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Inhibition of hepadnaviral replication by polyethylenimine-based intravenous delivery of antisense phosphodiester oligodeoxynucleotides to the liver

Gene Therapy volume 8, pages 874–881 (2001)Cite this article

Abstract

Antisense oligodeoxynucleotides (ODNs) appear as attractive anti-hepatitis B virus (HBV) agents. We investigated in vivo, in the duck HBV (DHBV) infection model, whether linear polyethylenimine (lPEI)-based intravenous delivery of the natural antisense phosphodiester ODNs (O-ODNs) can prevent their degradation and allow viral replication inhibition in the liver. DHBV-infected Pekin ducklings were injected with antisense O-ODNs covering the initiation codon of the DHBV large envelope protein, either in free form (O-ODN-AS2) or coupled to lPEI (lPEI/O-ODN-AS2). Following optimization of lPEI/O-ODN complex formulation, complete O-ODN condensation into a homogenous population of small (20–60 nm) spherical particles was achieved. Flow cytometry analysis showed that lPEI-mediated transfer allowed the intrahepatic delivery of lPEI/O-ODN-AS2 to increase three-fold as compared with the O-ODN-AS2. Following 9-day therapy the intrahepatic levels of both DHBV DNA and RNA were significantly decreased in the lPEI/O-ODN-AS2-treated group as compared with the O-ODN-AS2-treated, control lPEI/O-ODN-treated, and untreated controls. In addition, inhibition of intrahepatic viral replication by lPEI/O-ODN-AS2 was not associated with toxicity and was comparable with that induced by the phosphorothioate S-ODN-AS2 at a five-fold higher dose. Taken together, our results demonstrate that phosphodiester antisense lPEI/O-ODN complexes specifically inhibit hepadnaviral replication. Therefore we provide here the first in vivo evidence that intravenous treatment with antisense phosphodiester ODNs coupled to lPEI can selectively block a viral disease-causing gene in the liver.

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Acknowledgements

We thank Darius Moradpour for stimulating discussion, advice and critcal review of the manuscript. The excellent assistance of C Jamard with animals is gratefully acknowledged. MR was the recipient of fellowships from Région Rhône-Alpes (TEMPRA), French Ministère des Affaires Etrangères and Association pour la Recherche sur le Cancer (ARC). WBO and HEB acknowledge support by the Zentrum für Klinische Forschung I, University of Freiburg, Germany, and by grant Of 14/4–2 from the Deutsche Forschungsgemeinschaft. This work was supported in part by a grant from TEMPRA and from ARC.

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Authors and Affiliations

  1. INSERM U271, Lyon, France

    M Robaczewska, I Chemin, C Trepo & L Cova

  2. Faculty of Biotechnology, University of Gdansk and Medical University of Gdansk, Gdansk, Poland

    M Robaczewska & A J Podhajska

  3. Biomaterials Laboratory, Faculty of Pharmacy, Lyon, France

    S Guerret

  4. Faculty of Pharmacy, Illkirch, France

    J-S Remy & J-P Behr

  5. Department of Medicine, University of Freiburg, Germany

    W-B Offensperger & H E Blum

  6. Laboratoires Marcel Mérieux, Lyon, France

    M Chevallier

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  1. M Robaczewska
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  4. I Chemin
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Robaczewska, M., Guerret, S., Remy, JS. et al. Inhibition of hepadnaviral replication by polyethylenimine-based intravenous delivery of antisense phosphodiester oligodeoxynucleotides to the liver. Gene Ther 8, 874–881 (2001). https://doi.org/10.1038/sj.gt.3301464

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