Journal article
Immortalized canine dystrophic myoblast cell lines for development of peptide-conjugated splice-switching oligonucleotides
- Abstract:
- Duchenne muscular dystrophy (DMD) is a severe muscle-wasting disease caused by frameshift or nonsense mutations in the DMD gene, resulting in the loss of dystrophin from muscle membranes. Exon skipping using splice-switching oligonucleotides (SSOs) restores the reading frame of DMD pre-mRNA by generating internally truncated but functional dystrophin protein. To potentiate effective tissue-specific targeting by functional SSOs, it is essential to perform accelerated and reliable in vitro screening-based assessment of novel oligonucleotides and drug delivery technologies, such as cell-penetrating peptides, before their in vivo pharmacokinetic and toxicity evaluation. We have established novel canine immortalized myoblast lines by transducing murine cyclin-dependent kinase-4 and human telomerase reverse transcriptase genes into myoblasts isolated from beagle-based wild-type or canine X-linked muscular dystrophy in Japan (CXMDJ) dogs. These myoblast lines exhibited improved myogenic differentiation and increased proliferation rates compared with passage-15 primary parental myoblasts, and their potential to differentiate into myotubes was maintained in later passages. Using these dystrophin-deficient immortalized myoblast lines, we demonstrate that a novel cell-penetrating peptide (Pip8b2)-conjugated SSO markedly improved multiexon skipping activity compared with the respective naked phosphorodiamidate morpholino oligomers. In vitro screening using immortalized canine cell lines will provide a basis for further pharmacological studies on drug delivery tools.
- Publication status:
- Published
- Peer review status:
- Peer reviewed
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(Preview, Version of record, 750.4KB, Terms of use)
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- Publisher copy:
- 10.1089/nat.2020.0907
Authors
- Publisher:
- Mary Ann Liebert
- Journal:
- Nucleic Acid Therapeutics More from this journal
- Volume:
- 31
- Issue:
- 2
- Pages:
- 172-181
- Publication date:
- 2021-02-09
- Acceptance date:
- 2020-12-24
- DOI:
- EISSN:
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2159-3345
- ISSN:
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2159-3337
- Pmid:
-
33567244
- Language:
-
English
- Keywords:
- Pubs id:
-
1161325
- Local pid:
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pubs:1161325
- Deposit date:
-
2021-06-18
Terms of use
- Copyright holder:
- Tone et al.
- Copyright date:
- 2021
- Rights statement:
- ©2021 Yuichiro Tone, et al. Published by Mary Ann Liebert, Inc. This Open Access article is distributed under the terms of the Creative Commons License [CC-BY] ( http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
- Licence:
- CC Attribution (CC BY)
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