Antisense Oligo-Mediated Multiple Exon Skipping in a Dog Model of Duchenne Muscular Dystrophy
Exon skipping is currently one of the most promising molecular therapies for Duchenne muscular dystrophy (DMD). We have recently developed multiple exon skipping targeting exons 6 and 8 in dystrophin mRNA of canine X-linked muscular dystrophy (CXMD), an animal model of DMD, which exhibits severe dystrophic phenotype in skeletal muscles and cardiac muscle. We have induced efficient exon skipping both in vitro and in vivo by using cocktail antisense 2’O-methyl oligonucleotides (2’OMePS) and cocktail phosphorodiamidate morpholino oligomers (morpholinos, or PMOs) and ameliorated phenotype of dystrophic dogs by systemic injections. The multiple exon skipping (double exon skipping) shown here provides the prospect of choosing deletions that optimize the functionality of the truncated dystrophin protein for DMD patients by using a common cocktail that could be validated as a single drug and also potentially applicable for more than 90% of DMD patients.
Key wordsMultiple exon skipping Morpholinos (phosphorodiamidate morpholino oligomers) 2¢O-methylated antisense oligomers (phosphorothioate) Dystrophic dogs (canine X-linked muscular dystrophy) Duchenne/Becker muscular dystrophies
Authors thank Drs. Terence Partridge, Stephanie Duguez (Children’s National Medical Center, Washington DC), Masanori Kobayashi, Yoshitsugu Aoki, Takashi Saito, Katsutoshi Yuasa, Naoko Yugeta, Sachiko Ohshima, Jin-Hong Shin, Michiko Wada, Kazuhiro Fukushima, Satoru Masuda, Kazue Kinoshita, Hideki Kita, Shin-ichi Ichikawa, Yumiko Yahata, Takayuki Nakayama, Akinori Nakamura (National Institute of Neuroscience, Tokyo, Japan), Adam Rabinowitz, and Jonathan Beauchamp (Imperial College, London, UK), Qi-long Lu (Carolinas Medical Center) for discussions and technical assistance. This work was supported by the Foundation to Eradicate Duchenne, the Department of Defense CDMRP program, the Jain Foundation, the Crystal Ball of Virginia Beach (Muscular Dystrophy Association USA), the National Center for Medical Rehabilitation Research, the NIH Wellstone Muscular Dystrophy Research Centers, and the Ministry of Health, Labor, and Welfare of Japan (Research on Nervous and Mental Disorders, 16B-2, 19A-7; Health and Labor Sciences, Research Grants for Translation Research, H19-translational research-003, Health Sciences Research Grants Research on Psychiatry and Neurological Disease and Mental Health, H18-kokoro-019).
- 6.Lu, Q. L., Rabinowitz, A., Chen, Y. C., Yokota, T., Yin, H., Alter, J., Jadoon, A., Bou-Gharios, G., and Partridge, T. (2005) Systemic delivery of antisense oligoribonucleotide restores dystrophin expression in body-wide skeletal muscles. Proc Natl Acad Sci USA 102, 198–203.PubMedCrossRefGoogle Scholar
- 8.Sharp, N. J., Kornegay, J. N., Van Camp, S. D., Herbstreith, M. H., Secore, S. L., Kettle, S., Hung, W. Y., Constantinou, C. D., Dykstra, M. J., Roses, A. D., and et al. (1992) An error in dystrophin mRNA processing in golden retriever muscular dystrophy, an animal homologue of Duchenne muscular dystrophy. Genomics 13, 115–121.PubMedCrossRefGoogle Scholar
- 11.Yokota, T., Lu, Q. L., Morgan, J. E., Davies, K. E., Fisher, R., Takeda, S., and Partridge, T. A. (2006) Expansion of revertant fibers in dystrophic mdx muscles reflects activity of muscle precursor cells and serves as an index of muscle regeneration. J Cell Sci 119, 2679–2687.PubMedCrossRefGoogle Scholar