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Intranasal administration of plasmid DNA nanoparticles yields successful transfection and expression of a reporter protein in rat brain

Abstract

Viral vectors are a commonly used method for gene therapy because of their highly efficient transduction of cells. However, many vectors have a small genetic capacity, and their potential for immunogenicity can limit their usefulness. Moreover, for disorders of the central nervous system (CNS), the need for invasive surgical delivery of viruses to the brain also detracts from their clinical applicability. Here, we show that intranasal delivery of unimolecularly compacted DNA nanoparticles (DNA NPs), which consist of single molecules of plasmid DNA encoding enhanced green fluorescent protein (eGFP) compacted with 10 kDa polyethylene glycol (PEG)-substituted lysine 30-mers (CK30PEG10k), successfully transfect cells in the rat brain. Direct eGFP fluorescence microscopy, eGFP-immunohistochemistry (IHC) and eGFP-ELISA all demonstrated eGFP protein expression 2 days after intranasal delivery. eGFP-positive cells were found throughout the rostral-caudal axis of the brain, most often adjacent to capillary endothelial cells. This localization provides evidence for distribution of the nasally administered DNA NPs via perivascular flow. These results are the first report that intranasal delivery of DNA NPs can bypass the blood–brain barrier and transfect and express the encoded protein in the rat brain, affording a non-invasive approach for gene therapy of CNS disorders.

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Acknowledgements

We gratefully acknowledge the assistance of Dr Justin Manjourides, biostatistician in the Department of Health Sciences at Northeastern University, for advice on methods of statistical analysis and interpretation. We also thank Amanda Nadeau for technical assistance in counting eGFP-positive cells. Brendan Harmon was supported by an IGERT Nanomedicine Science & Technology Award NSF-DGE-0965843 to Northeastern University. Funding for this research was provided in part by a Northeastern University 2011-2012 Provost's Tier 1 Interdisciplinary Grant and by the Michael J. Fox Foundation for Parkinson's Research.

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Correspondence to B L Waszczak.

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LP, OSL and MJC are employed by Copernicus Therapeutics.

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Harmon, B., Aly, A., Padegimas, L. et al. Intranasal administration of plasmid DNA nanoparticles yields successful transfection and expression of a reporter protein in rat brain. Gene Ther 21, 514–521 (2014). https://doi.org/10.1038/gt.2014.28

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