Novel strategies to improve viral gene targeting and therapeutic liver repopulation in vivo

Orthotopic liver transplantation effectively treats pediatric metabolic diseases caused by hepatocyte-specific enzyme deficiencies, but donor scarcity, high cost, and lifelong immunosuppression limit its use. This dissertation explores alternative strategies: gene therapy and hepatocyte transplantation. First, recombinant adeno-associated viral (rAAV) vectors achieved stable in vivo gene targeting in neonatal and adult mice, with enhanced efficiency via transient inhibition of non-homologous end joining using vanillin and proteasome inhibition with bortezomib. Second, selective hepatocyte repopulation was demonstrated using CEHPOBA, enabling transient positive selection in wild-type livers. These findings highlight promising approaches for correcting metabolic liver disorders without whole-organ transplantation.