miRNA‐200c‐3p promotes endothelial to mesenchymal transition and neointimal hyperplasia in artery bypass grafts

Abstract

Increasing evidence has suggested a critical role for endothelial‐to‐mesenchymal transition (EndoMT) in a variety of pathological conditions. MicroRNA‐200c‐3p (miR‐200c‐3p) has been implicated in epithelial‐to‐mesenchymal transition. However, the functional role of miR‐200c‐3p in EndoMT and neointimal hyperplasia in artery bypass grafts remains largely unknown. Here we demonstrated a critical role for miR‐200c‐3p in EndoMT. Proteomics and luciferase activity assays revealed that fermitin family member 2 (FERM2) is the functional target of miR‐200c‐3p during EndoMT. FERMT2 gene inactivation recapitulates the effect of miR‐200c‐3p overexpression on EndoMT, and the inhibitory effect of miR‐200c‐3p inhibition on EndoMT was reversed by FERMT2 knockdown. Further mechanistic studies revealed that FERM2 suppresses smooth muscle gene expression by preventing serum response factor nuclear translocation and preventing endothelial mRNA decay by interacting with Y‐box binding protein 1. In a model of aortic grafting using endothelial lineage tracing, we observed that miR‐200c‐3p expression was dramatically up‐regulated, and that EndoMT contributed to neointimal hyperplasia in grafted arteries. MiR‐200c‐3p inhibition in grafted arteries significantly up‐regulated FERM2 gene expression, thereby preventing EndoMT and reducing neointimal formation. Importantly, we found a high level of EndoMT in human femoral arteries with atherosclerotic lesions, and that miR‐200c‐3p expression was significantly increased, while FERMT2 expression levels were dramatically decreased in diseased human arteries. Collectively, we have documented an unexpected role for miR‐200c‐3p in EndoMT and neointimal hyperplasia in grafted arteries. Our findings offer a novel therapeutic opportunity for treating vascular diseases by specifically targeting the miR‐200c‐3p/FERM2 regulatory axis. © 2020 The Authors. The Journal of Pathology published by John Wiley & Sons, Ltd. on behalf of The Pathological Society of Great Britain and Ireland.

Neutrophil elastase promotes neointimal hyperplasia by targeting toll‐like receptor 4 (TLR4)–NF‐κB signalling

“…WT and neutrophil elastase_KO VSMCs (0.75 × 10 4 per well) were cultured in 96‐well plates overnight, followed by serum starvation for 48 h. Starved VSMCs were incubated with 10‐μM H 2 O 2 for additional 24 h to induce apoptosis. Cell viability was evaluated using Cell Counting Kit‐8 (Sigma/Merck, 96992‐500TESTS‐F) according to the manufacturer's instructions as described previously (Chen et al, 2021). The absorbance of the samples was measured by a microplate reader at 450 nm (OD 450 ), and relative absorbance ( A 450 nm ) values representing cell viability were compared between treatments with control sample set as 1.0.…”

“…Proteomic studies and data analysis were conducted as described in our previous studies (Afzal et al, 2016; Chen et al, 2021; Xiao et al, 2014). Briefly, serum‐starved neutrophil elastase_KO VSMCs treated with vehicle or 100‐ng·ml −1 active human neutrophil elastase (ab91099) were directly lysed in a urea‐based lysis buffer, and proteins were digested using trypsin.…”

EndMT Regulation by Small RNAs in Diabetes-Associated Fibrotic Conditions: Potential Link With Oxidative Stress

“…Moreover, miR-200b overexpression also induces the down-regulation of p300, a transcription coactivator known to contribute to cardiac fibrosis and hypertrophy via TGF-β/SMAD ( Bugyei-Twum et al, 2014 ; Feng et al, 2016 ). Although the inhibitory role of the whole miR-200 family is well established, both in EMT ( Korpal and Kang, 2008 ; Korpal et al, 2008 ) and EndMT ( Feng et al, 2016 ; Zhang et al, 2017 ), unexpectedly a recent study shown that miR-200c-3p exerted the opposite effect, being able to promote EndMT and aortic graft remodeling both in vivo and in vitro ( Chen et al, 2021 ). Finally, a further TGF-β/SMAD pathway-mediated regulatory mechanism involves miR-451 whose effects on EndMT are AMPK-dependent.…”

MiR-200c-3p promotes ox-LDL-induced endothelial to mesenchymal transition in human umbilical vein endothelial cells through SMAD7/YAP pathway

“…For example, the EndMT process that relies on miR-200 family is a reasonable biological mechanism that makes the bicuspid aortic valve more prone to aneurysms [22]. A previous study has confirmed that miR-200c-3p is highly expressed in human femoral arteries with atherosclerotic lesions [23]. The miR-200c-3p expression is also increased in the NaF-treated HUVECs, and NaF promotes apoptosis of HUVECs by regulating miR-200c-3p/Fas pathway [24].…”