Endothelial derived miRNA-9 mediated cardiac fibrosis in diabetes and its regulation by ZFAS1

Abstract

Diabetic cardiomyopathy (DCM) is one of the most prevalent causes of morbidity and mortality in diabetic patients. Hyperglycemia induces increased expression/deposition of extracellular matrix (ECM) proteins including fibronectin (FN) and collagen (Col) and plays an important role in fibrosis in diabetic cardiomyopathy (DCM). The roles of RNAs including microRNA (miRNA) and long non-coding RNAs (lncRNA) have begun to be understood in many conditions. In this study, we investigated the role of a specific miRNA, miR-9, and its interactions with lncRNA ZFAS1 in mediating fibrosis in DCM. Treatment with 25 mM glucose (HG) decreased miR-9 expression and increased expressions of ZFAS1, ECM proteins and inflammatory markers, compared to 5 mM glucose (NG) in the HCMECs by using qRT-PCR. Glucose-induced upregulation of ECM proteins can be prevented by ZFAS1 siRNA or miR-9 mimic transfection. Luciferase assay was confirmed miR-9 binding to FN 3’-UTR. miR-9 expression can be regulated by ZFAS1 through polycomb repressive complex 2 (PRC2) components using RNA immunoprecipitation (RIP) and chromatin immunoprecipitation (ChIP) assays. In the in vivo experiment, hyperglycemia-induced the ECM production can be prevented by the miR-9 overexpression in the fibrosis in DCM. These studies showed a novel glucose-induced molecular mechanism in which ZFAS1 participates in the transcriptional regulation of ECM protein production in diabetes through miR-9.

CircRNA_012164/MicroRNA-9-5p axis mediates cardiac fibrosis in diabetic cardiomyopathy

“…After showing the protective effects of miR-9 against DCM-related fibrotic changes in vivo , we continued the experiments in isolated mouse cardiac endothelial cells (MCECs) to gain mechanistic insights specific to endothelial cells and endothelial dysfunction. In response to treatment with 30 mM D-glucose, MCECs showed significant elevation in the levels of circRNA_012164 as well as a reduction in miR9 levels corresponding to previous literature and our in vivo studies(44) (Fig 4A, B). No such changes were observed with osmotic control using mannitol (not shown).…”

MicroRNA 9 Is a Regulator of Endothelial to Mesenchymal Transition in Diabetic Retinopathy

“…Further investigation into the mechanistic link between high glucose and miR-9 suppression will be required to fully elucidate the glucose-miR-9-endMT pathway in the retina. Our laboratory has recently demonstrated a link between glucose-induced upregulation of lncRNA ZFAS1 and the suppression of miR-9 in cardiac fibrosis, 67 but this relation needs to be validated in DR in future experiments.…”

Endothelial to mesenchymal cell transition in diabetic retinopathy: targets and therapeutics

“…Short interfering RNAs (siRNAs) targeting pro-EndMT genes and lncRNAs that mimic anti-EndMT microRNAs shown promise in preventing EndMT. Experimental silencing of lncRNAs ZFAS1 and MALAT1, as well as induction of various miRNAs, have proven to be effective in suppressing hyperglycemia-induced EndMT through different molecular pathways (76,(94)(95)(96)(97)(98). Moreover, synthetic lncRNAs have emerged as a potential strategy to suppress glucose-induced EndMT (99).…”