Abstract
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.
Keywords
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Article info
Publication history
Accepted:
September 30,
2004
Received in revised form:
August 18,
2004
Received:
April 20,
2004
Identification
Copyright
© 2004 Elsevier B.V. Published by Elsevier Inc. All rights reserved.
