Hyperglycemia inhibition of endothelial miR-140-3p mediates angiogenic dysfunction in diabetes mellitus

Abstract

Objective: MicroRNAs (miRNAs) have emerged as promising regulators of diabetes mellitus (DM)-induced angiogenic dysfunction in endothelial cells (ECs), but information vis-à-vis the functional roles of distinct miRNAs remain surprisingly scarce. The current study was designed to elucidate the expression and function of miR-140-3p in diabetic ECs.

Methods: miR-140-3p expression was evaluated in DM mouse model and in human ECs using RT-qPCR, Northern blot and RNA fluorescent in situ hybridization. Effects of miR-140-3p manipulation on ECs function were evaluated using cell proliferation, migration and in vitro tube formation assay. Regulation of FOXK2 transcription by miR-140-3p was determined by luciferase reporter assay and site-directed mutagenesis.

Results: miR-140-3p expression was significantly down-regulated in high glucose-challenged ECs. Under normal conditions, miR-140-3p knockdown impaired endothelial proliferation and migration, and endothelial tube formation. Mechanistically, miR-140-3p exhibited its proangiogenic effects through directly inhibiting the expression of the forkhead transcription factor FOXK2. From a therapeutic standpoint, shRNA-mediated stable inhibition of FOXK2 effectively corrected miR-140-3p deficiency-induced impairment of ECs proliferation and in vitro angiogenesis.

Conclusion: Endothelial miR-140-3p positive regulates ECs function by directly targeting FOXK2 signaling. Deregulation of miR-140-3p/FOXK2 cascade by hyperglycemia thus serves as an important contributor to angiogenic dysfunction in DM.

Keywords: Angiogenesis; Diabetes mellitus (DM); FOXK2; Hyperglycemia; miRNA.