Cargando…
Conversion of hepatitis B virus relaxed circular to covalently closed circular DNA is supported in murine cells
BACKGROUND & AIMS: HBV has a narrow host restriction, with humans and chimpanzees representing the only known natural hosts. The molecular correlates of resistance in species that are commonly used in biomedical research, such as mice, are currently incompletely understood. Expression of human N...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9403495/ https://www.ncbi.nlm.nih.gov/pubmed/36035363 http://dx.doi.org/10.1016/j.jhepr.2022.100534 |
_version_ | 1784773390419099648 |
---|---|
author | Wei, Lei Cafiero, Thomas R. Tseng, Anna Gertje, Hans P. Berneshawi, Andrew Crossland, Nicholas A. Ploss, Alexander |
author_facet | Wei, Lei Cafiero, Thomas R. Tseng, Anna Gertje, Hans P. Berneshawi, Andrew Crossland, Nicholas A. Ploss, Alexander |
author_sort | Wei, Lei |
collection | PubMed |
description | BACKGROUND & AIMS: HBV has a narrow host restriction, with humans and chimpanzees representing the only known natural hosts. The molecular correlates of resistance in species that are commonly used in biomedical research, such as mice, are currently incompletely understood. Expression of human NTCP (hNTCP) in mouse hepatocytes enables HBV entry, but subsequently covalently closed circular (cccDNA) does not form in most murine cells. It is unknown if this blockade in cccDNA formation is due to deficiency in repair of relaxed circular DNA (rcDNA) to cccDNA. METHODS: Here, we deployed both in vivo and in vitro virological and biochemical approaches to investigate if murine cells contain a complete set of repair factors capable of converting HBV rcDNA to cccDNA. RESULTS: We demonstrate that HBV cccDNA does form in murine cell culture or in mice when recombinant rcDNA without a protein adduct is directly introduced into cells. We further show that the murine orthologues of core components in DNA lagging strand synthesis, required for the repair of rcDNA to cccDNA in human cells, can support this crucial step in the HBV life cycle. It is worth noting that recombinant HBV rcDNA substrates, either without a protein adduct or containing neutravidin to mimic HBV polymerase, were used in our study; it remains unclear if the HBV polymerase removal processes are the same in mouse and human cells. CONCLUSIONS: Collectively, our data suggest that the HBV life cycle is blocked post entry and likely before the repair stage in mouse cells, which yields critical insights that will aid in the construction of a mouse model with inbred susceptibility to HBV infection. LAY SUMMARY: Hepatitis B virus (HBV) is only known to infect humans and chimpanzees in nature. Mouse models are often used in modeling disease pathogenesis and preclinical research to assess the efficacy and safety of interventions before they are then tested in human participants. However, because mice are not susceptible to HBV infection it is difficult to accurately model human infection (and test potential treatments) in mouse models. Herein, we have shown that mice are able to perform a key step in the HBV life cycle, tightening the net around the possible reason why HBV can not efficiently infect and replicate in mice. |
format | Online Article Text |
id | pubmed-9403495 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-94034952022-08-26 Conversion of hepatitis B virus relaxed circular to covalently closed circular DNA is supported in murine cells Wei, Lei Cafiero, Thomas R. Tseng, Anna Gertje, Hans P. Berneshawi, Andrew Crossland, Nicholas A. Ploss, Alexander JHEP Rep Research Article BACKGROUND & AIMS: HBV has a narrow host restriction, with humans and chimpanzees representing the only known natural hosts. The molecular correlates of resistance in species that are commonly used in biomedical research, such as mice, are currently incompletely understood. Expression of human NTCP (hNTCP) in mouse hepatocytes enables HBV entry, but subsequently covalently closed circular (cccDNA) does not form in most murine cells. It is unknown if this blockade in cccDNA formation is due to deficiency in repair of relaxed circular DNA (rcDNA) to cccDNA. METHODS: Here, we deployed both in vivo and in vitro virological and biochemical approaches to investigate if murine cells contain a complete set of repair factors capable of converting HBV rcDNA to cccDNA. RESULTS: We demonstrate that HBV cccDNA does form in murine cell culture or in mice when recombinant rcDNA without a protein adduct is directly introduced into cells. We further show that the murine orthologues of core components in DNA lagging strand synthesis, required for the repair of rcDNA to cccDNA in human cells, can support this crucial step in the HBV life cycle. It is worth noting that recombinant HBV rcDNA substrates, either without a protein adduct or containing neutravidin to mimic HBV polymerase, were used in our study; it remains unclear if the HBV polymerase removal processes are the same in mouse and human cells. CONCLUSIONS: Collectively, our data suggest that the HBV life cycle is blocked post entry and likely before the repair stage in mouse cells, which yields critical insights that will aid in the construction of a mouse model with inbred susceptibility to HBV infection. LAY SUMMARY: Hepatitis B virus (HBV) is only known to infect humans and chimpanzees in nature. Mouse models are often used in modeling disease pathogenesis and preclinical research to assess the efficacy and safety of interventions before they are then tested in human participants. However, because mice are not susceptible to HBV infection it is difficult to accurately model human infection (and test potential treatments) in mouse models. Herein, we have shown that mice are able to perform a key step in the HBV life cycle, tightening the net around the possible reason why HBV can not efficiently infect and replicate in mice. Elsevier 2022-07-09 /pmc/articles/PMC9403495/ /pubmed/36035363 http://dx.doi.org/10.1016/j.jhepr.2022.100534 Text en © 2022 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Research Article Wei, Lei Cafiero, Thomas R. Tseng, Anna Gertje, Hans P. Berneshawi, Andrew Crossland, Nicholas A. Ploss, Alexander Conversion of hepatitis B virus relaxed circular to covalently closed circular DNA is supported in murine cells |
title | Conversion of hepatitis B virus relaxed circular to covalently closed circular DNA is supported in murine cells |
title_full | Conversion of hepatitis B virus relaxed circular to covalently closed circular DNA is supported in murine cells |
title_fullStr | Conversion of hepatitis B virus relaxed circular to covalently closed circular DNA is supported in murine cells |
title_full_unstemmed | Conversion of hepatitis B virus relaxed circular to covalently closed circular DNA is supported in murine cells |
title_short | Conversion of hepatitis B virus relaxed circular to covalently closed circular DNA is supported in murine cells |
title_sort | conversion of hepatitis b virus relaxed circular to covalently closed circular dna is supported in murine cells |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9403495/ https://www.ncbi.nlm.nih.gov/pubmed/36035363 http://dx.doi.org/10.1016/j.jhepr.2022.100534 |
work_keys_str_mv | AT weilei conversionofhepatitisbvirusrelaxedcirculartocovalentlyclosedcirculardnaissupportedinmurinecells AT cafierothomasr conversionofhepatitisbvirusrelaxedcirculartocovalentlyclosedcirculardnaissupportedinmurinecells AT tsenganna conversionofhepatitisbvirusrelaxedcirculartocovalentlyclosedcirculardnaissupportedinmurinecells AT gertjehansp conversionofhepatitisbvirusrelaxedcirculartocovalentlyclosedcirculardnaissupportedinmurinecells AT berneshawiandrew conversionofhepatitisbvirusrelaxedcirculartocovalentlyclosedcirculardnaissupportedinmurinecells AT crosslandnicholasa conversionofhepatitisbvirusrelaxedcirculartocovalentlyclosedcirculardnaissupportedinmurinecells AT plossalexander conversionofhepatitisbvirusrelaxedcirculartocovalentlyclosedcirculardnaissupportedinmurinecells |