{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T20:12:04Z","timestamp":1768507924912,"version":"3.49.0"},"posted":{"date-parts":[[2023,8,28]]},"group-title":"Genetics","reference-count":38,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2023,8,28]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>Hepatocyte transplantation for genetic liver diseases has several potential advantages over gene therapy. However, low efficiency of cell engraftment has limited its clinical implementation. This problem could be overcome by selectively expanding transplanted donor cells until they replace enough of the liver mass to achieve therapeutic benefit. We previously described a gene therapy method to selectively expand hepatocytes deficient in cytochrome p450 reductase (Cypor) using acetaminophen (APAP). Because Cypor is required for the transformation of APAP to a hepatotoxic metabolite, Cypor deficient cells are protected from toxicity and are able to expand following APAP-induced liver injury. Here, we apply this selection system to correct a mouse model of phenylketonuria (PKU) by cell transplantation. Hepatocytes from a wildtype donor animal were edited in vitro to create Cypor deficiency and then transplanted into PKU animals. Following selection with APAP, blood phenylalanine concentrations were fully normalized and remained stable following APAP withdrawal. Cypor-deficient hepatocytes expanded from &lt;1% to \u223c14% in corrected animals, and they showed no abnormalities in blood chemistries, liver histology, or drug metabolism. We conclude that APAP-mediated selection of transplanted hepatocytes is a potential therapeutic for PKU with long-term efficacy and a favorable safety profile.<\/jats:p>","DOI":"10.1101\/2023.08.27.554228","type":"posted-content","created":{"date-parts":[[2023,8,28]],"date-time":"2023-08-28T21:45:12Z","timestamp":1693259112000},"source":"Crossref","is-referenced-by-count":1,"title":["Complete correction of murine phenylketonuria by selection-enhanced hepatocyte transplantation"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0398-1898","authenticated-orcid":false,"given":"Anne","family":"Vonada","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5623-4285","authenticated-orcid":false,"given":"Leslie","family":"Wakefield","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6577-9420","authenticated-orcid":false,"given":"Michael","family":"Martinez","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9146-3112","authenticated-orcid":false,"given":"Cary O.","family":"Harding","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6616-4345","authenticated-orcid":false,"given":"Markus","family":"Grompe","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0116-7393","authenticated-orcid":false,"given":"Amita","family":"Tiyaboonchai","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2023090103250529000_2023.08.27.554228v1.1","doi-asserted-by":"publisher","DOI":"10.1038\/pr.2017.284"},{"key":"2023090103250529000_2023.08.27.554228v1.2","doi-asserted-by":"publisher","DOI":"10.1007\/s10545-006-0245-8"},{"key":"2023090103250529000_2023.08.27.554228v1.3","first-page":"14.12.11","article-title":"The history and use of human hepatocytes for the treatment of liver diseases: the first 100 patients","volume":"62","year":"2014","journal-title":"Curr Protoc Toxicol"},{"key":"2023090103250529000_2023.08.27.554228v1.4","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(02)07529-3"},{"key":"2023090103250529000_2023.08.27.554228v1.5","doi-asserted-by":"publisher","DOI":"10.1053\/j.gastro.2006.01.008"},{"key":"2023090103250529000_2023.08.27.554228v1.6","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM199805143382004"},{"key":"2023090103250529000_2023.08.27.554228v1.7","doi-asserted-by":"crossref","unstructured":"Vonada A , Tiyaboonchai A , Nygaard S , Posey J , Peters AM , Winn SR , Cantore A , et al. Therapeutic liver repopulation by transient acetaminophen selection of gene-modified hepatocytes. Sci Transl Med 2021;13.","DOI":"10.1126\/scitranslmed.abg3047"},{"key":"2023090103250529000_2023.08.27.554228v1.8","doi-asserted-by":"publisher","DOI":"10.14218\/JCTH.2015.00052"},{"key":"2023090103250529000_2023.08.27.554228v1.9","doi-asserted-by":"publisher","DOI":"10.1016\/j.abb.2012.09.002"},{"key":"2023090103250529000_2023.08.27.554228v1.10","doi-asserted-by":"crossref","first-page":"7391","DOI":"10.1038\/s41467-022-35097-5","article-title":"Self-cleaving guide RNAs enable pharmacological selection of precise gene editing events in vivo","volume":"13","year":"2022","journal-title":"Nat Commun"},{"key":"2023090103250529000_2023.08.27.554228v1.11","doi-asserted-by":"publisher","DOI":"10.1038\/gim.2013.157"},{"key":"2023090103250529000_2023.08.27.554228v1.12","doi-asserted-by":"publisher","DOI":"10.3727\/096368912X653255"},{"key":"2023090103250529000_2023.08.27.554228v1.13","doi-asserted-by":"crossref","first-page":"987","DOI":"10.1016\/j.jhep.2016.12.017","article-title":"Host