Adeno-associated virus vector gene transfer and sarcolemmal expression of a 144 kDa micro-dystrophin effectively restores the dystrophin-associated protein complex and inhibits myofibre degeneration in nude/mdx mice

被引:70
作者
Fabb, SA
Wells, DJ
Serpente, P
Dickson, G [1 ]
机构
[1] Univ London, Royal Holloway & Bedford New Coll, Sch Biol Sci, Ctr Biomed Sci, Egham TW20 0EX, Surrey, England
[2] Charing Cross Hosp, Imperial Coll Fac Med, Dept Neuromuscular Dis, Div Neurosci & Psychol Med,Gene Targeting Unit, London W6 8RP, England
关键词
D O I
10.1093/hmg/11.7.733
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Duchenne muscular dystrophy is a severe life-threatening X-linked recessive disorder, caused by mutations in the dystrophin gene, for which currently there is no effective treatment. Because of the large size of the dystrophin cDNA (14 kb) this precluded it from being used in early adenovirus- or retrovirus-based gene therapy vectors. However, some therapeutic success has been achieved in mdx mice using adenovirus- and retrovirus-mediated transfer of a 6.3 kb recombinant mini-dystrophin cDNA. Despite this, problems with immunogenicity and inefficient transduction of mature myofibres make these vectors less than ideal for gene transfer to skeletal muscle. Adeno-associated viral (AAV) vectors overcome many of the problems associated with other vector systems. However, AAV vectors can only accommodate <5 kb of foreign DNA. For this reason we have produced a micro-dystrophin cDNA gene construct that is <3.8 kb. This construct, driven by a CMV promoter, was introduced into the skeletal muscle of 12-day-old nude/mdx mice using an AAV vector, resulting in specific sarcolemmal expression of micro-dystrophin in >50% of myofibres up to 20 weeks of age, and effective restoration of the dystrophin-associated protein (DAP) complex components. Additionally, evaluation of central nucleation indicated a significant inhibition of degenerative dystrophic muscle pathology. We have therefore shown that the current micro-dystrophin gene delivered in vivo using an AAV vector is not only capable of restoring sarcolemmal DAP complexes, but can also ameliorate dystrophic pathology at the cellular level.
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页码:733 / 741
页数:9
相关论文
共 68 条
  • [1] HUMAN DYSTROPHIN EXPRESSION IN MDX MICE AFTER INTRAMUSCULAR INJECTION OF DNA CONSTRUCTS
    ACSADI, G
    DICKSON, G
    LOVE, DR
    JANI, A
    WALSH, FS
    GURUSINGHE, A
    WOLFF, JA
    DAVIES, KE
    [J]. NATURE, 1991, 352 (6338) : 815 - 818
  • [2] Dystrophin expression in muscles of mdx mice after adenovirus-mediated in vivo gene transfer
    Acsadi, G
    Lochmuller, H
    Jani, A
    Huard, J
    Massie, B
    Prescott, S
    Simoneau, M
    Petrof, BJ
    Karpati, G
    [J]. HUMAN GENE THERAPY, 1996, 7 (02) : 129 - 140
  • [3] Flexible hinges in dystrophin.
    Ahmed, N
    Man, NT
    Morris, GE
    [J]. BIOCHEMICAL SOCIETY TRANSACTIONS, 1998, 26 (03) : S310 - S310
  • [4] Akkaraju GR, 1999, J GENE MED, V1, P280, DOI 10.1002/(SICI)1521-2254(199907/08)1:4<280::AID-JGM45>3.0.CO
  • [5] 2-L
  • [6] A cluster of basic repeats in the dystrophin rod domain binds F-actin through an electrostatic interaction
    Amann, KJ
    Renley, BA
    Ervasti, JM
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 1998, 273 (43) : 28419 - 28423
  • [7] CLINICAL-MOLECULAR CORRELATION IN 104 MILD X-LINKED MUSCULAR-DYSTROPHY PATIENTS - CHARACTERIZATION OF SUBCLINICAL PHENOTYPES
    ANGELINI, C
    FANIN, M
    PEGORARO, E
    FREDA, MP
    CADALDINI, M
    MARTINELLO, F
    [J]. NEUROMUSCULAR DISORDERS, 1994, 4 (04) : 349 - 358
  • [8] In vivo targeted repair of a point mutation in the canine dystrophin gene by a chimeric RNA/DNA oligonucleotide
    Bartlett, RJ
    Stockinger, S
    Denis, MM
    Bartlett, WT
    Inverardi, L
    Le, TT
    Man, NT
    Morris, GE
    Bogan, DJ
    Metcalf-Bogan, J
    Kornegay, JN
    [J]. NATURE BIOTECHNOLOGY, 2000, 18 (06) : 615 - 622
  • [9] BEGGS AH, 1991, AM J HUM GENET, V49, P54
  • [10] Chamberlain JS, 1997, SOC GEN PHY, V52, P19