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Adenovirus-mediated human paraoxonase1 gene transfer to provide protection against the toxicity of the organophosphorus pesticide toxicant diazoxon

Gene Therapy volume 18, pages 250–257 (2011)Cite this article

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Abstract

Human paraoxonase1 (hPON1) is a potential therapeutic against the toxicity of organophosphorus (OP) pesticides and chemical warfare nerve agents. We tested whether PON1 gene transfer using adenovirus provides protection against the toxicity of the OP diazoxon. Using an adenovirus construct containing hPON1 gene, we showed elevated levels of recombinant hPON1 in vitro in 293A cells and in vivo in mice. The recombinant enzyme was secreted by 293A cells into culture medium and into the systemic circulation of mice. Western blotting revealed that the virally expressed hPON1 had the expected molecular weight of 45 kDa. Recombinant hPON1 in mice was in complex with mouse high-density lipoprotein (HDL) and migrated more slowly than endogenous hPON1 in the human HDL complex. Mice injected with adenovirus expressed PON1 at 600–3480 U ml–1 on day 5 post-treatment, which is 8–50-fold above endogenous. Six mice expressing hPON1 survived 2LD50 doses of diazoxon. Four of the six mice survived a second dose of diazoxon (for a total of 4LD50) administered 24 h later. In contrast, none of the three mice in the control group survived one 2LD50 dose. These results show that hPON1 in mice functions as a prophylactic and offers significant protection against lethal doses of diazoxon.

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Acknowledgements

This work was supported by funds from Defense Threat Reduction Agency (DTRA), Department of Defense (Project No. 1.D003_09_WR_C to Dr Chilukuri) and US Army Medical and Materiel Command W81XWH-07-2-0034 (to Dr Oksana Lockridge). We also thank Dr Gregory E Garcia (Division of Biochemistry, WRAIR) for valuable suggestions with the manuscript.

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  1. E G Duysen and K Parikh: These authors contributed equally to this work.

Authors and Affiliations

  1. Eppley Institute, University of Nebraska Medical Center, Omaha, NE, USA

    E G Duysen & O Lockridge

  2. Division of Biochemistry, Department of Molecular Pharmacology, Walter Reed Army Institute of Research, 503 Robert Grant Avenue, Silver Spring, MD, USA,

    K Parikh, V Aleti, V Manne & N Chilukuri

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  1. E G Duysen
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Correspondence to N Chilukuri.

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The opinions or assertions contained herein are the private views of the authors and are not to be construed as official or as reflecting the views of the Army or the Department of Defense, USA.

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Duysen, E., Parikh, K., Aleti, V. et al. Adenovirus-mediated human paraoxonase1 gene transfer to provide protection against the toxicity of the organophosphorus pesticide toxicant diazoxon. Gene Ther 18, 250–257 (2011). https://doi.org/10.1038/gt.2010.136

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