E-cadherin Plays a Role in Hepatitis B Virus Entry Through Affecting Glycosylated Sodium-Taurocholate Cotransporting Polypeptide Distribution.

Qin Hu,Dandan Li,Pu Li,Liang Duan,Shengjun Yang,Feifei Zhang,Weixian Chen,Yuanyuan Ye,Lan Zhou,Bo Wang

doi:10.3389/fcimb.2020.00074

Abstract

Hepatitis B virus (HBV) infection is a major cause of chronic liver disease and hepatocellular carcinoma. Current antiviral therapy does not effectively eradicate HBV and further investigations into the mechanisms of viral infection are needed to enable the development of new therapeutic agents. The sodium-taurocholate cotransporting polypeptide (NTCP) has been identified as a functional receptor for HBV entry in liver cells. However, the NTCP receptor is not sufficient for entry and other membrane proteins contribute to modulate HBV entry. This study seeks to understand how the NTCP functions in HBV entry. Herein we show that knockdown of the cell-cell adhesion molecule, E-cadherin significantly reduced infection by HBV particles and entry by HBV pseudoparticles in infected liver cells and cell lines. The glycosylated NTCP localizes to the plasma membrane through interaction with E- cadherin, which increases interaction with the preS1 portion of the Large HBV surface antigen. Our study contributes novel insights that advance knowledge of HBV infection at the level of host cell binding and viral entry.

Highlights

Hepatitis B virus (HBV), a member of the Hepadnaviridae family, is a small, enveloped DNA virus (Glebe and Bremer, 2013) that infects human liver parenchymal cells
HepG2-NTCP, HepaRG, and Primary human hepatocytes (PHH) cells were transfected with 20 pmol of siRNA-NC, siRNA-E-cadherin, siRNA-NTCP or a combination of siRNA-E-cadherin and siRNA-NTCP before infection with 1 × 103 genome equivalents/cell of HBV produced from HepAD38 cells in 24-well plate
Silencing of E-cadherin and NTCP significantly reduced the level of HBV 3.5 kb mRNA in HepG2-NTCP, HepaRG, and PHH cell lines (Figure 1E); while silencing of both E-cadherin and NTCP in HepG2-NTCP and PHH cells served to further reduce the level of HBV 3.5 kb mRNA compared to that observed by either E-cadherin or NTCP separately

Summary

Introduction

Hepatitis B virus (HBV), a member of the Hepadnaviridae family, is a small, enveloped DNA virus (Glebe and Bremer, 2013) that infects human liver parenchymal cells. The sodium-taurocholate cotransporting polypeptide (NTCP), a bile acid transporter expressed at the basolateral membrane of human hepatocytes, has been identified as a functional receptor for HBV (Yan et al, 2012). Exogenous expression of NTCP in human hepatoma cell lines, such as HepG2 or Huh, confers susceptibility to HBV infection, and constitute effective cell culture models for examining HBV entry (Yan et al, 2012; Ni et al, 2014). The overexpression of NTCP in human or extrahepatic human cell lines, such as HeLa cells, or in mouse hepatocytes, is insufficient for productive HBV infection in these cells, suggesting that additional molecules are required for efficient HBV infection (Tong and Li, 2014)

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Results

Conclusion