{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2023,3,21]],"date-time":"2023-03-21T05:02:25Z","timestamp":1679374945817},"reference-count":37,"publisher":"Institute of Physiology of the Czech Academy of Sciences","issue":"1","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Physiol Res"],"abstract":"<jats:p>Chronic hepatitis B (CHB) is caused by the Hepatitis B virus (HBV) and affects millions of people worldwide. Developing an effective CHB therapy requires using in vivo screening methods, such as mouse models reflecting CHB based on hydrodynamic delivery of plasmid vectors containing a replication-competent HBV genome. However, long-term expression of HBV proteins is accompanied by production of progeny virions, thereby requiring a Biosafety Level (BSL) 3 animal facility. In the present study, we introduced a point mutation in the START codon of the HBV polymerase to develop a mouse model reflecting chronic hepatitis B infection without formation of viral progeny. We induced the mouse model by hydrodynamic injection of adeno-associated virus plasmid vector (pAAV) and minicircle plasmid (pMC) constructs into C57Bl\/6 and C3H\/HeN mouse strains, monitoring HBV antigens and antibodies in blood by enzyme-linked immunosorbent assay and analyzing liver expression of HBV core antigen by immunohistology. Persisting expression of viral antigens over 140 days (study endpoint) was observed only in the C3H\/HeN mouse strain when using pAAV\/1.2HBV-A and pMC\/1.0HBV-D with pre-C and pre-S recombination sites. In addition, pAAV\/1.2HBV-A in C3H\/HeN sustained HBV core antigen positivity up to the study endpoint in C3H\/HeN mice. Moreover, introducing the point mutation in the START codon of polymerase effectively prevented the formation of viral progeny. Our study establishes an accessible and affordable experimental paradigm for developing a robust mouse model reflecting CHB suitable for preclinical testing of anti-HBV therapeutics in a BSL2 animal facility.<\/jats:p>","DOI":"10.33549\/physiolres.934979","type":"journal-article","created":{"date-parts":[[2023,3,20]],"date-time":"2023-03-20T11:01:42Z","timestamp":1679310102000},"page":"37-47","source":"Crossref","is-referenced-by-count":0,"title":["Development and Characterization of a Chronic Hepatitis B Murine Model With a Mutation in the START Codon of an HBV Polymerase"],"prefix":"10.33549","volume":"72","author":[{"given":"L","family":"VANEKOVA","sequence":"first","affiliation":[{"name":"Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences, Prague, Czech Republic,"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"M","family":"POLIDAROVA","sequence":"additional","affiliation":[{"name":"Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences, Prague, Czech Republic,"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"V","family":"CHARVAT","sequence":"additional","affiliation":[{"name":"Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences, Prague, Czech Republic,"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Z","family":"VAVRINA","sequence":"additional","affiliation":[{"name":"Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences, Prague, Czech Republic,"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"V","family":"VEVERKA","sequence":"additional","affiliation":[{"name":"Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences, Prague, Czech Republic,"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"G","family":"BIRKUS","sequence":"additional","affiliation":[{"name":"Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences, Prague, Czech Republic,"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"A","family":"BRAZDOVA","sequence":"additional","affiliation":[{"name":"Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences, Prague, Czech Republic,"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"19169","published-online":{"date-parts":[[2023,2,28]]},"reference":[{"key":"ref0","doi-asserted-by":"publisher","unstructured":"1.