Overexpression of miRNA-25-3p inhibits Notch1 signaling and TGF-β-induced collagen expression in hepatic stellate cells.
Berit GenzMiranda A ColemanKatharine M IrvineJamie Rose KutasovicMariska MirandaFrancis D GratteJanina E E Tirnitz-ParkerJohn K OlynykDiego A CalvopinaAnna WeisNicole CloonanHarley R RobinsonMichelle M HillFares Al-EjehGrant A RammPublished in: Scientific reports (2019)
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.
Keyphrases
- cell proliferation
- long non coding rna
- induced apoptosis
- long noncoding rna
- signaling pathway
- pi k akt
- poor prognosis
- transforming growth factor
- drug induced
- cell cycle arrest
- liver injury
- high glucose
- extracellular matrix
- epithelial mesenchymal transition
- young adults
- systemic sclerosis
- endoplasmic reticulum stress
- transcription factor
- single cell
- binding protein
- wound healing
- data analysis