{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,12,31]],"date-time":"2025-12-31T04:36:57Z","timestamp":1767155817727,"version":"build-2238731810"},"reference-count":22,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2024,7,23]],"date-time":"2024-07-23T00:00:00Z","timestamp":1721692800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2024,7,23]],"date-time":"2024-07-23T00:00:00Z","timestamp":1721692800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["npj Viruses"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>With 254 million chronically infected patients, hepatitis B virus (HBV) continues to be a severe health threat. While animal models play a crucial role in developing new therapies, the availability of preclinical HBV models is very limited. Therefore, novel in vivo infection models are urgently needed. The bona fide HBV receptor, sodium-taurocholate cotransporting polypeptide (NTCP), determines HBV\u2019s species and cell-type specificity. Recent studies have indicated that the expression of human NTCP is the only limiting factor for HBV infection in selected species, such as macaques or pigs. Here, we confirm HBV infection of pig hepatocytes expressing human NTCP and show that porcine NTCP does not support HBV binding. By gradually humanizing porcine NTCP and site-directed mutagenesis, we identified amino acids 158 and 167 in porcine NTCP, limiting HBV interaction. In a proof-of-concept experiment, we showed that the expression of porcine NTCP with humanized amino acids 157-167 renders primary porcine hepatocytes fully susceptible to HBV. These results pave the way for generating transgenic pigs with humanized porcine chimeric NTCP as a novel, fully immunocompetent infection model for developing and validating new curative HBV therapies.<\/jats:p>","DOI":"10.1038\/s44298-024-00041-5","type":"journal-article","created":{"date-parts":[[2024,7,23]],"date-time":"2024-07-23T06:10:03Z","timestamp":1721715003000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Identification of amino acids restricting HBV receptor function in porcine NTCP"],"prefix":"10.1038","volume":"2","author":[{"given":"Samuel D.","family":"Jeske","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jochen M.","family":"Wettengel","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Florian","family":"Gegenfurtner","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Konrad","family":"Fischer","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Judith","family":"Moosm\u00fcller","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anindita","family":"Chakraborty","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chunkyu","family":"Ko","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Benjamin J.","family":"Burwitz","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Angelika","family":"Schnieke","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ulrike","family":"Protzer","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2024,7,23]]},"reference":[{"key":"41_CR1","unstructured":"World Health Organization. Global hepatitis report 2024: action for access in low- and middle-income countries. (World Health Organization, 2024). https:\/\/iris.who.int\/handle\/10665\/376461."},{"key":"41_CR2","doi-asserted-by":"publisher","first-page":"S41","DOI":"10.1016\/j.jhep.2016.02.009","volume":"64","author":"J Lucifora","year":"2016","unstructured":"Lucifora, J. & Protzer, U. Attacking hepatitis B virus cccDNA\u2013The holy grail to hepatitis B cure. J. Hepatol. 64, S41\u2013S48 (2016).","journal-title":"J. Hepatol."},{"key":"41_CR3","doi-asserted-by":"publisher","first-page":"325","DOI":"10.1053\/j.gastro.2018.10.032","volume":"156","author":"AJ Gehring","year":"2019","unstructured":"Gehring, A. J. & Protzer, U. Targeting innate and adaptive immune responses to cure chronic HBV infection. Gastroenterology 156, 325\u2013337 (2019).","journal-title":"Gastroenterology"},{"key":"41_CR4","doi-asserted-by":"publisher","first-page":"828","DOI":"10.3390\/v12080828","volume":"12","author":"JM Wettengel","year":"2020","unstructured":"Wettengel, J. M. & Burwitz, B. J. Innovative HBV animal models based on the entry receptor NTCP. Viruses 12, 828 (2020).","journal-title":"Viruses"},{"key":"41_CR5","doi-asserted-by":"publisher","unstructured":"Institute of Medicine and National Research Council. Chimpanzees in Biomedical and Behavioral Research: Assessing the Necessity. (The National Academies Press, Washington, DC, 2011). https:\/\/doi.org\/10.17226\/13257.","DOI":"10.17226\/13257"},{"key":"41_CR6","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.00049","volume":"1","author":"H Yan","year":"2012","unstructured":"Yan, H. et al. Sodium taurocholate cotransporting polypeptide is a functional receptor for human hepatitis B and D virus. eLife 1, e00049 (2012).","journal-title":"eLife"},{"key":"41_CR7","doi-asserted-by":"publisher","first-page":"175","DOI":"10.1038\/cmi.2013.66","volume":"11","author":"H Li","year":"2014","unstructured":"Li, H. et al. HBV life cycle is restricted in mouse hepatocytes expressing human NTCP. Cell. Mol. Immunol. 11, 175\u2013183 (2014).","journal-title":"Cell. Mol. Immunol."},{"key":"41_CR8","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1007\/978-1-4939-6700-1_1","volume":"1540","author":"Y Sun","year":"2017","unstructured":"Sun, Y., Qi, Y., Peng, B. & Li, W. NTCP-reconstituted in vitro HBV infection system. Methods Mol. Biol. 1540, 1\u201314 (2017).","journal-title":"Methods Mol. Biol."},{"key":"41_CR9","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-017-01953-y","volume":"8","author":"BJ Burwitz","year":"2017","unstructured":"Burwitz, B. J. et al. Hepatocytic expression of human sodium-taurocholate cotransporting polypeptide enables hepatitis B virus infection of macaques. Nat. Commun. 8, 2146 (2017).","journal-title":"Nat. Commun."},{"key":"41_CR10","doi-asserted-by":"publisher","first-page":"7977","DOI":"10.1128\/JVI.03540-12","volume":"87","author":"H Yan","year":"2013","unstructured":"Yan, H. et al. Molecular determinants of hepatitis B and D virus entry restriction in mouse sodium taurocholate cotransporting polypeptide. J. Virol. 87, 7977\u20137991 (2013).","journal-title":"J. Virol."},{"key":"41_CR11","doi-asserted-by":"publisher","DOI":"10.1016\/j.antiviral.2021.105140","volume":"194","author":"C Ko","year":"2021","unstructured":"Ko, C. et al. Intramolecular recombination enables the formation of hepatitis B virus (HBV) cccDNA in mice after HBV genome transfer using recombinant AAV vectors. Antivir. Res. 194, 105140 (2021).","journal-title":"Antivir. 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All other authors declare no financial or non-financial competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"30"}}