Overexpression of miRNA-25-3p inhibits Notch1 signaling and TGF-β-induced collagen expression in hepatic stellate cells

• Published in: Scientific reports 2019-06, Vol.9 (1), p.8541-8541, Article 8541
• Main Authors: Genz, Berit, Coleman, Miranda A., Irvine, Katharine M., Kutasovic, Jamie R., Miranda, Mariska, Gratte, Francis D., Tirnitz-Parker, Janina E. E., Olynyk, John. K., Calvopina, Diego A., Weis, Anna, Cloonan, Nicole, Robinson, Harley, Hill, Michelle M., Al-Ejeh, Fares, Ramm, Grant A.
• Format: Article
• Language: English
• Subjects: 13; 13/106; 13/95; 14; 14/32; 38; 38/109; 42; 42/41; 631/80/304; 692/4020/4021/1607/1605; 82; 96; Animals; Cell culture; Cell Line; Children; Collagen; Collagen - biosynthesis; Collagen - genetics; Cystic fibrosis; Extracellular matrix; Gene expression; Gene Expression Regulation; Hepatic Stellate Cells; Hepatocytes; Humanities and Social Sciences; Humans; Kinases; Liver; Liver diseases; Mice; MicroRNAs - genetics; MicroRNAs - metabolism; miRNA; multidisciplinary; Notch1 protein; Phosphorylation; Receptor, Notch1 - genetics; Receptor, Notch1 - metabolism; Science; Science (multidisciplinary); Signal transduction; Smad2 protein; Stellate cells; Transforming Growth Factor beta - genetics; Transforming Growth Factor beta - metabolism; Wnt protein; Wnt Signaling Pathway
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-019-44865-1

During chronic liver injury hepatic stellate cells (HSCs), the principal source of extracellular matrix in the fibrotic liver, transdifferentiate into pro-fibrotic myofibroblast-like cells - a process potentially regulated by microRNAs (miRNAs). Recently, we found serum miRNA-25-3p (miR-25) levels were upregulated in children with Cystic Fibrosis (CF) without liver disease, compared to children with CF-associated liver disease and healthy individuals. Here we examine the role of miR-25 in HSC biology. MiR-25 was detected in the human HSC cell line LX-2 and in primary murine HSCs, and increased with culture-induced activation. Transient overexpression of miR-25 inhibited TGF-β and its type 1 receptor (TGFBR1) mRNA expression, TGF-β-induced Smad2 phosphorylation and subsequent collagen1α1 induction in LX-2 cells. Pull-down experiments with biotinylated miR-25 revealed Notch signaling (co-)activators ADAM-17 and FKBP14 as miR-25 targets in HSCs. NanoString analysis confirmed miR-25 regulation of Notch- and Wnt-signaling pathways. Expression of Notch signaling pathway components and endogenous Notch1 signaling was downregulated in miR-25 overexpressing LX-2 cells, as were components of Wnt signaling such as Wnt5a. We propose that miR-25 acts as a negative feedback anti-fibrotic control during HSC activation by reducing the reactivity of HSCs to TGF-β-induced collagen expression and modulating the cross-talk between Notch, Wnt and TGF-β signaling.