AAV-mediated gene transfer for hemophilia.

The goal of our work has been to establish an experimental basis for gene transfer as a method of treating hemophilia, an inherited bleeding disorder that results from the absence of functional factor VIII or factor IX. Using an adeno-associated viral vector derived from AAV serotype 2, we have shown in mice and in hemophilic dogs that we can achieve long-term expression (>3 years) of clotting factor at levels that would result in an improvement of clinical symptoms of the disease. A phase I trial of intramuscular injection of AAV-F.IX showed no evidence of local or systemic toxicity in any of the subjects. Muscle biopsies showed evidence for gene transfer and expression by polymerase chain reaction, Southern blot, and immunohistochemistry. We have also shown that AAV-F.IX can be delivered into the portal veins of hemophilic dogs and that this results in high circulating levels of factor IX, on the order of 5% to 14%, whereas delivery of similar doses to skeletal muscle results in factor levels of only 1% to 2%. Based on these results, a trial of AAV-mediated liver-directed gene transfer for hemophilia B has been proposed and is reviewed here.