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Pluronic–PEI copolymers enhance exon-skipping of 2′-O-methyl phosphorothioate oligonucleotide in cell culture and dystrophic mdx mice

Gene Therapy volume 21, pages 52–59 (2014)Cite this article

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Abstract

A series of small-size polyethylenimine (PEI)-conjugated pluronic polycarbamates (PCMs) have been investigated for the ability to modulate the delivery of 2′-O-methyl phosphorothioate RNA (2′-OMePS) in vitro and in dystrophic mdx mice. The PCMs retain strong binding capacity to negatively charged oligomer as demonstrated by agarose gel retardation assay, with the formation of condensed polymer/oligomer complexes at a wide-range weight ratio from 1:1 to 20:1. The condensed polymer/oligomer complexes form 100–300 nm nanoparticles. Exon-skipping effect of 2′-OMePS was dramatically enhanced with the use of the most effective PCMs in comparison with 2′-OMePS alone in both cell culture and in vivo, respectively. More importantly, the effective PCMs, especially those composed of moderate size (2k–5kDa) and intermediate hydrophilic–lipophilic balance (7–23) of pluronics, enhanced exon-skipping of 2′-OMePS with low toxicity as compared with Lipofectamine-2000 in vitro or PEI 25k in vivo. The variability of individual PCM for delivery of antisense oligomer and plasmid DNA indicate the complexity of interaction between polymer and their cargos. Our data demonstrate the potential of PCMs to mediate delivery of modified antisense oligonucleotides to the muscle for treating muscular dystrophy or other appropriate myodegenerative diseases.

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Acknowledgements

We thank Dr Craig A Ogle at the Department of Chemistry, University of North Carolina, Charlotte, for guidance in measuring 1H-NMR and DLS. We also thank Ms Daisy Ridings and Mr David Radoff for TEM analysis, Dr David M Foureau for fluorescence-activated cell sorting analysis. We also gratefully acknowledge the Carolinas Muscular Dystrophy Research Endowment at the Carolinas HealthCare Foundation and Carolinas Medical Center, Charlotte, NC, USA.

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Authors and Affiliations

  1. Department of Neurology, McColl Lockwood Laboratory for Muscular Dystrophy Research, Neuromuscular/ALS Center, Carolinas Medical Center, Charlotte, NC, USA

    M Wang, B Wu, P Lu, J D Tucker, S Milazi, S N Shah & Q L Lu

  2. Department of Biology, University of North Carolina, Charlotte, NC, USA

    J D Tucker

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  1. M Wang
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Correspondence to M Wang or Q L Lu.

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Wang, M., Wu, B., Lu, P. et al. Pluronic–PEI copolymers enhance exon-skipping of 2′-O-methyl phosphorothioate oligonucleotide in cell culture and dystrophic mdx mice. Gene Ther 21, 52–59 (2014). https://doi.org/10.1038/gt.2013.57

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