Gene Therapy with Mini-Dysferlin
The dysferlin protein domains that mediate function are not yet defined and the associated pathway is not well understood. New methods for intravascular and systemic delivery of some viral vectors (e.g. AAV6, AAV8) make gene therapy an attractive option. To overcome the limited size carrying capacity of these promising vectors, it is necessary to generate a partial but functional form of the Dysferlin gene. The three steps are therefore - creating partial proteins, testing to see which one is best and then using that to treat a mouse model for the disease. The approach uses partial dysferlin protein for studies to see if it moves locates and binds to the same areas within the cell as the full protein. In vitro studies will determine if this partial protein retains the same biochemical properties as the full protein and functions like it to repair cell membrane. Finally, the studies will test its ability, when packaged in the muscle-tropic vectors AAV6 and AAV8, to restore membrane repair in vivo in an established Miyoshi mouse model. Finally, the studies will determine the feasibility of and prove the efficacy of gene delivery in mouse models. If successful, the experiments will be repeated in other higher vertebrates before clinical trials in humans.