Electroporation Enhanced Effect of Dystrophin Splice Switching PNA Oligomers in Normal and Dystrophic Muscle

Peptide nucleic acid (PNA) is a synthetic DNA mimic that has shown potential for discovery of novel splice switching antisense drugs. However, in vivo cellular delivery has been a limiting factor for development, and only few successful studies have been reported. As a possible modality for improvem...

Descripción completa

Detalles Bibliográficos
Autores principales: Brolin, Camilla, Shiraishi, Takehiko, Hojman, Pernille, Krag, Thomas O, Nielsen, Peter E, Gehl, Julie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5014535/
https://www.ncbi.nlm.nih.gov/pubmed/26623939
http://dx.doi.org/10.1038/mtna.2015.41
_version_ 1782452306871058432
author Brolin, Camilla
Shiraishi, Takehiko
Hojman, Pernille
Krag, Thomas O
Nielsen, Peter E
Gehl, Julie
author_facet Brolin, Camilla
Shiraishi, Takehiko
Hojman, Pernille
Krag, Thomas O
Nielsen, Peter E
Gehl, Julie
author_sort Brolin, Camilla
collection PubMed
description Peptide nucleic acid (PNA) is a synthetic DNA mimic that has shown potential for discovery of novel splice switching antisense drugs. However, in vivo cellular delivery has been a limiting factor for development, and only few successful studies have been reported. As a possible modality for improvement of in vivo cellular availability, we have investigated the effect of electrotransfer upon intramuscular (i.m.) PNA administration in vivo. Antisense PNA targeting exon 23 of the murine dystrophin gene was administered by i.m. injection to the tibialis anterior (TA) muscle of normal NMRI and dystrophic mdx mice with or without electroporation. At low, single PNA doses (1.5, 3, or 10 µg/TA), electroporation augmented the antisense exon skipping induced by an unmodified PNA by twofold to fourfold in healthy mouse muscle with optimized electric parameters, measured after 7 days. The PNA splice switching was detected at the RNA level up to 4 weeks after a single-dose treatment. In dystrophic muscles of the MDX mouse, electroporation increased the number of dystrophin-positive fibers about 2.5-fold at 2 weeks after a single PNA administration compared to injection only. In conclusion, we find that electroporation can enhance PNA antisense effects in muscle tissue.
format Online
Article
Text
id pubmed-5014535
institution National Center for Biotechnology Information
language English
publishDate 2015
publisher Nature Publishing Group
record_format MEDLINE/PubMed
spelling pubmed-50145352016-09-19 Electroporation Enhanced Effect of Dystrophin Splice Switching PNA Oligomers in Normal and Dystrophic Muscle Brolin, Camilla Shiraishi, Takehiko Hojman, Pernille Krag, Thomas O Nielsen, Peter E Gehl, Julie Mol Ther Nucleic Acids Original Article Peptide nucleic acid (PNA) is a synthetic DNA mimic that has shown potential for discovery of novel splice switching antisense drugs. However, in vivo cellular delivery has been a limiting factor for development, and only few successful studies have been reported. As a possible modality for improvement of in vivo cellular availability, we have investigated the effect of electrotransfer upon intramuscular (i.m.) PNA administration in vivo. Antisense PNA targeting exon 23 of the murine dystrophin gene was administered by i.m. injection to the tibialis anterior (TA) muscle of normal NMRI and dystrophic mdx mice with or without electroporation. At low, single PNA doses (1.5, 3, or 10 µg/TA), electroporation augmented the antisense exon skipping induced by an unmodified PNA by twofold to fourfold in healthy mouse muscle with optimized electric parameters, measured after 7 days. The PNA splice switching was detected at the RNA level up to 4 weeks after a single-dose treatment. In dystrophic muscles of the MDX mouse, electroporation increased the number of dystrophin-positive fibers about 2.5-fold at 2 weeks after a single PNA administration compared to injection only. In conclusion, we find that electroporation can enhance PNA antisense effects in muscle tissue. Nature Publishing Group 2015-12 2015-12-01 /pmc/articles/PMC5014535/ /pubmed/26623939 http://dx.doi.org/10.1038/mtna.2015.41 Text en Copyright © 2015 Official journal of the American Society of Gene & Cell Therapy http://creativecommons.org/licenses/by-nc-sa/4.0/ This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/4.0/
spellingShingle Original Article
Brolin, Camilla
Shiraishi, Takehiko
Hojman, Pernille
Krag, Thomas O
Nielsen, Peter E
Gehl, Julie
Electroporation Enhanced Effect of Dystrophin Splice Switching PNA Oligomers in Normal and Dystrophic Muscle
title Electroporation Enhanced Effect of Dystrophin Splice Switching PNA Oligomers in Normal and Dystrophic Muscle
title_full Electroporation Enhanced Effect of Dystrophin Splice Switching PNA Oligomers in Normal and Dystrophic Muscle
title_fullStr Electroporation Enhanced Effect of Dystrophin Splice Switching PNA Oligomers in Normal and Dystrophic Muscle
title_full_unstemmed Electroporation Enhanced Effect of Dystrophin Splice Switching PNA Oligomers in Normal and Dystrophic Muscle
title_short Electroporation Enhanced Effect of Dystrophin Splice Switching PNA Oligomers in Normal and Dystrophic Muscle
title_sort electroporation enhanced effect of dystrophin splice switching pna oligomers in normal and dystrophic muscle
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5014535/
https://www.ncbi.nlm.nih.gov/pubmed/26623939
http://dx.doi.org/10.1038/mtna.2015.41
work_keys_str_mv AT brolincamilla electroporationenhancedeffectofdystrophinspliceswitchingpnaoligomersinnormalanddystrophicmuscle
AT shiraishitakehiko electroporationenhancedeffectofdystrophinspliceswitchingpnaoligomersinnormalanddystrophicmuscle
AT hojmanpernille electroporationenhancedeffectofdystrophinspliceswitchingpnaoligomersinnormalanddystrophicmuscle
AT kragthomaso electroporationenhancedeffectofdystrophinspliceswitchingpnaoligomersinnormalanddystrophicmuscle
AT nielsenpetere electroporationenhancedeffectofdystrophinspliceswitchingpnaoligomersinnormalanddystrophicmuscle
AT gehljulie electroporationenhancedeffectofdystrophinspliceswitchingpnaoligomersinnormalanddystrophicmuscle