MicroRNA-dependent cross-talk between VEGF and HIF1α in the diabetic retina

Abstract

Both HIF1α (hypoxia-inducible factor alpha) and VEGF (vascular endothelial growth factor) are implicated in the pathogenesis of diabetic retinopathy (DR). Competitive endogenous RNAs (ceRNAs) are messenger RNA (mRNA) molecules that affect each other expression through competition for their shared microRNAs (miRNA). However, little is known about the role of ceRNAs in DR. We assess whether the expression of HIF1α and VEGF in DR is interdependent through sequestration of common miRNAs. We used bioinformatics to identify potential miRNAs that affect both genes and validated the interdependence of the genes by silencing or overexpression of the genes and assessed the luciferase-HIF1α 3'UTR activity. We found that HIF1α and VEGF are targeted by 12 common miRNAs. Silencing either HIF1α or VEGF increased the availabilities of the shared miRNAs, therefore suppressed the luciferase-HIF1α 3'UTR activity, whereas over-expressing HIF1α or VEGF increased the luciferase activity. HIF1α was co-expressed with VEGF in-vivo and in-vitro in DR models. Silencing HIF1α transcripts resulted in a significant reduction in VEGF protein levels and vice versa. This interdependence was miRNA- and 3'UTR-dependent, as silencing Dicer abolished the interdependence. Over-expression of a common miRNA (miR-106a) significantly reduced the expression of HIF1α and VEGF and prevented high glucose-induced increased permeability. There is a cross-talk between HIF1α and VEGF through interactions with their common miRNAs. miRNA based therapy can affect the expression of both HIF1α and VEGF and may represent a therapeutic potential for the treatment of DR.

Keywords: 3′ untranslated region; 3′UTR; CeRNA; DKK1; DM; DR; Diabetic retinopathy; Dickkopf-1; HG; HIF1α; ICAM-1; MRE; MicroRNA; NG; PTEN; RPE; STZ; VEGF; ceRNA; competing endogenous RNA; diabetes mellitus; diabetic retinopathy; high glucose; hypoxia-inducible factor 1-alpha; intercellular adhesion molecule-1; mRNA; messenger RNA; miRNA; microRNA; microRNA response element; normal glucose; phosphatase and tensin homologue on chromosome 10; retinal pigment epithelium; siRNA; small interfering RNA; streptozotocin; vascular endothelial growth factor.