Targeted exon skipping in transgenic hDMD mice: A model for direct preclinical screening of human-specific antisense oligonucleotides

The therapeutic potential of frame-restoring exon skipping by antisense oligonucleotides (AONs) has recently been demonstrated in cultured muscle cells from a series of Duchenne muscular dystrophy (DMD) patients. To facilitate clinical application, in vivo studies in animal models are required to de...

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Bibliographic Details
Published in:Molecular therapy 2004-08, Vol.10 (2), p.232-240
Main Authors: Bremmer-Bout, Mattie, Aartsma-Rus, Annemieke, de Meijer, Emile J, Kaman, Wendy E, Janson, Anneke A M, Vossen, Rolf H A M, van Ommen, Gert-Jan B, den Dunnen, Johan T, van Deutekom, Judith C T
Format: Article
Language:English
Subjects:
Animals
Disease Models, Animal
Drug Evaluation, Preclinical
Dystrophin - genetics
Dystrophin - metabolism
Efficiency
Exons - genetics
Gene Targeting - methods
Gene therapy
Humans
Mice
Mice, Transgenic
Muscle, Skeletal - chemistry
Muscle, Skeletal - cytology
Muscle, Skeletal - metabolism
Muscular dystrophy
Muscular Dystrophy, Duchenne - drug therapy
Muscular Dystrophy, Duchenne - genetics
Mutation
Oligonucleotides, Antisense - analysis
Oligonucleotides, Antisense - genetics
Oligonucleotides, Antisense - pharmacology
RNA, Messenger - analysis
RNA, Messenger - metabolism
Toxicity
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Summary:The therapeutic potential of frame-restoring exon skipping by antisense oligonucleotides (AONs) has recently been demonstrated in cultured muscle cells from a series of Duchenne muscular dystrophy (DMD) patients. To facilitate clinical application, in vivo studies in animal models are required to develop safe and efficient AON-delivery methods. However, since exon skipping is a sequence-specific therapy, it is desirable to target the human DMD gene directly. We therefore set up human sequence-specific exon skipping in transgenic mice carrying the full-size human gene (hDMD). We initially compared the efficiency and toxicity of intramuscular AON injections using different delivery reagents in wild-type mice. At a dose of 3.6 nmol AON and using polyethylenimine, the skipping levels accumulated up to 3% in the second week postinjection and lasted for 4 weeks. We observed a correlation of this long-term effect with the intramuscular persistence of the AON. In regenerating myofibers higher efficiencies (up to 9%) could be obtained. Finally, using the optimized protocols in hDMD mice, we were able to induce the specific skipping of human DMD exons without affecting the endogenous mouse gene. These data highlight the high sequence specificity of this therapy and present the hDMD mouse as a unique model to optimize human-specific exon skipping in vivo.
ISSN:1525-0016
1525-0024
DOI:10.1016/j.ymthe.2004.05.031