Open AccessTherapeutic liver repopulation by transient acetaminophen selection of gene-modified hepatocytes
Author(s) -
Anne Vonada,
Amita Tiyaboonchai,
Sean Nygaard,
Jeffrey Posey,
A. Mack Peters,
Shelley R. Winn,
Alessio Cantore,
Luigi Naldini,
Cary O. Harding,
Markus Grompe
Publication year - 2021
Publication title -
science translational medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 6.819
H-Index - 216
eISSN - 1946-6242
pISSN - 1946-6234
DOI - 10.1126/scitranslmed.abg3047
Subject(s) - repopulation , acetaminophen , selection (genetic algorithm) , liver regeneration , gene , hepatocyte , genetic enhancement , pharmacology , transient (computer programming) , cancer research , biology , medicine , microbiology and biotechnology , chemistry , regeneration (biology) , genetics , computer science , stem cell , haematopoiesis , in vitro , artificial intelligence , operating system
Gene therapy by integrating vectors is promising for monogenic liver diseases, especially in children where episomal vectors remain transient. However, reaching the therapeutic threshold with genome-integrating vectors is challenging. Therefore, we developed a method to expand hepatocytes bearing therapeutic transgenes. The common fever medicine acetaminophen becomes hepatotoxic via cytochrome p450 metabolism. Lentiviral vectors with transgenes linked in cis to a Cypor shRNA were administered to neonatal mice. Hepatocytes lacking the essential cofactor of Cyp enzymes, NADPH-cytochrome p450 reductase (Cypor), were selected in vivo by acetaminophen administration, replacing up to 50% of the hepatic mass. Acetaminophen treatment of the mice resulted in over 30-fold expansion of transgene-bearing hepatocytes and achieved therapeutic thresholds in hemophilia B and phenylketonuria. We conclude that therapeutically modified hepatocytes can be selected safely and efficiently in preclinical models with a transient regimen of moderately hepatotoxic acetaminophen.