Aminoglycoside antibiotics restore dystrophin function to skeletal muscles of mdx mice
Elisabeth R. Barton-Davis, … , Stuart E. Leland, H. Lee Sweeney
Elisabeth R. Barton-Davis, … , Stuart E. Leland, H. Lee Sweeney
Published August 15, 1999
Citation Information: J Clin Invest. 1999;104(4):375-381. https://doi.org/10.1172/JCI7866.
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Article

Aminoglycoside antibiotics restore dystrophin function to skeletal muscles of mdx mice

Abstract

Duchenne muscular dystrophy (DMD) is caused by mutations in the dystrophin gene, leading to the absence of the dystrophin protein in striated muscle. A significant number of these mutations are premature stop codons. On the basis of the observation that aminoglycoside treatment can suppress stop codons in cultured cells, we tested the effect of gentamicin on cultured muscle cells from the mdx mouse — an animal model for DMD that possesses a premature stop codon in the dystrophin gene. Exposure of mdx myotubes to gentamicin led to the expression and localization of dystrophin to the cell membrane. We then evaluated the effects of differing dosages of gentamicin on expression and functional protection of the muscles of mdx mice. We identified a treatment regimen that resulted in the presence of dystrophin in the cell membrane in all striated muscles examined and that provided functional protection against muscular injury. To our knowledge, our results are the first to demonstrate that aminoglycosides can suppress stop codons not only in vitro but also in vivo. Furthermore, these results raise the possibility of a novel treatment regimen for muscular dystrophy and other diseases caused by premature stop codon mutations. This treatment could prove effective in up to 15% of patients with DMD.

Authors

Elisabeth R. Barton-Davis, Laurence Cordier, Daria I. Shoturma, Stuart E. Leland, H. Lee Sweeney

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The effect of gentamicin treatment on eccentric contraction injury. Effe...
The effect of gentamicin treatment on eccentric contraction injury. Effective treatment regimes are depicted in a. The decrement in force after 5 eccentric contractions was significantly less in mdx EDL muscles injected with gentamicin (mdxGi200, n = 8; 200% dose equivalent) compared with untreated mdx EDLs (mdxctrl, n = 8) and muscles from Becker construct (22) transgenic animals (Beck tg, n = 5). The level of protection afforded by gentamicin in mdx muscles approached the values measured in EDLs from untreated C57 mice (C57ctrl, n = 4), and was not significantly different from EDLs from gentamicin-treated C57 mice (C57Gi200, n = 4). DHB treatment, when combined with 200% gentamicin (mdxGi200D, n = 4), did not alter levels of protection by this gentamicin dose alone. Lower injected doses of gentamicin (50% dose equivalent) were very effective at preventing force decrements in female mdx mice (mdxGi50F, n = 4). Treatment regimens that were not effective in preventing eccentric contractile injury in male mdx mice are displayed in b. These include 400% gentamicin injected in combination with DHB (mdxGi400D, n = 4), 100% injected gentamicin (mdxGi100, n = 4), and all mdx animals treated by infusion pumps (mdxGP50, 50% dose; mdxGP100, 100% dose; mdxGP200, 200% dose; n = 4 for each condition). The measurements of force generation after eccentric contraction were reflected in the measurements of membrane damage (c). There was a significant reduction in the proportion of fibers with dye entry in EDLs from treated mdx mice compared with muscles from untreated mdx muscles. P < 0.05 for comparisons between treated mdx muscles and untreated C57 controls. *P < 0.05 for comparisons between treated and untreated mdx muscles.