MiR-16-5p suppresses myofibroblast activation in systemic sclerosis by inhibiting NOTCH signaling

Abstract

Systemic sclerosis (SSc) is a prototypic fibrotic disease characterized by localized or diffuse skin thickening and fibrosis. Tissue fibrosis is driven by myofibroblasts, and factors affecting myofibroblast activation may also be involved in the development of SSc. In this study, we examined molecular mechanisms underlying SSc by focusing on myofibroblast activation processes. Bioinformatics analysis conducted to identify differentially expressed miRNAs (DEMs) and genes (DEGs) revealed that microRNA-16-5p (miR-16-5p) was downregulated and NOTCH2 was upregulated in SSc patients. In vitro experiments confirmed that miR-16-5p was able to bind directly to NOTCH2 and inhibit myofibroblast activation. Moreover, miR-16-5p-dependent inhibition of NOTCH2 decreased collagen and α-SMA expression. MiR-16-5p downregulation and NOTCH2 upregulation was also confirmed in vivo in SSc patients, and NOTCH2 activation promoted fibrosis progression in vitro . These results indicate that miR-16-5p suppresses myofibroblast activation by suppressing NOTCH signaling.

The Molecular Mechanisms and Function of miR-15a/16 Dysregulation in Fibrotic Diseases

“…Manesh et al [56] found that miR-16 may affect the important pathogenic events of systemic sclerosis by targeting the nuclear proteins survivin and p53 that control cell cycle and apoptosis, inhibiting endothelial cell proliferation and promoting apoptosis. Similarly, it has been found that miR-16-5p can directly bind Smad3 [22] and Notch2 [26] to inhibit the activation and phenotypic transformation of fibroblasts and reduce the synthesis of collagen and other ECM and α-Smooth muscle actin(α-SMA) expressions, indicating that it has a direct anti-fibrotic effect.…”

“…In addition, miR-15a and miR-16 can also affect other signaling pathways related to ECM formation. Yao et al [26] found that miR-16-5p inhibited the activation of the Notch signaling pathway by targeting Notch2, thereby increasing the expression of MMP-1 and MMP-8 in human skin fibroblasts and promoting the degradation of collagen and other ECM components. Chen et al [27] found that the overexpression of miR-15a inhibited Yesassociated protein (YAP1), a key effector molecule in the Hippo pathway, and affected the activation of the downstream signal YAP1/TEAD/Twist, thereby inhibiting the synthesis of collagen and other ECM components in lung fibroblasts.…”

Notch signaling pathway: a comprehensive prognostic and gene expression profile analysis in breast cancer

“…This miRNA has been shown to suppress breast cancer proliferation [ 125 ]. Another study demonstrated that miR-16-5p has an inhibiting role in the Notch signaling pathway [ 126 ]. Therefore, it could be a diagnostic and prognostic potential biomarker in breast cancer.…”

An update on epigenetic regulation in autoimmune diseases

“…MicroRNAs also exert epigenetic regulation on fibroblasts. Low level of miR-16–5p activates fibrosis by positively regulating the NOTCH signaling [ 144 ]. In addition, miR33a-3p can downregulate level of DKK-1, an antagonist of Wnt pathway, and promote the fibrosis [ 145 ].…”