Original article| Volume 27, ISSUE 6, P400-405, September 2005

A novel approach to identify Duchenne muscular dystrophy patients for aminoglycoside antibiotics therapy


      Aminoglycoside antibiotics have been found to suppress nonsense mutations located in the defective dystophin gene in mdx mice, suggesting a possible treatment for Duchenne muscular dystrophy (DMD). However, it is very difficult to find patients that are applicable for this therapy, because: (1) only 5‐13% of DMD patients have nonsense mutations in the dystrophin gene, (2) it is challenging to find nonsense mutations in the gene because dystrophin cDNA is very long (14 kb), and (3) the efficiency of aminoglycoside-induced read-through is dependent on the kind of nonsense mutation. In order to develop a system for identifying candidates that qualify for aminoglycoside therapy, fibroblasts from nine DMD patients with nonsense mutation of dystrophin gene were isolated, induced to differentiate to myogenic lineage by AdMyoD, and exposed with gentamicin. The dystrophin expression in gentamicin-exposed myotubes was monitored by in vitro dystrophin staining and western blotting analysis. The results showed that gentamicin was able to induce dystrophin expression in the differentiated myotubes by the read-through of the nonsense mutation TGA in the gene; a read-through of the nonsense mutations TAA and TAG did not occur and consequently did not lead to dystrophin expression. Therefore, it is speculated that the aminoglycoside treatment is far more effective for DMD patients that have nonsense mutation TGA than for patients that have nonsense mutation TAA and TAG. In this study, we introduce an easy system to identify patients for this therapy and report for the first time, that dystrophin expression was detected in myotubes of DMD patients using gentamicin.


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        • Hoffman E.P.
        • Brown R.H.J.
        • Kunkel L.M.
        Dystrophin: the protein product of the Duchenne muscular dystrophy locus.
        Cell. 1987; 51: 919-928
        • Koenig M.
        • Hoffman E.P.
        • Bertelson C.J.
        • Monaca A.P.
        • Feener C.
        • Kunkel L.M.
        Complete cloning of the Duchenne muscular dystrophy (DMD) cDNA and preliminary genomic organization of the DMD gene in normal and affected individuals.
        Cell. 1987; 50: 509-517
        • Gillard E.F.
        • Chamberlain J.S.
        • Murphy E.G.
        • Duff C.L.
        • Smith B.
        • Burghes A.H.
        • et al.
        Molecular and phenotypic analysis of patients with deletions within the deletion-rich region of Duchenne muscular dystrophy (DMD) gene.
        Am J Hum Genet. 1989; 45: 368-372
        • Forrest S.M.
        • Cross G.S.
        • Flint T.
        • Speer A.
        • Robson K.J.
        • Davies K.E.
        Further studies of gene deletions that cause Duchenne and Becker muscular dystrophies.
        Genomics. 1988; 2: 109-114
        • Lindlof M.
        • Kiuru A.
        • Kaariainen H.
        • Kalimo H.
        • Lang H.
        • Pihko H.
        • et al.
        Gene deletion in X-linked muscular dystrophy.
        Am J Hum Genet. 1989; 44: 397-401
        • Prior T.
        • Bartolo C.
        • Pearl D.
        Spectrum of small mutations in the dystrophin coding region.
        Am J Hum Genet. 1995; 57: 22-33
        • Mendell J.R.
        • Buzin C.H.
        • Feng J.
        • Yan J.
        • Serrano C.
        • Sangani D.S.
        • et al.
        Diagnosis of Duchenne dystrophy by enhanced detection of small mutations.
        Neurology. 2001; 28: 645-650
        • Barton-Davis E.R.
        • Cordier L.
        • Shoturma D.I.
        • Leland S.E.
        • Sweeney H.L.
        Aminoglycoside antibiotics restore dystrophin function to skeletal muscle of mdx mice.
        J Clin Invest. 1999; 104: 375-381
        • Wagner R.W.
        • Hamed S.
        • Hadley D.W.
        • et al.
        Gentamicin treatment of Duchenne and Becker muscular dystrophy due to nonsense mutations.
        Ann Neurol. 2001; 49: 706-711
        • Howard M.T.
        • Shirts B.H.
        • Petros L.M.
        • Flanigan K.M.
        • Gesteland R.F.
        • Atkins J.F.
        Sequence specificity of aminoglycoside-induced stop codon read-through: potential implications for treatment of Duchenne muscular dystrophy.
        Ann Neurol. 2000; 48: 164-169
        • Pinney D.F.
        • SH P-W.
        • Konieczny S.F.
        Myogenic lineage determination and differentiation: evidence for a regulatory gene pathway.
        Cell. 1988; 53: 93
        • Niwa H.
        • Yamamura K.
        • Miyazaki J.
        Efficient selection for high-expression transfectants with a novel eukaryotic vector.
        Gene. 1991; 108: 1933-2000
        • Kimura S.
        • Fujishita S.
        • Ikezawa M.
        • Ogawa M.
        • Abe Kand Miike T.
        Muscle type promoter and its first intron abnormalities in dystrophin gene in patients with Duchenne muscular dystrophy.
        J Child Neur. 1998; 13: 202-290
        • Kimura S.
        • Ikezawa M.
        • Pruchnic R.
        • Balkir L.
        • Qu Z.
        • Lowenstein J.
        • et al.
        Persistent gene transfer to skeletal muscle mediated by stable transfected early myogenic progenitor cell.
        Basic Appl Myo. 2000; 10: 237-248
        • Andrew H.A.
        • Kunkel L.M.
        The structural and functional diversity of dystrophin.
        Nat Genet. 1993; 3: 283-291
        • Lilienbaum A.
        • Li Z.
        • Butler-Browne G.
        • Bolmont C.
        • Grimaud J.A.
        • Paulin D.
        Human desmin gene: utilization as a marker of human muscle differentiation.
        Cell Mol Biol. 1988; 34: 663-672
        • Kaufman S.J.
        • Foster R.F.
        Replicating myoblasts express a muscle-specific phenotype.
        Proc Natl Acad Sci USA. 1988; 85: 9606-9610
        • Li Z.
        • Marchand P.
        • Humbert J.
        • Babinet C.
        • Paulin D.
        Desmin sequence elements regulating skeletal muscle-specific expression in transgenic mice.
        Development. 1993; 117: 947-959
        • Miller J.B.
        • Stockdale F.E.
        Developmental origins of skeletal muscle fibers: clonal analysis of myogenic cell lineages based on expression of fast and slow myosin heavy chains.
        Proc Natl Acad Sci USA. 1986; 83: 3860-3864
        • Miller J.B.
        • Crow M.T.
        • Stockdale F.E.
        Slow and fast myosin heavy chain content defines three types of myotubes in early muscle cell cultures.
        J Cell Biol. 1985; 101: 1643-1650
        • Holdwe E.
        • Maeda M.
        • Bies R.D.
        Expression and regulation of the dystrophin Purkinje promoter in human skeletal muscle, heart, and brain.
        Hum Genet. 1996; 97: 232-239