Sequence-specific inhibition of duck hepatitis B virus reverse transcription by peptide nucleic acids (PNA)

• Published in: Journal of hepatology Vol. 42; no. 2; pp. 180 - 187
• Main Authors: Robaczewska, Magdalena, Narayan, Ramamurthy, Seigneres, Beatrice, Schorr, Olivier, Thermet, Alexandre, Podhajska, Anna J., Trepo, Christian, Zoulim, Fabien, Nielsen, Peter E., Cova, Lucyna
• Format: Journal Article
• Language: English
• Published: Oxford Elsevier B.V 01.02.2005; Elsevier
• Subjects: Animals; Base Sequence; Biological and medical sciences; DNA Primers; Ducks; Embryo, Nonmammalian; Hepatitis B virus; Hepatitis B Virus, Duck - enzymology; Hepatitis B Virus, Duck - genetics; Hepatocyte; Human viral diseases; Infectious diseases; Medical sciences; Molecular Sequence Data; Oligodeoxyribonucleotides - pharmacology; Peptide nucleic acid (PNA); Peptide Nucleic Acids - pharmacology; Reverse transcription; Reverse Transcription - genetics; RNA-Directed DNA Polymerase - metabolism; Viral diseases; Viral diseases of the digestive system; Viral hepatitis; PNA; RT; DHBV; Reverse transcription; Peptide nucleic acid (PNA); Hepatocyte; ε; O-ODN; PDH; ODN; S-ODN; Hepatitis B virus; HBV; Duck hepatitis B virus; Peptides; Orthohepadnavirus; Hepatic disease; Infection; Virus; Viral hepatitis B; Avihepadnavirus; Hepadnaviridae; Viral disease; Digestive diseases; Inhibition; Nucleic acid
• ISSN: 0168-8278; 1600-0641
• DOI: 10.1016/j.jhep.2004.10.010

Peptide nucleic acids (PNAs) appear as promising new antisense agents, that have not yet been examined as hepatitis B virus (HBV) inhibitors. Our aim was to study the ability of PNAs targeting the duck HBV (DHBV) encapsidation signal ε to inhibit reverse transcription (RT) and to compare their efficacy with phosphorothioate oligodeoxynucleotides (S-ODNs). The effect of two partly overlapping PNAs targeting ε and of analogous S-ODNs was tested in cell-free transcription and translation system for DHBV RT expression. In addition their antiviral effect was investigated in primary duck hepatocytes (PDH). Both PNAs reproducibly inhibited DHBV RT in a dose-dependent manner with IC 50 of 10 nM, whereas up to 600-fold higher concentration of S-ODNs was required for similar inhibition. The PNA targeting the bulge and upper stem of ε appeared as more efficient RT inhibitor than the PNA targeting only the bulge. Importantly, the inhibition was highly sequence-specific since double-mismatched PNA had no effect on the RT reaction. Moreover, in PDH the PNA coupled to Arg 7 cationic delivery peptide decreased DHBV replication. We provide the first evidence that PNAs targeting the bulge and upper stem of ε can efficiently and in a sequence-specific manner inhibit DHBV RT.