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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References
Thomas CE, Ehrhardt A, Kay MA . Progress and problems with the use of viral vectors for gene therapy. Nat Rev Genet 2003; 4: 346–358.
Lentz TB, Gray SJ, Samulski RJ . Viral vectors for gene delivery to the central nervous system. Neurobiol Dis 2012; 48: 179–188.
Liu G, Li D, Pasumarthy MK, Kowalczyk TH, Gedeon CR, Hyatt SL et al. Nanoparticles of compacted DNA transfect postmitotic cells. J Biol Chem 2003; 278: 32578–32586.
Farjo R, Skaggs J, Quiambao AB, Cooper MJ, Naash MI . Efficient non-viral ocular gene transfer with compacted DNA nanoparticles. PLoS One 2006; 1: e38.
Ziady AG, Gedeon CR, Miller T, Quan W, Payne JM, Hyatt SL et al. Transfection of airway epithelium by stable PEGylated poly-L-lysine DNA nanoparticles in vivo. Mol Ther 2003; 8: 936–947.
Yurek DM, Fletcher AM, Smith GM, Seroogy KB, Ziady AG, Molter J et al. Long-term transgene expression in the central nervous system using DNA nanoparticles. Mol Ther 2009; 17: 641–650.
Fink TL, Klepcyk PJ, Oette SM, Gedeon CR, Hyatt SL, Kowalczyk TH et al. Plasmid size up to 20 kbp does not limit effective in vivo lung gene transfer using compacted DNA nanoparticles. Gene Therapy 2006; 13: 1048–1051.
Fletcher AM, Kowalczyk TH, Padegimas L, Cooper MJ, Yurek DM . Transgene expression in the striatum following intracerebral injections of DNA nanoparticles encoding for human glial cell line-derived neurotrophic factor. Neuroscience 2011; 194: 220–226.
Han Z, Conley SM, Makkia R, Guo J, Cooper MJ, Naash MI . Comparative analysis of DNA nanoparticles and AAVs for ocular gene delivery. PLoS One 2012; 7: e52189.
Padegimas L, Kowalczyk TH, Adams S, Gedeon CR, Oette SM, Dines K et al. Optimization of hCFTR lung expression in mice using DNA nanoparticles. Mol Ther 2012; 20: 63–72.
Yurek DM, Fletcher AM, McShane M, Kowalczyk TH, Padegimas L, Weatherspoon MR et al. DNA nanoparticles: detection of long-term transgene activity in brain using bioluminescence imaging. Mol Imaging 2011; 10: 327–339.
Cai X, Conley SM, Nash Z, Fliesler SJ, Cooper MJ, Naash MI . Gene delivery to mitotic and postmitotic photoreceptors via compacted DNA nanoparticles results in improved phenotype in a mouse model of retinitis pigmentosa. FASEB J 2010; 24: 1178–1191.
Han Z, Conley SM, Makkia RS, Cooper MJ, Naash MI . DNA nanoparticle-mediated ABCA4 delivery rescues Stargardt dystrophy in mice. J Clin Invest 2012; 122: 3221–3226.
Ziady AG, Gedeon CR, Muhammad O, Stillwell V, Oette SM, Fink TL et al. Minimal toxicity of stabilized compacted DNA nanoparticles in the murine lung. Mol Ther 2003; 8: 948–956.
Ding XQ, Quiambao AB, Fitzgerald JB, Cooper MJ, Conley SM, Naash MI . Ocular delivery of compacted DNA-nanoparticles does not elicit toxicity in the mouse retina. PLoS One 2009; 4: e7410.
Konstan MW, Davis PB, Wagener JS, Hilliard KA, Stern RC, Milgram LJ et al. Compacted DNA nanoparticles administered to the nasal mucosa of cystic fibrosis subjects are safe and demonstrate partial to complete cystic fibrosis transmembrane regulator reconstitution. Hum Gene Ther 2004; 15: 1255–1269.
Dhuria SV, Hanson LR, Frey 2nd WH . Intranasal delivery to the central nervous system: mechanisms and experimental considerations. J Pharm Sci 2010; 99: 1654–1673.
Wang P, Olbricht WL . Fluid mechanics in the perivascular space. J Theor Biol 2011; 274: 52–57.
Hadaczek P, Yamashita Y, Mirek H, Tamas L, Bohn MC, Noble C et al. The "perivascular pump" driven by arterial pulsation is a powerful mechanism for the distribution of therapeutic molecules within the brain. Mol Ther 2006; 14: 69–78.
Thorne RG, Pronk GJ, Padmanabhan V, Frey 2nd WH . Delivery of insulin-like growth factor-I to the rat brain and spinal cord along olfactory and trigeminal pathways following intranasal administration. Neuroscience 2004; 127: 481–496.
Rennels ML, Blaumanis OR, Grady PA . Rapid solute transport throughout the brain via paravascular fluid pathways. Adv Neurol 1990; 52: 431–439.
Lochhead JJ, Thorne RG . Intranasal delivery of biologics to the central nervous system. Adv Drug Deliv Rev 2012; 64: 614–628.
Migliore MM, Vyas TK, Campbell RB, Amiji MM, Waszczak BL . Brain delivery of proteins by the intranasal route of administration: a comparison of cationic liposomes versus aqueous solution formulations. J Pharm Sci 2010; 99: 1745–1761.
Liu XF, Fawcett JR, Thorne RG, DeFor TA, Frey 2nd WH . Intranasal administration of insulin-like growth factor-I bypasses the blood-brain barrier and protects against focal cerebral ischemic damage. J Neurol Sci 2001; 187: 91–97.
