Efficient recovery of dysferlin deficiency by dual adeno-associated vector-mediated gene transfer.
Deficiency of the dysferlin protein presents as two major clinical phenotypes: limb-girdle muscular dystrophy type 2B and Miyoshi myopathy. Dysferlin is known to participate in membrane repair, providing a potential hypothesis to the underlying pathophysiology of these diseases. The size of the dysferlin cDNA prevents its direct incorporation into an adeno-associated virus (AAV) vector for therapeutic gene transfer into muscle. To bypass this limitation, we split the dysferlin cDNA at the exon 28/29 junction and cloned it into two independent AAV vectors carrying the appropriate splicing sequences. Intramuscular injection of the corresponding vectors into a dysferlin-deficient mouse model led to the expression of full-length dysferlin for at least 1 year. Importantly, systemic injection in the tail vein of the two vectors led to a widespread although weak expression of the full-length protein. Injections were associated with an improvement of the histological aspect of the muscle, a reduction in the number of necrotic fibers, restoration of membrane repair capacity and a global improvement in locomotor activity. Altogether, these data support the use of such a strategy for the treatment of dysferlin deficiency.
Lostal
William
W
Généthon, CNRS UMR8587 LAMBE, 1, rue de l'Internationale, 91000 Evry, France.
Bartoli
Marc
M
Bourg
Nathalie
N
Roudaut
Carinne
C
Bentaïb
Azeddine
A
Miyake
Katsuya
K
Guerchet
Nicolas
N
Fougerousse
Françoise
F
McNeil
Paul
P
Richard
Isabelle
I
D000818:Animals; D003433:Crosses, Genetic; D000229:Dependovirus / Q000235:genetics*; D000073939:Dysferlin; D005260:Female; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics*; D007273:Injections, Intramuscular; D008297:Male; D008565:Membrane Proteins / Q000172:deficiency* / Q000235:genetics / Q000627:therapeutic use*; D008566:Membranes / Q000473:pathology; D051379:Mice; D008810:Mice, Inbred C57BL; D009124:Muscle Proteins / Q000172:deficiency* / Q000235:genetics / Q000627:therapeutic use*; D018482:Muscle, Skeletal / Q000473:pathology; D049288:Muscular Dystrophies, Limb-Girdle / Q000235:genetics* / Q000628:therapy; D009154:Mutation; D010641:Phenotype; D019076:Transgenes; D014945:Wound Healing
C114839:DYSF protein, human; D000073939:Dysferlin; D008565:Membrane Proteins; D009124:Muscle Proteins