Inhibition of hepadnaviral replication by polyethylenimine-based intravenous delivery of antisense phosphodiester oligodeoxynucleotides to the liver.
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.
Robaczewska
M
M
INSERM U271, Lyon, France, and Faculty of Biotechnology, University of Gdansk and Medical University of Gdansk, Gdansk, Poland.
Guerret
S
S
Remy
J S
JS
Chemin
I
I
Offensperger
W B
WB
Chevallier
M
M
Behr
J P
JP
Podhajska
A J
AJ
Blum
H E
HE
Trepo
C
C
Cova
L
L
D000818:Animals; D015316:Genetic Therapy / Q000379:methods*; D018347:Hepadnaviridae Infections / Q000628:therapy*; D015611:Hepatitis B Virus, Duck / Q000235:genetics*; D015151:Immunoblotting; D007668:Kidney / Q000378:metabolism; D008099:Liver / Q000821:virology*; D008168:Lung / Q000378:metabolism; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D023421:Models, Animal; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage*; D011094:Polyethyleneimine; D013154:Spleen / Q000378:metabolism; D014779:Virus Replication / Q000235:genetics
D016376:Oligonucleotides, Antisense; D011094:Polyethyleneimine