Thorne RG, Hanson LR, Ross TM, Tung D, Frey 2nd WH . Delivery of interferon-beta to the monkey nervous system following intranasal administration. Neuroscience 2008; 152: 785–797.
Kim ID, Shin JH, Kim SW, Choi S, Ahn J, Han PL et al. Intranasal delivery of HMGB1 siRNA confers target gene knockdown and robust neuroprotection in the postischemic brain. Mol Ther 2012; 20: 829–839.
Renner DB, Frey 2nd WH, Hanson LR . Intranasal delivery of siRNA to the olfactory bulbs of mice via the olfactory nerve pathway. Neurosci Lett 2012; 513: 193–197.
Lee ST, Chu K, Jung KH, Kim JH, Huh JY, Yoon H et al. miR-206 regulates brain-derived neurotrophic factor in Alzheimer disease model. Ann Neurol 2012; 72: 269–277.
Lemiale F, Kong WP, Akyurek LM, Ling X, Huang Y, Chakrabarti BK et al. Enhanced mucosal immunoglobulin A response of intranasal adenoviral vector human immunodeficiency virus vaccine and localization in the central nervous system. J Virol 2003; 77: 10078–10087.
Zhang J, Wu X, Qin C, Qi J, Ma S, Zhang H et al. A novel recombinant adeno-associated virus vaccine reduces behavioral impairment and beta-amyloid plaques in a mouse model of Alzheimer's disease. Neurobiol Dis 2003; 14: 365–379.
Laing JM, Gober MD, Golembewski EK, Thompson SM, Gyure KA, Yarowsky PJ et al. Intranasal administration of the growth-compromised HSV-2 vector DeltaRR prevents kainate-induced seizures and neuronal loss in rats and mice. Mol Ther 2006; 13: 870–881.
Draghia R, Caillaud C, Manicom R, Pavirani A, Kahn A, Poenaru L . Gene delivery into the central nervous system by nasal instillation in rats. Gene Therapy 1995; 2: 418–423.
Jiang Y, Wei N, Zhu J, Zhai D, Wu L, Chen M et al. A new approach with less damage: intranasal delivery of tetracycline-inducible replication-defective herpes simplex virus type-1 vector to brain. Neuroscience 2012; 201: 96–104.
Damjanovic D, Zhang X, Mu J, Medina MF, Xing Z . Organ distribution of transgene expression following intranasal mucosal delivery of recombinant replication-defective adenovirus gene transfer vector. Genet Vaccines Ther 2008; 6: 5.
Danielyan L, Schafer R, von Ameln-Mayerhofer A, Buadze M, Geisler J, Klopfer T et al. Intranasal delivery of cells to the brain. Eur J Cell Biol 2009; 88: 315–324.
Reitz M, Demestre M, Sedlacik J, Meissner H, Fiehler J, Kim SU et al. Intranasal delivery of neural stem/progenitor cells: a noninvasive passage to target intracerebral glioma. Stem Cells Transl Med 2012; 1: 866–873.
Gray SJ, Foti SB, Schwartz JW, Bachaboina L, Taylor-Blake B, Coleman J et al. Optimizing promoters for recombinant adeno-associated virus-mediated gene expression in the peripheral and central nervous system using self-complementary vectors. Hum Gene Ther 2011; 22: 1143–1153.
Thorne RG, Frey 2nd WH . Delivery of neurotrophic factors to the central nervous system: pharmacokinetic considerations. Clin Pharmacokinet 2001; 40: 907–946.
Han IK, Kim MY, Byun HM, Hwang TS, Kim JM, Hwang KW et al. Enhanced brain targeting efficiency of intranasally administered plasmid DNA: an alternative route for brain gene therapy. J Mol Med 2007; 85: 75–83.
Patel T, Zhou J, Piepmeier JM, Saltzman WM . Polymeric nanoparticles for drug delivery to the central nervous system. Adv Drug Deliv Rev 2012; 64: 701–705.
Wong HL, Wu XY, Bendayan R . Nanotechnological advances for the delivery of CNS therapeutics. Adv Drug Deliv Rev 2012; 64: 686–700.
Sa-Pereira I, Brites D, Brito MA . Neurovascular unit: a focus on pericytes. Mol Neurobiol 2012; 45: 327–347.
Dore-Duffy P . Pericytes: pluripotent cells of the blood brain barrier. Curr Pharm Des 2008; 14: 1581–1593.
Shimizu F, Sano Y, Abe MA, Maeda T, Ohtsuki S, Terasaki T et al. Peripheral nerve pericytes modify the blood-nerve barrier function and tight junctional molecules through the secretion of various soluble factors. J Cell Physiol 2011; 226: 255–266.
Shimizu F, Sano Y, Saito K, Abe MA, Maeda T, Haruki H et al. Pericyte-derived glial cell line-derived neurotrophic factor increase the expression of claudin-5 in the blood-brain barrier and the blood-nerve barrier. Neurochem Res 2012; 37: 401–409.
Lange S, Trost A, Tempfer H, Bauer HC, Bauer H, Rohde E et al. Brain pericyte plasticity as a potential drug target in CNS repair. Drug Discov Today 2013; 18: 456–463.
Tanudji M, Hevi S, Chuck SL . Improperly folded green fluorescent protein is secreted via a non-classical pathway. J Cell Sci 2002; 115 (Pt 19): 3849–3857.
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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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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DOI: https://doi.org/10.1038/gt.2014.28
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