxia-induced proliferation of retinal endothelial cells. Egfl8-overexpression treatment reduces diabetes-related complications and pathological neovascularization in mouse models. The findings suggest the identification of potential biomarkers and therapeutic targets.