The feasibility of establishing a hamster model for HBV infection: in vitro evidence
ABSTRACT Chronic hepatitis B virus (HBV) infection remains a significant public health burden with no cure currently available. The research to cure HBV has long been hampered by the lack of immunocompetent small animal models capable of supporting HBV infection. Here, we set out to explore the feas...
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American Society for Microbiology
2024-11-01
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| Online Access: | https://journals.asm.org/doi/10.1128/mbio.02615-24 |
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| author | Hu Zhang Yanan Liu Cheng-Der Liu Zhongde Wang Haitao Guo |
| author_facet | Hu Zhang Yanan Liu Cheng-Der Liu Zhongde Wang Haitao Guo |
| author_sort | Hu Zhang |
| collection | DOAJ |
| description | ABSTRACT Chronic hepatitis B virus (HBV) infection remains a significant public health burden with no cure currently available. The research to cure HBV has long been hampered by the lack of immunocompetent small animal models capable of supporting HBV infection. Here, we set out to explore the feasibility of the golden Syrian hamster as an immunocompetent small rodent model for HBV infection. We first started with in vitro assessments of the HBV replication cycle in primary hamster hepatocytes (PHaHs) by adenoviral HBV (Ad-HBV) transduction. Our results demonstrated that PHaHs support HBV reverse transcription and subsequent cccDNA formation via the intracellular recycling pathway. Next, with luciferase reporter assays, we confirmed that PHaHs support the activities of all HBV major promoters. Then, we transduced PHaHs with an adenoviral vector expressing HBV receptor human Na+/taurocholate cotransporting polypeptide NTCP (Ad-huNTCP), followed by HBV inoculation. While the untransduced PHaHs did not support HBV infection, Ad-huNTCP-transduced PHaHs supported de novo cccDNA formation, viral mRNA transcription, and expression of viral antigens. We then humanized the amino acid (aa) residues of hamster NTCP (haNTCP) critical for HBV entry, aa84-87 and aa157-165, and transfected HepG2 cells with constructs expressing wild-type haNTCP and humanized-haNTCP, H84R/P87N and H84R/P87N/G157K/M160V/M165L, respectively, followed by HBV inoculation. The results showed that the humanization of H84R/P87N alone was sufficient to support HBV infection at a level comparable to that supported by huNTCP. Taken together, the above in vitro evidence supports the future direction of humanizing haNTCP for HBV infection in vivo.IMPORTANCEOne of the biggest challenges in developing an HBV cure is the lack of immunocompetent animal models susceptible to HBV infection. Developing such models in mice has been unsuccessful due to the absence of a functional HBV receptor, human NTCP (huNTCP), and the defect in supporting viral cccDNA formation. In search of alternative models, we report herein multiple lines of in vitro evidence for developing a golden Syrian hamster model for HBV infection. We demonstrate that the primary hamster hepatocytes (PHaHs) support HBV replication, transcription, and cccDNA formation, and PHaHs are susceptible to de novo HBV infection in the presence of huNTCP. Furthermore, expressing hamster NTCP with two humanized residues critical for HBV entry renders HepG2 cells permissive to HBV infection. Thus, our work lays a solid foundation for establishing a gene-edited hamster model that expresses humanized NTCP for HBV infection in vivo. |
| format | Article |
| id | doaj-art-9ec8e93bdbab45df9153817ec895762b |
| institution | OA Journals |
| issn | 2150-7511 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | American Society for Microbiology |
