Human Liver Stem Cell-Derived Extracellular Vesicles Target Hepatic Stellate Cells and Attenuate Their Pro-fibrotic Phenotype.

Giulia Chiabotto,Stefania Bruno,Giovanni Camussi,Marta Tapparo,Elena Ceccotti

doi:10.3389/fcell.2021.777462

Giulia Chiabotto, Stefania Bruno + Show 3 more

Open Access

https://doi.org/10.3389/fcell.2021.777462

Copy DOI

Abstract

Liver fibrosis occurs in response to chronic liver injury and is characterized by an excessive deposition of extracellular matrix. Activated hepatic stellate cells are primarily responsible for this process. A possible strategy to counteract the development of hepatic fibrosis could be the reversion of the activated phenotype of hepatic stellate cells. Extracellular vesicles (EVs) are nanosized membrane vesicles involved in intercellular communication. Our previous studies have demonstrated that EVs derived from human liver stem cells (HLSCs), a multipotent population of adult stem cells of the liver with mesenchymal-like phenotype, exert in vivo anti-fibrotic activity in the liver. However, the mechanism of action of these EVs remains to be determined. We set up an in vitro model of hepatic fibrosis using a human hepatic stellate cell line (LX-2) activated by transforming growth factor-beta 1 (TGF-β1). Then, we investigated the effect of EVs obtained from HLSCs and from human bone marrow-derived mesenchymal stromal cells (MSCs) on activated LX-2. The incubation of activated LX-2 with HLSC-EVs reduced the expression level of alpha-smooth muscle actin (α-SMA). Conversely, MSC-derived EVs induced an increase in the expression of pro-fibrotic markers in activated LX-2. The analysis of the RNA cargo of HLSC-EVs revealed the presence of several miRNAs involved in the regulation of fibrosis and inflammation. Predictive target analysis indicated that several microRNAs (miRNAs) contained into HLSC-EVs could possibly target pro-fibrotic transcripts. In particular, we demonstrated that HLSC-EVs shuttled miR-146a-5p and that treatment with HLSC-EVs increased miR-146a-5p expression in LX-2. In conclusion, this study demonstrates that HLSC-EVs can attenuate the activated phenotype of hepatic stellate cells and that their biological effect may be mediated by the delivery of anti-fibrotic miRNAs, such as miR-146a-5p.

Highlights

Hepatic fibrosis is a reversible wound-healing response that originates from a chronic liver injury
Both extracellular vesicle (EV) from human liver stem cell (HLSC) and mesenchymal stromal cell (MSC) showed the expression of the classical exosomal markers CD9, CD63, CD81, and the mesenchymal markers CD29, CD44, and CD105
Transmission electron microscopy analysis showed that HLSC-EVs and MSC-EVs retain a homogeneous pattern of nano-size membrane vesicles (Figures 1D,E)

Summary

Introduction

Hepatic fibrosis is a reversible wound-healing response that originates from a chronic liver injury. The development of liver fibrosis is mainly due to an increased production and deposition of extracellular matrix (ECM) components along with an imbalance in ECM degradation, which compromises the normal morphology and function of the liver (Schuppan et al, 2018; Parola and Pinzani, 2019; Roehlen et al, 2020). During the early phases of liver fibrosis the activation of the hepatic stellate cell (HSC) from a quiescent vitamin A-storing cell to a myofibroblastlike cell has been shown (Bataller and Brenner, 2005). A possible strategy to counteract the development of hepatic fibrosis could be the reversion of the activated state of HSCs

Objectives

Methods

Results

Conclusion