conditioning and rejection monitoring in hepatocyte transplantation in humans","volume":"66","year":"2017","journal-title":"J Hepatol"},{"key":"2023090103250529000_2023.08.27.554228v1.14","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1016\/j.ymthe.2005.03.025","article-title":"Low therapeutic threshold for hepatocyte replacement in murine phenylketonuria","volume":"12","year":"2005","journal-title":"Mol Ther"},{"key":"2023090103250529000_2023.08.27.554228v1.15","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1016\/j.ymgme.2011.07.027","article-title":"Hepatocytes from wild-type or heterozygous donors are equally effective in achieving successful therapeutic liver repopulation in murine phenylketonuria (PKU)","volume":"104","year":"2011","journal-title":"Mol Genet Metab"},{"key":"2023090103250529000_2023.08.27.554228v1.16","doi-asserted-by":"publisher","DOI":"10.1002\/dvg.20335"},{"key":"2023090103250529000_2023.08.27.554228v1.17","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gku936"},{"key":"2023090103250529000_2023.08.27.554228v1.18","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.88.4.1217"},{"key":"2023090103250529000_2023.08.27.554228v1.19","doi-asserted-by":"crossref","first-page":"1205","DOI":"10.1093\/genetics\/134.4.1205","article-title":"Mouse models of human phenylketonuria","volume":"134","year":"1993","journal-title":"Genetics"},{"key":"2023090103250529000_2023.08.27.554228v1.20","doi-asserted-by":"crossref","first-page":"306","DOI":"10.1016\/j.ymgme.2020.09.005","article-title":"A novel Pah-exon1 deleted murine model of phenylalanine hydroxylase (PAH) deficiency","volume":"131","year":"2020","journal-title":"Mol Genet Metab"},{"key":"2023090103250529000_2023.08.27.554228v1.21","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M212087200"},{"key":"2023090103250529000_2023.08.27.554228v1.22","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M303125200"},{"key":"2023090103250529000_2023.08.27.554228v1.23","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1055\/s-0041-1742279","article-title":"The Role of Liver Zonation in Physiology, Regeneration, and Disease","volume":"42","year":"2022","journal-title":"Semin Liver Dis"},{"key":"2023090103250529000_2023.08.27.554228v1.24","doi-asserted-by":"publisher","DOI":"10.1097\/00007691-200008000-00008"},{"key":"2023090103250529000_2023.08.27.554228v1.25","doi-asserted-by":"publisher","DOI":"10.1006\/taap.1998.8543"},{"key":"2023090103250529000_2023.08.27.554228v1.26","doi-asserted-by":"publisher","DOI":"10.1038\/clpt.1993.63"},{"key":"2023090103250529000_2023.08.27.554228v1.27","doi-asserted-by":"publisher","DOI":"10.1124\/dmd.112.048991"},{"key":"2023090103250529000_2023.08.27.554228v1.28","doi-asserted-by":"publisher","DOI":"10.1124\/dmd.119.087718"},{"key":"2023090103250529000_2023.08.27.554228v1.29","doi-asserted-by":"crossref","unstructured":"Hamilton BA , Wright JF . Challenges Posed by Immune Responses to AAV Vectors: Addressing Root Causes. Front Immunol 2021;12:675897.","DOI":"10.3389\/fimmu.2021.675897"},{"key":"2023090103250529000_2023.08.27.554228v1.30","doi-asserted-by":"publisher","DOI":"10.1159\/000369552"},{"key":"2023090103250529000_2023.08.27.554228v1.31","doi-asserted-by":"publisher","DOI":"10.1038\/mt.2008.72"},{"key":"2023090103250529000_2023.08.27.554228v1.32","doi-asserted-by":"publisher","DOI":"10.1038\/mt.2011.151"},{"key":"2023090103250529000_2023.08.27.554228v1.33","doi-asserted-by":"publisher","DOI":"10.1006\/mgme.2001.3198"},{"key":"2023090103250529000_2023.08.27.554228v1.34","doi-asserted-by":"publisher","DOI":"10.1159\/000114857"},{"key":"2023090103250529000_2023.08.27.554228v1.35","doi-asserted-by":"publisher","DOI":"10.1530\/EJE-10-0764"},{"key":"2023090103250529000_2023.08.27.554228v1.36","doi-asserted-by":"publisher","DOI":"10.1080\/03602530902843483"},{"key":"2023090103250529000_2023.08.27.554228v1.37","doi-asserted-by":"crossref","first-page":"374","DOI":"10.1016\/j.apsb.2016.07.012","article-title":"An update on the role of intestinal cytochrome P450 enzymes in drug disposition","volume":"6","year":"2016","journal-title":"Acta Pharm Sin B"},{"key":"2023090103250529000_2023.08.27.554228v1.38","doi-asserted-by":"crossref","first-page":"234","DOI":"10.1016\/j.omtm.2019.12.004","article-title":"AAV-Mediated CRISPR\/Cas9 Gene Editing in Murine Phenylketonuria","volume":"17","year":"2020","journal-title":"Mol Ther Methods Clin Dev"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/2023.08.27.554228","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T05:08:22Z","timestamp":1768453702000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.1101\/2023.08.27.554228"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,8,28]]},"references-count":38,"URL":"https:\/\/doi.org\/10.1101\/2023.08.27.554228","relation":{"is-preprint-of":[{"id-type":"doi","id":"10.1097\/HEP.0000000000000631","asserted-by":"subject"}]},"subject":[],"published":{"date-parts":[[2023,8,28]]},"subtype":"preprint"}}