HBV Pre-S1-Derived Myristoylated Peptide (Myr47): Identification of the Inhibitory Activity on the Cellular Uptake of Lipid Nanoparticles.

Masaya Nanahara,Ya-Ting Chang,Masaharu Somiya,Shun’Ichi Kuroda

doi:10.3390/v13050929

Abstract

The Myr47 lipopeptide, consisting of hepatitis B virus (HBV) pre-S1 domain (myristoylated 2–48 peptide), is an effective commercialized anti-HBV drug that prevents the interaction of HBV with sodium taurocholate cotransporting polypeptide (NTCP) on human hepatocytes, an activity which requires both N-myristoylation residue and specific amino acid sequences. We recently reported that Myr47 reduces the cellular uptake of HBV surface antigen (HBsAg, subviral particle of HBV) in the absence of NTCP expression. In this study, we analyzed how Myr47 reduces the cellular uptake of lipid nanoparticles (including liposomes (LPs) and HBsAg) without NTCP expression. By using Myr47 mutants lacking the HBV infection inhibitory activity, they could reduce the cellular uptake of LPs in an N-myristoylation-dependent manner and an amino acid sequence-independent manner, not only in human liver-derived cells but also in human non-liver-derived cells. Moreover, Myr47 and its mutants could reduce the interaction of LPs with apolipoprotein E3 (ApoE3) in an N-myristoylation-dependent manner regardless of their amino acid sequences. From these results, lipopeptides are generally anchored by inserting their myristoyl residue into the lipid bilayer and can inhibit the interaction of LPs/HBsAg with apolipoprotein, thereby reducing the cellular uptake of LPs/HBsAg. Similarly, Myr47 would interact with HBV, inhibiting the uptake of HBV into human hepatic cells, while the inhibitory effect of Myr47 may be secondary to its ability to protect against HBV infection.

Highlights

Hepatitis B virus (HBV) is a DNA virus infecting the human liver hepatocyte, and causes serious liver diseases such as hepatitis, cirrhosis, and liver cancer. HBV was discovered nearly half a century ago, the curative agent has not yet been developed [1], and only HB vaccine can prevent the horizontal and vertical transmission of HBV
Since the cellular uptake of HBsAg by Hep G2 cells was efficiently inhibited by Myr47 [11], we speculated that this inhibitory activity was not directly related to HBV envelope proteins, but to the lipid nanoparticle structure
The inhibitory activity depends on the N-myristoyl residue but not on the amino acid sequence of lipopeptides

Summary

Introduction

Hepatitis B virus (HBV) is a DNA virus infecting the human liver hepatocyte, and causes serious liver diseases such as hepatitis, cirrhosis, and liver cancer. HBV was discovered nearly half a century ago, the curative agent has not yet been developed [1], and only HB vaccine can prevent the horizontal and vertical transmission of HBV. Hepatitis B virus (HBV) is a DNA virus infecting the human liver hepatocyte, and causes serious liver diseases such as hepatitis, cirrhosis, and liver cancer. S, M, and L protein that share the same C-terminal sequence [2]. The pre-S1 region of the L protein is essential for specific recognition of human hepatocytes [3,4,5]. An essential region for hepatocyte recognition was identified as 9–18 aa of the pre-S1 region [9]. In 2012, the sodium taurocholate cotransporting polypeptide (NTCP) that is expressed in hepatocyte was identified as a critical cellular receptor for HBV early infection (cell entry step) [10]

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Conclusion