\tSunbul M. Hepatitis B virus genotypes: Global distribution and clinical importance. World J Gastroenterol 2014;20:5427-5434. https:\/\/doi.org\/10.3748\/wjg.v20.i18.5427","DOI":"10.3748\/wjg.v20.i18.5427"},{"key":"ref1","unstructured":"2.\tWorld Health Organization. Global Hepatitis Report 2017;2017."},{"key":"ref2","doi-asserted-by":"publisher","unstructured":"3.\tKrajden M, McNabb G, Petric M. The laboratory diagnosis of hepatitis B virus. Can J Infect Dis Med Microbiol 2005;16:65-72. https:\/\/doi.org\/10.1155\/2005\/450574","DOI":"10.1155\/2005\/450574"},{"key":"ref3","doi-asserted-by":"publisher","unstructured":"4.\tHa HL, Shin HJ, Feitelson MA, Yu DY. Oxidative stress and antioxidants in hepatic pathogenesis. World J Gastroenterol 2010;16:6035-6043. https:\/\/doi.org\/10.3748\/wjg.v16.i48.6035","DOI":"10.3748\/wjg.v16.i48.6035"},{"key":"ref4","doi-asserted-by":"publisher","unstructured":"5.\tLiu L, Yang F. Application of modified mesenchymal stem cells transplantation in the treatment of liver injury. Physiol Res 2021;70:327-343. https:\/\/doi.org\/10.33549\/physiolres.934623","DOI":"10.33549\/physiolres.934623"},{"key":"ref5","doi-asserted-by":"publisher","unstructured":"6.\tFarghali H, Kgalalelo Kemelo M, Wojnarov\u00e1 L, Kutinov\u00e1 Canov\u00e1 N. In vitro and in vivo experimental hepatotoxic models in liver research: applications to the assessment of potential hepatoprotective drugs. Physiol Res 2016;65(Suppl 4):S417-S425. https:\/\/doi.org\/10.33549\/physiolres.933506","DOI":"10.33549\/physiolres.933506"},{"key":"ref6","doi-asserted-by":"publisher","unstructured":"7.\tAkbar SMF, Al Mahtab M, Aguilar JC, Yoshida O, Khan S, Penton E, Gerardo GN, Hiasa Y. The Safety and Efficacy of a Therapeutic Vaccine for Chronic Hepatitis B: A Follow-Up Study of Phase III Clinical Trial. Vaccines (Basel) 2022;10:45. https:\/\/doi.org\/10.3390\/vaccines10010045","DOI":"10.3390\/vaccines10010045"},{"key":"ref7","doi-asserted-by":"publisher","unstructured":"8.\tLeowattana W, Leowattana T. Chronic hepatitis B: New potential therapeutic drugs target. World J Virol 2022;11:57. https:\/\/doi.org\/10.5501\/wjv.v11.i1.57","DOI":"10.5501\/wjv.v11.i1.57"},{"key":"ref8","doi-asserted-by":"publisher","unstructured":"9.\tLiu T, Sun Q, Gu J, Cen S, Zhang Q. Characterization of the tenofovir resistance-associated mutations in the hepatitis B virus isolates across genotypes A to D. Antiviral Res 2022;203:105348. https:\/\/doi.org\/10.1016\/j.antiviral.2022.105348","DOI":"10.1016\/j.antiviral.2022.105348"},{"key":"ref9","doi-asserted-by":"publisher","unstructured":"10.\tWieland SF. The chimpanzee model for hepatitis B virus infection. Cold Spring Harb Perspect Med 2015;5:a021469. https:\/\/doi.org\/10.1101\/cshperspect.a021469","DOI":"10.1101\/cshperspect.a021469"},{"key":"ref10","doi-asserted-by":"publisher","unstructured":"11.\tSummers J, Smolec JM, Snyder R. A virus similar to human hepatitis B virus associated with hepatitis and hepatoma in woodchucks. Proc Natl Acad Sci U S A 1978;75:4533. https:\/\/doi.org\/10.1073\/pnas.75.9.4533","DOI":"10.1073\/pnas.75.9.4533"},{"key":"ref11","doi-asserted-by":"publisher","unstructured":"12.\tMason WS, Seal G, Summers J. Virus of Pekin ducks with structural and biological relatedness to human hepatitis B virus. J Virol 1980;36:829. https:\/\/doi.org\/10.1128\/jvi.36.3.829-836.1980","DOI":"10.1128\/jvi.36.3.829-836.1980"},{"key":"ref12","doi-asserted-by":"publisher","unstructured":"13.\tMarion PL, Oshiro LS, Regnery DC, Scullard GH, Robinson WS. A virus in Beechey ground squirrels that is related to hepatitis B virus of humans. Proc Natl Acad Sci U S A 1980;77:2941. https:\/\/doi.org\/10.1073\/pnas.77.5.2941","DOI":"10.1073\/pnas.77.5.2941"},{"key":"ref13","doi-asserted-by":"publisher","unstructured":"14.\tGuidotti LG, Matzke B, Schaller H, Chisari FV. High-level hepatitis B virus replication in transgenic mice. J Virol 1995;69:6158-6169. https:\/\/doi.org\/10.1128\/jvi.69.10.6158-6169.1995","DOI":"10.1128\/jvi.69.10.6158-6169.1995"},{"key":"ref14","doi-asserted-by":"publisher","unstructured":"15.