Gene therapy successfully delays degeneration in a mouse model of PDE6A-linked retinitis pigmentosa (RP 43).

Retinitis pigmentosa type 43 (RP43) is a blinding disease caused by mutations in the gene for rod phosphodiesterase 6 alpha (PDE6A). The disease process begins with a dysfunction of rod photoreceptors, subsequently followed by a currently untreatable progressive degeneration of the entire outer retina. Aiming at a curative approach via PDE6A gene supplementation, we developed a novel adeno-associated viral (AAV) vector for expression of the human PDE6A cDNA under control of the human rhodopsin promotor (rAAV8.PDE6A). In this study, we assessed the therapeutic efficacy of rAAV8.PDE6A in the Pde6anmf363/nmf363 mutant mouse model of RP43. All mice included in this study were treated with subretinal injections of the vector at 2 weeks after birth. The therapeutic effect was monitored at one month and at six months post injection. Biological function of the transgene was assessed in vivo by means of electroretinography (ERG). The degree of morphological rescue was investigated both in vivo using optical coherence tomography (OCT) and ex vivo by immunohistological stainings. We found that the novel rAAV8.PDE6A vector resulted in a stable and efficient expression of PDE6A protein in rod photoreceptors of Pde6anmf363/nmf363 mice following treatment at both the short and long-term time-points. The treatment led to a substantial morphological preservation of outer nuclear layer thickness, rod outer segment structure, and prolonged survival of cone photoreceptors for at least up to 6 months. Additionally, the ERG analysis confirmed a restoration of retinal function in a group of treated mice. Taken together, we provide successful proof-of-concept for the cross-species efficacy of the rAAV8.PDE6A vector developed for use in human patients. Importantly, our data show stable expression and rescue effects for a prolonged period of time, raising hope for future translational studies based on this approach.