Exosomal circTAOK1 contributes to diabetic kidney disease progression through regulating SMAD3 expression by sponging miR-520h.

Abstract

Diabetic nephropathy (DN) is a frequent diabetes complication with complex pathogenesis. Circular RNA (circRNA) circTAOK1 (also named circ_0003928) has been reported to be upregulated in high glucose (HG)-treated human umbilical vein endothelial cells. Also, exosomal circRNAs can exert significant roles in the pathology of various diseases. This study is designed to explore the role and mechanism of exosomal circTAOK1 on the glomerular mesangial cell (GMC) injury in DN. Exosomes were detected by a transmission electron microscope. The protein levels of CD9, CD63, proliferating cell nuclear antigen (PCNA), cyclinD1, α-SMA, fibronectin, E-cadherin, N-cadherin, and SMAD family member 3 (SMAD3) were examined by western blot assay. circTAOK1, microRNA-520h (miR-520h), and SMAD3 levels were detected by real-time quantitative polymerase chain reaction (RT-qPCR). Cell proliferation and cell cycle progression were detected by cell counting kit-8 (CCK-8), 5-ethynyl-2'-deoxyuridine (EdU), and flow cytometry assays. The binding relationship between miR-520h and circTAOK1 or SMAD3 was predicted by Starbase and then verified by a dual-luciferase reporter and RNA immunoprecipitation (RIP), RNA pull-down assays. CircTAOK1 expression was upregulated in the exosomes isolated from HG-treated glomerular epithelial cells (GEC). Moreover, GEC-circTAOK1-Exo could promote proliferation, fibrosis, and epithelial-mesenchymal transition (EMT) of glomerular mesangial cells (GMCs). Mechanically, circTAOK1 could regulate SMAD3 expression by sponging miR-520h, GEO-si-circTAOK1 Exo-induced miR-520h and repressed SMAD3 expression in GMC. GEC-circTAOK1-Exo could boost proliferation, fibrosis, and EMT of GMC through targeting the miR-520h/SMAD3 axis, providing new insights into the pathogenesis of DN.