\tGrompe M, Al-Dhalimy M, Finegold M, Ou CN, Burlingame T, Kennaway NG, Soriano P. Loss of fumarylacetoacetate hydrolase is responsible for the neonatal hepatic dysfunction phenotype of lethal albino mice. Genes Dev 1993;7:2298-2307. https:\/\/doi.org\/10.1101\/gad.7.12a.2298","DOI":"10.1101\/gad.7.12a.2298"},{"key":"ref15","doi-asserted-by":"crossref","unstructured":"16.\tWu LL, Wang HY, Chen PJ. Hydrodynamic HBV transfection mouse model. Methods Mol Biol 2017;1540:227-235. https:\/\/doi.org\/10.1007\/978-1-4939-6700-1_19","DOI":"10.1007\/978-1-4939-6700-1_19"},{"key":"ref16","doi-asserted-by":"publisher","unstructured":"17.\tYan Z, Zeng J, Yu Y, Xiang K, Hu H, Zhou X, Gu L, Wang L, Zhao J, Young JAT, Gao L. HBVcircle: A novel tool to investigate hepatitis B virus covalently closed circular DNA. J Hepatol 2017;66:1149-1157. https:\/\/doi.org\/10.1016\/j.jhep.2017.02.004","DOI":"10.1016\/j.jhep.2017.02.004"},{"key":"ref17","doi-asserted-by":"publisher","unstructured":"18.\tOrtega-Prieto AM, Cherry C, Gunn H, Dorner M. In vivo model systems for hepatitis B virus research. ACS Infect Dis 2019;5:688-702. https:\/\/doi.org\/10.1021\/acsinfecdis.8b00223","DOI":"10.1021\/acsinfecdis.8b00223"},{"key":"ref18","doi-asserted-by":"publisher","unstructured":"19.\tMilich DR, Leroux-Roels GG. Immunogenetics of the response to HBsAg vaccination. Autoimmun Rev 2003;2:248-257. https:\/\/doi.org\/10.1016\/S1568-9972(03)00031-4","DOI":"10.1016\/s1568-9972(03)00031-4"},{"key":"ref19","doi-asserted-by":"publisher","unstructured":"20.\tPeng XH, Ren XN, Chen LX, Shi BS, Xu CH, Fang Z, Liu X, Chen JL, Zhang XN, Hu YW, Zhou XH. High persistence rate of hepatitis B virus in a hydrodynamic injection-based transfection model in C3H\/HeN mice. World J Gastroenterol 2015;21:3527-3536. https:\/\/doi.org\/10.3748\/wjg.v21.i12.3527","DOI":"10.3748\/wjg.v21.i12.3527"},{"key":"ref20","doi-asserted-by":"publisher","unstructured":"21.\tSantantonio T, Jung MC, Pastore G, Angarano G, G\u00fcnther S, Will H. Familial clustering of HBV pre-C and pre-S mutants. J Hepatol 1997;26:221-227. https:\/\/doi.org\/10.1016\/S0168-8278(97)80034-7","DOI":"10.1016\/s0168-8278(97)80034-7"},{"key":"ref21","doi-asserted-by":"publisher","unstructured":"22.\tHuang LR, Wu HL, Chen PJ, Chen DS. An immunocompetent mouse model for the tolerance of human chronic hepatitis B virus infection. Proc Natl Acad Sci U S A 2006;103:17862-17867. https:\/\/doi.org\/10.1073\/pnas.0608578103","DOI":"10.1073\/pnas.0608578103"},{"key":"ref22","doi-asserted-by":"publisher","unstructured":"23.\tWang D, Tai PWL, Gao G. Adeno-associated virus vector as a platform for gene therapy delivery. Nat Rev Drug Discov 2019;18:358-378. https:\/\/doi.org\/10.1038\/s41573-019-0012-9","DOI":"10.1038\/s41573-019-0012-9"},{"key":"ref23","doi-asserted-by":"publisher","unstructured":"24.\tGuo X, Chen P, Hou X, Xu W, Wang D, Wang TY, Zhang L, Zheng G, Gao ZL, He CY, Zhou B, Chen ZY. The recombined cccDNA produced using minicircle technology mimicked HBV genome in structure and function closely. Sci Rep 2016;6:25552. https:\/\/doi.org\/10.1038\/srep25552","DOI":"10.1038\/srep25552"},{"key":"ref24","doi-asserted-by":"publisher","unstructured":"25.\tWang L, Cao M, Wei QL, Zhao ZH, Xiang Q, Wang HJ, Zhang HT, Lai GQ. A new model mimicking persistent HBV e antigen-negative infection using covalently closed circular DNA in immunocompetent mice. PLoS One 2017;12:e0175992. https:\/\/doi.org\/10.1371\/journal.pone.0175992","DOI":"10.1371\/journal.pone.0175992"},{"key":"ref25","doi-asserted-by":"publisher","unstructured":"26.\tChou HH, Chien WH, Wu LL, Cheng CH, Chung CH, Horng JH, Ni YH, ET AL. Age-related immune clearance of hepatitis B virus infection requires the establishment of gut microbiota. Proc Natl Acad Sci U S A 2015;112:2175-2180. https:\/\/doi.org\/10.1073\/pnas.1424775112","DOI":"10.1073\/pnas.1424775112"},{"key":"ref26","doi-asserted-by":"publisher","unstructured":"27.