Mesenchymal stem cell-originated exosomal circDIDO1 suppresses hepatic stellate cell activation by miR-141-3p/PTEN/AKT pathway in human liver fibrosis

Abstract

Liver fibrosis is a common pathologic stage of the development of liver failure. It has showed that exosomes loaded with therapeutic circRNAs can be manufactured in bulk by exosome secreted cells in vitro , thus enabling personalized treatment. This study aimed to investigate the role of exosome-based delivery of circDIDO1 in liver fibrosis. Levels of genes and proteins were examined by qRT-PCR and Western blot. Cell proliferation, apoptosis, and cell cycle were analyzed by using cell counting kit-8 (CCK-8) assay, EdU assay, and flow cytometry, respectively. The binding between circDIDO1 and miR-141-3p was confirmed by dual-luciferase reporter, RNA pull-down and RIP assays. Exosomes were isolated by ultracentrifugation, and qualified by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA) and Western blot. CircDIDO1 overexpression or miR-141-3p inhibition suppressed the proliferation, reduced pro-fibrotic markers, and induced apoptosis as well as cell cycle arrest in hepatic stellate cells (HSCs) by blocking PTEN/AKT pathway. Mechanistically, circDIDO1 acted as an endogenous sponge for miR-141-3p, further rescue experiments showed that circDIDO1 suppressed HSC activation by targeting miR-141-3p. Extracellular circDIDO1 could be incorporated into exosomes isolated from mesenchymal stem cells (MSCs), and transmitted to HSCs to restrain HSC activation. Clinically, low levels of serum circDIDO1 in exosome were correlated with liver failure, and serum exosomal circDIDO1 had a well diagnostic value for liver fibrosis in liver failure patients. Transfer of circDIDO1 mediated by MSC-isolated exosomes suppressed HSC activation through the miR-141-3p/PTEN/AKT pathway, gaining a new insight into the prevention of liver fibrosis in liver failure patients.

Structural and Temporal Dynamics of Mesenchymal Stem Cells in Liver Diseases From 2001 to 2021: A Bibliometric Analysis

“…Interestingly, a variety of microRNAs in the extracellular vesicles of MSCs show anti-hepatoma effects, such as miR−199a−3p, miR-199a, miR-15a and miR-375 ( 136 – 139 ). In addition, long non-coding RNAs (lncRNA) and circular RNAs (circRNA), as components of non-coding RNAs, also play an irreplaceable role in the underlying mechanism of MSCs in the treatment of liver diseases ( 140 , 141 ).…”

Stem cells for treatment of liver fibrosis/cirrhosis: clinical progress and therapeutic potential

“…Amnion-derived MSC (AMSC) was displayed to repress hepatic stellate cells activation by inhibiting earlier steps of the LPS/TLR4 pathways but not downstream NF-κB transcriptional activity [ 126 ]. MSC-originated exosomal circDIDO1 sponged miR-143-3p in hepatic stellate cells, causing cell cycle arrest, suppression, and apoptosis through promoting PTEN and repressing the ratio of p-AKT/AKT [ 127 ]. More, BMMSC transplantation significantly reduced the NOXs which are vital intermediates in the motivation of hepatic stellate cells through decreasing phosphorylation of p47phox, a ligand of NOX [ 128 ].…”

TGFβ1-Pretreated Exosomes of Wharton Jelly Mesenchymal Stem Cell as a Therapeutic Strategy for Improving Liver Fibrosis

“…An ECL system was used to visualize the blots. The extracted exosomes (50 µg/mL) (34,35) were added to the plates to treat HSCs, while control cells remained untreated.…”