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| spelling | doaj-art-9ec8e93bdbab45df9153817ec895762b2025-08-20T02:14:38ZengAmerican Society for MicrobiologymBio2150-75112024-11-01151110.1128/mbio.02615-24The feasibility of establishing a hamster model for HBV infection: in vitro evidenceHu Zhang0Yanan Liu1Cheng-Der Liu2Zhongde Wang3Haitao Guo4Cancer Virology Program, UPMC Hillman Cancer Center, University of Pittsburgh, School of Medicine, Pittsburgh, Pennsylvania, USADepartment of Animal, Dairy and Veterinary Sciences, Utah State University, Logan, Utah, USACancer Virology Program, UPMC Hillman Cancer Center, University of Pittsburgh, School of Medicine, Pittsburgh, Pennsylvania, USADepartment of Animal, Dairy and Veterinary Sciences, Utah State University, Logan, Utah, USACancer Virology Program, UPMC Hillman Cancer Center, University of Pittsburgh, School of Medicine, Pittsburgh, Pennsylvania, USAABSTRACT Chronic hepatitis B virus (HBV) infection remains a significant public health burden with no cure currently available. The research to cure HBV has long been hampered by the lack of immunocompetent small animal models capable of supporting HBV infection. Here, we set out to explore the feasibility of the golden Syrian hamster as an immunocompetent small rodent model for HBV infection. We first started with in vitro assessments of the HBV replication cycle in primary hamster hepatocytes (PHaHs) by adenoviral HBV (Ad-HBV) transduction. Our results demonstrated that PHaHs support HBV reverse transcription and subsequent cccDNA formation via the intracellular recycling pathway. Next, with luciferase reporter assays, we confirmed that PHaHs support the activities of all HBV major promoters. Then, we transduced PHaHs with an adenoviral vector expressing HBV receptor human Na+/taurocholate cotransporting polypeptide NTCP (Ad-huNTCP), followed by HBV inoculation. While the untransduced PHaHs did not support HBV infection, Ad-huNTCP-transduced PHaHs supported de novo cccDNA formation, viral mRNA transcription, and expression of viral antigens. We then humanized the amino acid (aa) residues of hamster NTCP (haNTCP) critical for HBV entry, aa84-87 and aa157-165, and transfected HepG2 cells with constructs expressing wild-type haNTCP and humanized-haNTCP, H84R/P87N and H84R/P87N/G157K/M160V/M165L, respectively, followed by HBV inoculation. The results showed that the humanization of H84R/P87N alone was sufficient to support HBV infection at a level comparable to that supported by huNTCP. Taken together, the above in vitro evidence supports the future direction of humanizing haNTCP for HBV infection in vivo.IMPORTANCEOne of the biggest challenges in developing an HBV cure is the lack of immunocompetent animal models susceptible to HBV infection. Developing such models in mice has been unsuccessful due to the absence of a functional HBV receptor, human NTCP (huNTCP), and the defect in supporting viral cccDNA formation. In search of alternative models, we report herein multiple lines of in vitro evidence for developing a golden Syrian hamster model for HBV infection. We demonstrate that the primary hamster hepatocytes (PHaHs) support HBV replication, transcription, and cccDNA formation, and PHaHs are susceptible to de novo HBV infection in the presence of huNTCP. Furthermore, expressing hamster NTCP with two humanized residues critical for HBV entry renders HepG2 cells permissive to HBV infection. Thus, our work lays a solid foundation for establishing a gene-edited hamster model that expresses humanized NTCP for HBV infection in vivo.https://journals.asm.org/doi/10.1128/mbio.02615-24hepatitis B virushamster hepatocytesNTCPcccDNA |
| spellingShingle | Hu Zhang Yanan Liu Cheng-Der Liu Zhongde Wang Haitao Guo The feasibility of establishing a hamster model for HBV infection: in vitro evidence mBio hepatitis B virus hamster hepatocytes NTCP cccDNA |
| title | The feasibility of establishing a hamster model for HBV infection: in vitro evidence |
| title_full | The feasibility of establishing a hamster model for HBV infection: in vitro evidence |
| title_fullStr | The feasibility of establishing a hamster model for HBV infection: in vitro evidence |
| title_full_unstemmed | The feasibility of establishing a hamster model for HBV infection: in vitro evidence |
| title_short | The feasibility of establishing a hamster model for HBV infection: in vitro evidence |
| title_sort | feasibility of establishing a hamster model for hbv infection in vitro evidence |
| topic | hepatitis B virus hamster hepatocytes NTCP cccDNA |
| url | https://journals.asm.org/doi/10.1128/mbio.02615-24 |
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