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

TY - JOUR

T1 - Sequence-specific inhibition of duck hepatitis B virus reverse transcription by peptide nucleic acids (PNA)

AU - Robaczewska, Magdalena

AU - Narayan, Ramamurthy

AU - Seigneres, Beatrice

AU - Schorr, Olivier

AU - Thermet, Alexandre

AU - Podhajska, Anna J

AU - Trepo, Christian

AU - Zoulim, Fabien

AU - Nielsen, Peter E

AU - Cova, Lucyna

PY - 2005/2

Y1 - 2005/2

N2 - BACKGROUND/AIMS: 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 epsilon to inhibit reverse transcription (RT) and to compare their efficacy with phosphorothioate oligodeoxynucleotides (S-ODNs).METHODS: The effect of two partly overlapping PNAs targeting epsilon 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).RESULTS: Both PNAs reproducibly inhibited DHBV RT in a dose-dependent manner with IC(50) of 10nM, whereas up to 600-fold higher concentration of S-ODNs was required for similar inhibition. The PNA targeting the bulge and upper stem of epsilon 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.CONCLUSIONS: We provide the first evidence that PNAs targeting the bulge and upper stem of epsilon can efficiently and in a sequence-specific manner inhibit DHBV RT.

AB - BACKGROUND/AIMS: 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 epsilon to inhibit reverse transcription (RT) and to compare their efficacy with phosphorothioate oligodeoxynucleotides (S-ODNs).METHODS: The effect of two partly overlapping PNAs targeting epsilon 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).RESULTS: Both PNAs reproducibly inhibited DHBV RT in a dose-dependent manner with IC(50) of 10nM, whereas up to 600-fold higher concentration of S-ODNs was required for similar inhibition. The PNA targeting the bulge and upper stem of epsilon 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.CONCLUSIONS: We provide the first evidence that PNAs targeting the bulge and upper stem of epsilon can efficiently and in a sequence-specific manner inhibit DHBV RT.

KW - Animals

KW - Base Sequence

KW - DNA Primers

KW - Ducks

KW - Embryo, Nonmammalian

KW - Hepatitis B Virus, Duck

KW - Molecular Sequence Data

KW - Oligodeoxyribonucleotides

KW - Peptide Nucleic Acids

KW - RNA-Directed DNA Polymerase

KW - Reverse Transcription

U2 - 10.1016/j.jhep.2004.10.010

DO - 10.1016/j.jhep.2004.10.010

M3 - Journal article

C2 - 15664242

SN - 0168-8278

VL - 42

SP - 180

EP - 187

JO - Journal of Hepatology

JF - Journal of Hepatology

IS - 2

ER -