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Functional amounts of dystrophin produced by skipping the mutated exon in the mdx dystrophic mouse

Nature Medicine volume 9, pages 1009–1014 (2003)Cite this article

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Abstract

As a target for gene therapy, Duchenne muscular dystrophy (DMD) presents many obstacles but also an unparalleled prospect for correction by alternative splicing. The majority of mutations in the dystrophin gene occur in the region encoding the spectrin-like central rod domain, which is largely dispensable. Thus, splicing around mutations can generate a shortened but in-frame transcript, permitting translation of a partially functional dystrophin protein. We have tested this idea in vivo in the mdx dystrophic mouse (carrying a mutation in exon 23 of the dystrophin gene) by combining a potent transfection protocol with a 2-O-methylated phosphorothioated antisense oligoribonucleotide (2OMeAO) designed to promote skipping of the mutated exon*. The treated mice show persistent production of dystrophin at normal levels in large numbers of muscle fibers and show functional improvement of the treated muscle. Repeated administration enhances dystrophin expression without eliciting immune responses. Our data establishes the realistic practicality of an approach that is applicable, in principle, to a majority of cases of severe dystrophinopathy.

*NOTE: In the version of this article initially published online, the following errors appeared: The fourth sentence of the abstract was incorrect. It has been changed to, "We have tested this idea in vivo in the mdx dystrophic mouse (carrying a mutation in exon 23 of the dystrophin gene) by combining…to promote skipping of the mutated exon." The sixth sentence of the first paragraph of the Results was incorrect and has been changed. It now reads, "…(C57Bl/10ScSn; designated C57)…" Figure 2 and its legend contained an error. The figure should be labeled with subpanels a and b. The legend should read, "Figure 2 Detection of dystrophin and dystrophin protein complexes. Exon mapping of 2OMeAO-induced dystrophin (a,b) and detection of dystrophin protein complexes (b) in serial sections..." These mistakes have been corrected for the HTML and print versions of the article.

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Figure 1: Induction of dystrophin by 20MeAO and F127 in mice of various ages.
Figure 2: Detection of dystrophin and dystrophin protein complexes.
Figure 3: Detection of membrane-localized nNOS in fibers expressing dystrophin.
Figure 4: Duration of dystrophin expression in tibialis anterior muscles of 4-week-old mdx mice 1, 2, 4, 8 and 12 weeks after injection of 2OMeAO or sense oligonucleotide (control).
Figure 5: Physiological analysis of tibialis anterior muscles from C57 and mdx mice treated with 2OMeAO (mdx-T) or sense (mdx-C) oligonucleotides.
Figure 6: Dystrophin re-expression and lack of immune response.

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  • 16 July 2003

    The full text and PDF were updated as per note

Notes

  1. NOTE: In the version of this article initially published online, the following errors appeared: The fourth sentence of the abstract was incorrect. It has been changed to, "We have tested this idea in vivo in the mdx dystrophic mouse (carrying a mutation in exon 23 of the dystrophin gene) by combining…to promote skipping of the mutated exon." The sixth sentence of the first paragraph of the Results was incorrect and has been changed. It now reads, "…(C57Bl/10ScSn; designated C57)…" Figure 2 and its legend contained an error. The figure should be labeled with subpanels a and b. The legend should read, "Figure 2 Detection of dystrophin and dystrophin protein complexes. Exon mapping of 2OMeAO-induced dystrophin (a,b) and detection of dystrophin protein complexes (b) in serial sections..." These mistakes have been corrected for the HTML and print versions of the article.

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Acknowledgements

This work was supported by the Medical Research Council of Great Britain, the Muscular Dystrophy Group of Great Britain, the Leopold Muller Foundation, the Muscular Dystrophy Association of the USA and Western Australia, the Neuromuscular Foundation of Western Australia, and the National Health & Medical Research Council of Australia. We also thank R.C. Woledge (Institute of Human Performance, University College London) and A. Wilson (Structure and Motion Laboratory, Royal Veterinary College, London) for providing equipment for part of the muscle contraction tests.

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Authors and Affiliations

  1. Muscle Cell Biology, MRC Clinical Science Centre, Hammersmith Hospital, Du Cane Road, London, W12 0NN, UK

    Qi Long Lu, George Bou-Gharios & Terence A Partridge

  2. Australian Neuromuscular Research Institute, Centre for Neuromuscular and Neurological Disorders, QEII Medical Centre University of Western Australia, Nedlands, 6009, Western Australia

    Christopher J Mann, Sue Fletcher & Stephen D Wilton

  3. Department of Biosciences, University of Hertfordshire, College Lane, Hatfield, AL10 9AB, Herts, UK

    Fang Lou

  4. MRIC Biochemistry Group, The North East Wales Institute, Plas Coch, Mold Road, Wrexham, LL11 2AW, UK

    Glenn E Morris

  5. Department of Immunology, Division of Medicine, Imperial College, Hammersmith Hospital Campus, Du Cane Road, London, W12 0NN, UK

    Shao-an Xue

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Correspondence to Qi Long Lu or Terence A Partridge.

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Lu, Q., Mann, C., Lou, F. et al. Functional amounts of dystrophin produced by skipping the mutated exon in the mdx dystrophic mouse. Nat Med 9, 1009–1014 (2003). https://doi.org/10.1038/nm897

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