Adenovirus vector-transduced hepatocytes implanted via a preformed collagen/PTFE support persist for at least 4 weeks...

Liver-directed gene therapy has the potential to provide an effective adjutant to conventional treatments for liver diseases. In vivo gene transfer is promising but still effectively out of reach with conventional gene delivery techniques. Ex vivo gene therapy using hepatocyte transplantation, however, has been encouraging. We describe and approach toward treatment of liver-related diseases by combining several of the advantageous properties of current liver-directed therapies. Primary hepatocytes were isolated, transduced with an adenovirus vector encoding a reporter gene, embedded in a collagen/polytetrafluoroethylene (PTFE) lattice, and implanted in mice. Recovered 'hepatocyte-organoids' were assayed for the presence and viability of the implanted hepatocytes, duration of transgene expression and presence of the adenovirus vector. In an initial attempt, we demonstrate that genetically modified hepatocytes can survive and express a transgene for at least 4 weeks in vivo when embedded in a collagen/PTFE support and implanted in the intraperitoneal cavity. This approach takes advantage of hepatocyte-specific functions in order to treat diseases where a fraction of the normal enzymatic activity is sufficient to alleviate a disease phenotype.