A Dystrophin Exon-52 Deleted Miniature Pig Model of Duchenne Muscular Dystrophy and Evaluation of Exon Skipping

Yusuke Echigoya, Nhu Trieu, William Duddy, Hong M. Moulton, Haifang Yin, Terence A. Partridge, Eric P. Hoffman, Joe N. Kornegay, Frank A. Rohret, Christopher S. Rogers, Toshifumi Yokota

Research output: Contribution to journalArticlepeer-review

3 Citations (Scopus)
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Abstract

Duchenne muscular dystrophy (DMD) is a lethal X-linked recessive disorder caused by mutations in the DMD gene and the subsequent lack of dystrophin protein. Recently, phosphorodiamidate morpholino oligomer (PMO)-antisense oligonucleotides (ASOs) targeting exon 51 or 53 to reestablish the DMD reading frame have received regulatory approval as commercially available drugs. However, their applicability and efficacy remain limited to particular patients. Large animal models and exon skipping evaluation are essential to facilitate ASO development together with a deeper understanding of dystrophinopathies. Using recombinant adeno-associated virus-mediated gene targeting and somatic cell nuclear transfer, we generated a Yucatan miniature pig model of DMD with an exon 52 deletion mutation equivalent to one of the most common mutations seen in patients. Exon 52-deleted mRNA expression and dystrophin deficiency were confirmed in the skeletal and cardiac muscles of DMD pigs. Accordingly, dystrophin-associated proteins failed to be recruited to the sarcolemma. The DMD pigs manifested early disease onset with severe bodywide skeletal muscle degeneration and with poor growth accompanied by a physical abnormality, but with no obvious cardiac phenotype. We also demonstrated that in primary DMD pig skeletal muscle cells, the genetically engineered exon-52 deleted pig DMD gene enables the evaluation of exon 51 or 53 skipping with PMO and its advanced technology, peptide-conjugated PMO. The results show that the DMD pigs developed here can be an appropriate large animal model for evaluating in vivo exon skipping efficacy.
Original languageEnglish
Article number13065
Pages (from-to)1-21
Number of pages21
JournalInternational Journal of Molecular Sciences
Volume22
Issue number23
Early online date2 Dec 2021
DOIs
Publication statusPublished (in print/issue) - 2 Dec 2021

Bibliographical note

Funding Information:
This research was funded by the National Institute of Arthritis and Musculoskeletal and Skin Diseases (AR061900), Canadian Institutes of Health Research (CIHR), Women and Children?s Health Research Institute (Canada), The Friends of Garrett Cumming Research Chair Fund (Canada), Muscular Dystrophy Canada, HM Toupin Neurological Science Research Chair Fund (Canada), Canada Foundation for Innovation, Alberta Enterprise and Advanced Education (Canada), Jesse?s Journey?Foundation for Gene and Cell Therapy (Canada), University of Alberta (Canada), CIHR China?Canada Joint Health Research Initiative, JSPS Postdoctoral Fellowship for Overseas Researchers (Japan).

Funding Information:
Funding: This research was funded by the National Institute of Arthritis and Musculoskeletal and Skin Diseases (AR061900), Canadian Institutes of Health Research (CIHR), Women and Children’s Health Research Institute (Canada), The Friends of Garrett Cumming Research Chair Fund (Can‐ ada), Muscular Dystrophy Canada, HM Toupin Neurological Science Research Chair Fund (Can‐ ada), Canada Foundation for Innovation, Alberta Enterprise and Advanced Education (Canada), Jesse’s Journey–Foundation for Gene and Cell Therapy (Canada), University of Alberta (Canada), CIHR China‐Canada Joint Health Research Initiative, JSPS Postdoctoral Fellowship for Overseas Researchers (Japan).

Publisher Copyright:
© 2021 by the authors. Licensee MDPI, Basel, Switzerland.

Keywords

  • Antisense oligonucleotide
  • DMD
  • Dystrophin
  • Exon skipping
  • Large animal model
  • Morpholino
  • Pig model

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