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

E. G. Duysen, K. Parikh, V. Aleti, V. Manne, Oksana Lockridge, N. Chilukuri

Research output: Contribution to journalArticle

27 Citations (Scopus)

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 2LD 50 doses of diazoxon. Four of the six mice survived a second dose of diazoxon (for a total of 4LD 50) administered 24 h later. In contrast, none of the three mice in the control group survived one 2LD 50 dose. These results show that hPON1 in mice functions as a prophylactic and offers significant protection against lethal doses of diazoxon.

Original languageEnglish (US)
Pages (from-to)250-257
Number of pages8
JournalGene Therapy
Volume18
Issue number3
DOIs
StatePublished - Mar 1 2011

Fingerprint

Human Adenoviruses
Pesticides
Genes
Adenoviridae
HDL Lipoproteins
Chemical Warfare Agents
diazoxon
Culture Media
Cell Culture Techniques
Molecular Weight
Western Blotting
Control Groups

Keywords

  • adenovirus
  • chemical warfare nerve agents
  • gene delivery
  • organophosphorus pesticides
  • paraoxonase1

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

Adenovirus-mediated human paraoxonase1 gene transfer to provide protection against the toxicity of the organophosphorus pesticide toxicant diazoxon. / Duysen, E. G.; Parikh, K.; Aleti, V.; Manne, V.; Lockridge, Oksana; Chilukuri, N.

In: Gene Therapy, Vol. 18, No. 3, 01.03.2011, p. 250-257.

Research output: Contribution to journalArticle

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