Endothelial cell‐derived matrix promotes the metabolic functional maturation of hepatocyte via integrin‐Src signalling

Xinyue Guo,Minghui Ma,Xin Lu,Weihong Li,Haiyan Zhang

doi:10.1111/jcmm.13195

Abstract

The extracellular matrix (ECM) microenvironment is involved in the regulation of hepatocyte phenotype and function. Recently, the cell‐derived extracellular matrix has been proposed to represent the bioactive and biocompatible materials of the native ECM. Here, we show that the endothelial cell‐derived matrix (EC matrix) promotes the metabolic maturation of human adipose stem cell‐derived hepatocyte‐like cells (hASC‐HLCs) through the activation of the transcription factor forkhead box protein A2 (FOXA2) and the nuclear receptors hepatocyte nuclear factor 4 alpha (HNF4α) and pregnane X receptor (PXR). Reducing the fibronectin content in the EC matrix or silencing the expression of α5 integrin in the hASC‐HLCs inhibited the effect of the EC matrix on Src phosphorylation and hepatocyte maturation. The inhibition of Src phosphorylation using the inhibitor PP2 or silencing the expression of Src in hASC‐HLCs also attenuated the up‐regulation of the metabolic function of hASC‐HLCs in a nuclear receptor‐dependent manner. These data elucidate integrin‐Src signalling linking the extrinsic EC matrix signals and metabolic functional maturation of hepatocyte. This study provides a model for studying the interaction between hepatocytes and non‐parenchymal cell‐derived matrix.

Highlights

The severe shortage of liver donors for human primary adult hepatocyte has prompted in vitro efforts to produce the functional hepatocyte-like cells (HLCs) from stem cells [1, 2]
The results showed that the mRNA levels of forkhead box protein A2 (FOXA2), hepatocyte nuclear factor 4 alpha (HNF4a) and pregnane X receptor (PXR) in the Human adipose stem cells (hASCs)-HLCs cultured on the FN-depleted EC matrix showed expected 37%, 59% and 73% decreases compared to the cells cultured on the control EC matrix
We provided evidence that the EC matrix modulated the expression of FOXA2, HNF4a and PXR and regulated the expression of several of their downstream functional targets, including CYP450 enzymes and transporters, promoting the metabolic functional maturation of a 2017 The Authors

Summary

Introduction

The severe shortage of liver donors for human primary adult hepatocyte has prompted in vitro efforts to produce the functional hepatocyte-like cells (HLCs) from stem cells [1, 2]. HLCs have been differentiated from hASCs in vitro by toning soluble maintenance factors or microRNA to direct cell fate [5,6,7,8]. The ECM proteins produced provide an adhesive substrate for integrins and a platform for the transduction of intracellular signalling events that regulate a host of cell functions and that play an important role in maintaining the differentiated phenotype [12, 14, 15]

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Results

Conclusion