\tLi L, Li S, Zhou Y, Yang L, Zhou D, Yang Y, Lu M, Yang D, Song J. The dose of HBV genome contained plasmid has a great impact on HBV persistence in hydrodynamic injection mouse model. Virol J 2017;14:1-12. https:\/\/doi.org\/10.1186\/s12985-017-0874-6","DOI":"10.1186\/s12985-017-0874-6"},{"key":"ref27","doi-asserted-by":"crossref","unstructured":"28.\tSuda T, Liu D. Hydrodynamic delivery. Adv Genet 2015;89:89-111. https:\/\/doi.org\/10.1016\/bs.adgen.2014.10.002","DOI":"10.1016\/bs.adgen.2014.10.002"},{"key":"ref28","doi-asserted-by":"publisher","unstructured":"29.\tSun Y, Qi Y, Peng B, Li W. NTCP-reconstituted in vitro HBV infection system. Methods Mol Biol 2017;1540:1-14. https:\/\/doi.org\/10.1007\/978-1-4939-6700-1_1","DOI":"10.1007\/978-1-4939-6700-1_1"},{"key":"ref29","doi-asserted-by":"publisher","unstructured":"30.\tDu Y, Broering R, Li X, Zhang X, Liu J, Yang D, Lu M. In vivo mouse models for hepatitis B virus infection and their application. Front Immunol 2021;12:4578. https:\/\/doi.org\/10.3389\/fimmu.2021.766534","DOI":"10.3389\/fimmu.2021.766534"},{"key":"ref30","doi-asserted-by":"publisher","unstructured":"31.\t\u0158ez\u00e1\u010dov\u00e1 P, Pokorn\u00e1 J, Brynda J, Ko\u017e\u00ed\u0161ek M, C\u00edgler P, Lep\u0161\u00edk M, Fanfrl\u00edk J, ET AL. Design of HIV protease inhibitors based on inorganic polyhedral metallacarboranes. J Med Chem 2009;52:7132-7141. https:\/\/doi.org\/10.1021\/jm9011388","DOI":"10.1021\/jm9011388"},{"key":"ref31","doi-asserted-by":"publisher","unstructured":"32.\tKao JH. Diagnosis of hepatitis B virus infection through serological and virological markers. Expert Rev Gastroenterol Hepatol 2008;2:553-562. https:\/\/doi.org\/10.1586\/17474124.2.4.553","DOI":"10.1586\/17474124.2.4.553"},{"key":"ref32","doi-asserted-by":"publisher","unstructured":"33.\tKramvis A. Genotypes and genetic variability of hepatitis B virus. Intervirology 2014;57:141-150. https:\/\/doi.org\/10.1159\/000360947","DOI":"10.1159\/000360947"},{"key":"ref33","doi-asserted-by":"publisher","unstructured":"34.\tSengupta S, Rehman S, Durgapal H, Acharya SK, Panda SK. Role of surface promoter mutations in hepatitis B surface antigen production and secretion in occult hepatitis B virus infection. J Med Virol 2007;79:220-228. https:\/\/doi.org\/10.1002\/jmv.20790","DOI":"10.1002\/jmv.20790"},{"key":"ref34","doi-asserted-by":"publisher","unstructured":"35.\tZhou T, Guo H, Guo JT, Cuconati A, Mehta A, Block TM. Hepatitis B virus e antigen production is dependent upon covalently closed circular (ccc) DNA in HepAD38 cell cultures and may serve as a cccDNA surrogate in antiviral screening assays. Antiviral Res 2006;72:116-124. https:\/\/doi.org\/10.1016\/j.antiviral.2006.05.006","DOI":"10.1016\/j.antiviral.2006.05.006"},{"key":"ref35","doi-asserted-by":"publisher","unstructured":"36.\tLevrero M, Pollicino T, Petersen J, Belloni L, Raimondo G, Dandri M. Control of cccDNA function in hepatitis B virus infection. J Hepatol 2009;51:581-592. https:\/\/doi.org\/10.1016\/j.jhep.2009.05.022","DOI":"10.1016\/j.jhep.2009.05.022"},{"key":"ref36","unstructured":"37.\tThe World Health Organisation. Guidelines on Hepatitis B and C Testing, 2017."}],"container-title":["Physiological Research"],"original-title":[],"language":"en","deposited":{"date-parts":[[2023,3,20]],"date-time":"2023-03-20T11:02:09Z","timestamp":1679310129000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.biomed.cas.cz\/physiolres\/pdf\/2023\/72_37.pdf"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,2,28]]},"references-count":37,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2023,2,28]]}},"URL":"https:\/\/doi.org\/10.33549\/physiolres.934979","relation":{},"ISSN":["1802-9973","0862-8408"],"issn-type":[{"value":"1802-9973","type":"electronic"},{"value":"0862-8408","type":"print"}],"subject":[],"published":{"date-parts":[[2023,2,28]]}}}