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Topical application of plasmid DNA to mouse and human skin

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Abstract

Gene expression following direct injection of naked plasmid DNA into the skin has been demonstrated in the past. Topical application of plasmid DNA represents an attractive route of gene delivery. If successful, it would have great prospects in skin gene therapy since it is painless and easy to apply. In this study, we analyzed the expression of plasmid DNA in vivo and in vitro following topical application of plasmid DNA in various liposomal spray formulations. Therefore, different concentrations of plasmid DNA expressing enhanced green fluorescent protein (pEGFP-N1) were sprayed onto mouse or human skin once daily for three consecutive days and compared with direct injection. Gene expression was assessed 24 h after the final topical application of various liposomal DNA formulations. The results showed that EGFP mRNA and protein were detectable by RT-PCR and Western blot, respectively. However, epicutaneously applied EGFP plasmid DNA did not lead to microscopically detectable EGFP protein, when assessed by confocal laser microscopy or fluorescence-activated cell sorting in contrast to about 4% of fluorescent keratinocytes following intradermal injection. In an in vivo mouse model, the application of pEGFP-N1 DNA led to the generation of GFP-specific antibodies. These results indicate that topical spray application of pEGFP-N1 liposomal DNA formulations is a suitable method for plasmid DNA delivery to the skin, yielding limited gene expression. This spray method may thus be useful for DNA vaccination. To increase its attractiveness for skin gene therapy, the improvement of topical formulations with enhanced DNA absorption is desirable.

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Acknowledgements

We are grateful to Dr. R. Piotrowiak, MIKA Company, Bad Oeynhausen, Germany, for providing several lots of different liposomal spray formulations. We are also grateful to Nicole C. Bartosch-Stolp for expert editorial assistance.

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Author notes

  1. Nuschin Meykadeh

    Present address: Department of Dermatology and Venerology, Martin-Luther-Universität Halle, Germany

Authors and Affiliations

  1. Department of Dermatology, Heinrich-Heine-University of Düsseldorf, Moorenstr. 5, 40225, Düsseldorf, Germany

    Nuschin Meykadeh, Alireza Mirmohammadsadegh, Etiena Basner-Tschakarjan & Ulrich R. Hengge

  2. Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY, 14853, USA

    Zhijun Wang

Authors
  1. Nuschin Meykadeh
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  2. Alireza Mirmohammadsadegh
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  3. Zhijun Wang
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  4. Etiena Basner-Tschakarjan
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  5. Ulrich R. Hengge
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Correspondence to Ulrich R. Hengge.

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Meykadeh, N., Mirmohammadsadegh, A., Wang, Z. et al. Topical application of plasmid DNA to mouse and human skin. J Mol Med 83, 897–903 (2005). https://doi.org/10.1007/s00109-005-0669-x

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  • DOI: https://doi.org/10.1007/s00109-005-0669-x

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