title: A comparison of AAV strategies distinguishes overlapping vectors for efficient systemic delivery of the 6.2 kb Dysferlin coding sequence.
abstract: Recombinant adeno-associated virus (rAAV) is currently the best vector for gene delivery into the skeletal muscle. However, the 5-kb packaging size of this virus is a major obstacle for large gene transfer. This past decade, many different strategies were developed to circumvent this issue (concatemerization-splicing, overlapping vectors, hybrid dual or fragmented AAV). Loss of function mutations in the DYSF gene whose coding sequence is 6.2kb lead to progressive muscular dystrophies (LGMD2B: OMIM_253601; MM: OMIM_254130; DMAT: OMIM_606768). In this study, we compared large gene transfer techniques to deliver the DYSF gene into the skeletal muscle. After rAAV8s intramuscular injection into dysferlin deficient mice, we showed that the overlap strategy is the most effective approach to reconstitute a full-length messenger. After systemic administration, the level of dysferlin obtained on different muscles corresponded to 0.5- to 2-fold compared to the normal level. We further demonstrated that the overlapping vector set was efficient to correct the histopathology, resistance to eccentric contractions and whole body force in the dysferlin deficient mice. Altogether, these data indicate that using overlapping vectors could be a promising approach for a potential clinical treatment of dysferlinopathies.
authors.affiliation: Inserm, U951 , Evry, France ; Genethon, R&D Department, INTEGRARE Research Unit , Evry, France.
authors.forename: Marina
authors.initials: M
authors.lastname: Pryadkina
authors.affiliation: Inserm, U951 , Evry, France ; Genethon, R&D Department, INTEGRARE Research Unit , Evry, France.
authors.forename: William
authors.initials: W
authors.lastname: Lostal
authors.affiliation: Inserm, U951 , Evry, France ; Genethon, R&D Department, INTEGRARE Research Unit , Evry, France.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Bourg
authors.affiliation: Inserm, U951 , Evry, France ; Genethon, R&D Department, INTEGRARE Research Unit , Evry, France.
authors.forename: Karine
authors.initials: K
authors.lastname: Charton
authors.affiliation: Inserm, U951 , Evry, France ; Genethon, R&D Department, INTEGRARE Research Unit , Evry, France.
authors.forename: Carinne
authors.initials: C
authors.lastname: Roudaut
authors.affiliation: Gene Therapy Center, University of North Carolina , Chapel Hill, North Carolina, USA ; Department of Ophthalmology, University of North Carolina , Chapel Hill, North Carolina, USA.
authors.forename: Matthew L
authors.initials: ML
authors.lastname: Hirsch
authors.affiliation: Inserm, U951 , Evry, France ; Genethon, R&D Department, INTEGRARE Research Unit , Evry, France.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Richard