Plerixafor and resatorvid inhibit hepatitis B virus in vitro by upregulating elongation factor Tu GTP-binding domain containing 2

Jinyuan Cai; Yuwen Li; Pingping Hu; Ruirui Xu; Hui Yuan; Wen Zhang; Tiantong Feng; Rui Liu; Wenting Li; Chuanlong Zhu

doi:10.3389/fcimb.2023.1118801

Plerixafor and resatorvid inhibit hepatitis B virus in vitro by upregulating elongation factor Tu GTP-binding domain containing 2

Front Cell Infect Microbiol. 2023 Feb 20:13:1118801. doi: 10.3389/fcimb.2023.1118801. eCollection 2023.

Authors

Jinyuan Cai^{1

2}, Yuwen Li³, Pingping Hu², Ruirui Xu², Hui Yuan², Wen Zhang², Tiantong Feng², Rui Liu⁴, Wenting Li⁴, Chuanlong Zhu^{1

4}

Affiliations

¹ 1Department of Infectious Disease, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China.
² Department of Infectious Disease, Zhongda Hospital, Southeast University, Nanjing, China.
³ Department of Pediatrics, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China.
⁴ Department of Infectious and Tropical Diseases, The Second Affiliated Hospital of Hainan Medical University, Hainan, China.

Abstract

Background: An increase in the demand for a functional cure has accelerated research on new methods of therapy for chronic hepatitis B, which is mainly focused on restoring antiviral immunity for controlling viral infections. Previously, we had described elongation factor Tu GTP-binding domain containing 2 (EFTUD2) as an innate immune regulator and suggested that it might be an antiviral target.

Methods: In this study, we generated the Epro-LUC-HepG2 cell model for screening compounds that target EFTUD2. Plerixafor and resatorvid were screened from 261 immunity and inflammation-related compounds due to their ability to highly upregulate EFTUD2. The effects of plerixafor and resatorvid on hepatitis B virus (HBV) were examined in HepAD38 cells and HBV-infected HepG2-NTCP cells.

Results: The dual-luciferase reporter assays showed that the EFTUD2 promoter hEFTUD2pro-0.5 kb had the strongest activity. In Epro-LUC-HepG2 cells, plerixafor and resatorvid significantly upregulated the activity of the EFTUD2 promoter and the expression of the gene and protein. In HepAD38 cells and HBV-infected HepG2-NTCP cells, treatment with plerixafor and resatorvid strongly inhibited HBsAg, HBV DNA, HBV RNAs, and cccDNA in a dose-dependent manner. Furthermore, the anti-HBV effect was enhanced when entecavir was administered along with either of the previous two compounds, and the effect could be blocked by knocking down EFTUD2.

Conclusion: We established a convenient model for screening compounds that target EFTUD2 and further identified plerixafor and resatorvid as novel HBV inhibitors in vitro. Our findings provided information on the development of a new class of anti-HBV agents that act on host factors rather than viral enzymes.

Keywords: EFTUD2; hepatitis B virus; plerixafor; resatorvid; small-molecule agents.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Antiviral Agents / pharmacology
Antiviral Agents / therapeutic use
DNA, Viral
Guanosine Triphosphate / pharmacology
Guanosine Triphosphate / therapeutic use
Hematopoietic Stem Cell Mobilization
Hep G2 Cells
Hepatitis B virus / physiology
Hepatitis B* / drug therapy
Heterocyclic Compounds* / pharmacology
Humans
Peptide Elongation Factor Tu / pharmacology
Peptide Elongation Factors / pharmacology
Ribonucleoprotein, U5 Small Nuclear / pharmacology
Virus Replication

Substances

ethyl 6-(N-(2-chloro-4-fluorophenyl)sulfamoyl)cyclohex-1-ene-1-carboxylate
Peptide Elongation Factor Tu
Heterocyclic Compounds
Antiviral Agents
Guanosine Triphosphate
DNA, Viral
EFTUD2 protein, human
Peptide Elongation Factors
Ribonucleoprotein, U5 Small Nuclear

Grants and funding

This work was supported by Science and Technology Plan of Hainan Province (Clinical Research Center) (LCYX202103 and LCYX202204), Hainan Province Science and Technology Special Fund (ZDYF2022SHFZ067), and the grants from Hainan Province Clinical Medical Center, the National Natural Science Foundation of China (No.81770591).