Chimeric adeno-associated virus/antisense U1 small nuclear RNA effectively rescues dystrophin synthesis and muscle function by local treatment of mdx mice.
Duchenne muscular dystrophy (DMD) is a X-linked myopathy in which deletions and point mutations in the dystrophin gene abolish dystrophin expression. The defect can often be corrected at the posttranscriptional level by exon skipping. In an animal model of DMD, the mdx mouse, a point mutation in exon 23 of the dystrophin gene introduces a premature stop codon. Skipping of this exon reestablishes the open reading frame in the dystrophin mRNA. We have obtained persistent exon skipping in mdx mice by local muscle injection of AAV vectors expressing antisense sequences fused to either U1 or U7 small nuclear RNA (snRNA). In the transduced muscles, dystrophin expression, amelioration of muscle morphology, and significant force recovery were obtained. These data indicate that the expression of antisense snRNAs, combined with their efficient muscular delivery through AAV vectors, is a powerful strategy for the therapeutic treatment of DMD. Like U7 snRNA, spliceosomal U1 snRNA is also a suitable backbone for the expression of antisense molecules active in exon skipping.
Denti
Michela Alessandra
MA
Department of Genetics and Molecular Biology, Institute Pasteur Cenci-Bolognetti, and IBPM, University La Sapienza, 00185 Rome, Italy.
Rosa
Alessandro
A
D'Antona
Giuseppe
G
Sthandier
Olga
O
De Angelis
Fernanda Gabriella
FG
Nicoletti
Carmine
C
Allocca
Mariacarmela
M
Pansarasa
Orietta
O
Parente
Valeria
V
Musarò
Antonio
A
Auricchio
Alberto
A
Bottinelli
Roberto
R
Bozzoni
Irene
I
D000818:Animals; D001483:Base Sequence; D004274:DNA, Recombinant; D000229:Dependovirus / Q000235:genetics*; D016189:Dystrophin / Q000378:metabolism*; D015316:Genetic Therapy; D005822:Genetic Vectors; D007273:Injections, Intramuscular; D051379:Mice; D018101:Mice, Inbred mdx / Q000235:genetics; D008969:Molecular Sequence Data; D018485:Muscle Fibers, Skeletal / Q000502:physiology; D009137:Muscular Dystrophy, Animal / Q000628:therapy*; D016372:RNA, Antisense / Q000627:therapeutic use; D012342:RNA, Small Nuclear / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000627:therapeutic use
D004274:DNA, Recombinant; D016189:Dystrophin; D016372:RNA, Antisense; D012342:RNA, Small Nuclear; D011993:Recombinant Fusion Proteins; C405483:U1 small nuclear RNA; C405487:U7 small nuclear RNA