In Vitro Models for Studying Hepatitis B Virus Drug Resistance

ABSTRACT

Hepatitis B virus resistance to antivirals is conferred by the selection of polymerase gene mutants with reduced susceptibility to nucleoside analogues. Because of the increasing number of resistant mutants to the different available drugs, several phenotypic assays have been developed to provide cross-resistance data to guide clinicians in the management of therapy. Their possible use in the routine follow-up of antiviral treatment is also discussed. The different in vitro models for the study of HBV resistance include enzymatic and cell-based assays. Their relevance for the analysis of viral fitness, drug susceptibility, and cross-resistance is also reviewed.

KEYWORDS

Hepatitis B virus - replication - drug resistance - nucleoside analogues

REFERENCES

• 1 Zoulim F. Mechanism of viral persistence and resistance to nucleoside and nucleotide analogs in chronic hepatitis B virus infection.  Antiviral Res. 2004;  64 1-15
• 2 Locarnini S, Hatzakis A, Heathcote J et al.. Management of antiviral resistance in patients with chronic hepatitis B.  Antivir Ther. 2004;  9 679-693
• 3 Xiong X, Flores C, Yang H, Toole J J, Gibbs C S. Mutations in hepatitis B DNA polymerase associated with resistance to lamivudine do not confer resistance to adefovir in vitro.  Hepatology. 1998;  28 1669-1673
• 4 Xiong X, Yang H, Westland C E, Zou R, Gibbs C S. In vitro evaluation of hepatitis B virus polymerase mutations associated with famciclovir resistance.  Hepatology. 2000;  31 219-224
• 5 Seigneres B, Pichoud C, Ahmed S S, Hantz O, Trepo C, Zoulim F. Evolution of hepatitis B virus polymerase gene sequence during famciclovir therapy for chronic hepatitis B.  J Infect Dis. 2000;  181 1221-1233
• 6 Pichoud C, Seignères B, Wang Z, Trepo C, Zoulim F. Transient selection of a hepatitis B virus polymerase gene mutant associated with a decreased replication capacity and famciclovir resistance.  Hepatology. 1999;  29 230-237
• 7 Fu L, Liu S H, Cheng Y C. Sensitivity of L-(-)2,3-dideoxythiacytidine resistant hepatitis B virus to other antiviral nucleoside analogues.  Biochem Pharmacol. 1999;  57 1351-1359
• 8 Fu L, Cheng Y-C. Role of additional mutations outside the YMDD motif of hepatitis B virus polymerase in L(-)SddC (3TC) resistance.  Biochem Pharmacol. 1998;  55 1567-1572
• 9 Aye T T, Bartholomeusz A, Shaw T et al.. Hepatitis B virus polymerase mutations during antiviral therapy in a patient following liver transplantation.  J Hepatol. 1997;  26 1148-1153
• 10 Tenney D J, Levine S M, Rose R E et al.. Clinical emergence of entecavir-resistant hepatitis B virus requires additional substitutions in virus already resistant to Lamivudine.  Antimicrob Agents Chemother. 2004;  48 3498-3507
• 11 Wang G H, Seeger C. The reverse transcriptase of hepatitis B virus acts a protein primer for viral DNA synthesis.  Cell. 1992;  71 663-670
• 12 Zoulim F, Dannaoui E, Borel C et al.. 2′,3′-dideoxy-b-L-5-fluorocytidine inhibits duck hepatitis B virus reverse transcription and suppresses viral DNA synthesis in hepatocytes, both in vitro and in vivo.  Antimicrob Agents Chemother. 1996;  40 448-453
• 13 Staschke K, Colacino J. Priming of duck hepatitis B virus reverse transcription in vitro: premature termination of primer DNA induced by the 5′-triphosphate of fialuridine.  J Virol. 1994;  68 8265-8269
• 14 Le Guerhier F, Pichoud C, Guerret S et al.. Characterization of the antiviral effect of 2′,3′-dideoxy-2′, 3′- didehydro-beta-L-5-fluorocytidine in the duck hepatitis B virus infection model.  Antimicrob Agents Chemother. 2000;  44 111-122
• 15 Seigneres B, Pichoud C, Martin P, Furman P, Trepo C, Zoulim F. Inhibitory activity of dioxolane purine analogs on wild-type and lamivudine-resistant mutants of hepadnaviruses.  Hepatology. 2002;  36 710-722
• 16 Seigneres B, Aguesse-Germon S, Pichoud C et al.. Duck hepatitis B virus polymerase gene mutants associated with resistance to lamivudine have a decreased replication capacity in vitro and in vivo.  J Hepatol. 2001;  34 114-122
• 17 Brunelle M N, Jacquard A C, Pichoud C et al.. Susceptibility to antivirals of a human HBV strain with mutations conferring resistance to both lamivudine and adefovir.  Hepatology. 2005;  41 1391-1398
• 18 Jacquard A C, Brunelle M N, Pichoud C et al.. In vitro characterization of the anti-hepatitis B virus activity and cross-resistance profile of 2′,3′-dideoxy-3′-fluoroguanosine.  Antimicrob Agents Chemother. 2006;  50 955-961
• 19 Melegari M, Scaglioni P P, Wands J R. Hepatitis B virus mutants associated with 3TC and famciclovir administration are replication defective.  Hepatology. 1998;  27 628-633
• 20 Allen M I, Deslauriers M, Andrews C W et al.. Identification and characterization of mutations in hepatitis B virus resistant to Lamivudine.  Hepatology. 1998;  27 1670-1677
• 21 Gunther S, Li B C, Miska S, Kruger D H, Meisel H, Will H. A novel method for efficient amplification of whole hepatitis B virus genomes permits rapid functional analysis and reveals deletion mutants in immunosuppressed patients.  J Virol. 1995;  69 5437-5444
• 22 Villeneuve J P, Durantel D, Durantel S et al.. Selection of a hepatitis B virus strain resistant to adefovir in a liver transplantation patient.  J Hepatol. 2003;  39 1085-1089
• 23 Durantel D, Carrouee-Durantel S, Werle-Lapostolle B et al.. A new strategy for studying in vitro the drug susceptibility of clinical isolates of human hepatitis B virus.  Hepatology. 2004;  40 855-864
• 24 Lada O, Benhamou Y, Cahour A, Katlama C, Poynard T, Thibault V. In vitro susceptibility of lamivudine-resistant hepatitis B virus to adefovir and tenofovir.  Antivir Ther. 2004;  9 353-363
• 25 Benhamou Y, Tubiana R, Thibault V. Tenofovir disoproxil fumarate in patients with HIV and lamivudine-resistant hepatitis B virus.  N Engl J Med. 2003;  348 177-178
• 26 Dore G J, Cooper D A, Pozniak A L et al.. Efficacy of tenofovir disoproxil fumarate in antiretroviral therapy-naive and-experienced patients coinfected with HIV-1 and hepatitis B virus.  J Infect Dis. 2004;  189 1185-1192
• 27 van Bommel F, Wunsche T, Mauss S et al.. Comparison of adefovir and tenofovir in the treatment of lamivudine-resistant hepatitis B virus infection.  Hepatology. 2004;  40 1421-1425
• 28 Lacombe K, Gozlan J, Boelle P Y et al.. Long-term hepatitis B virus dynamics in HIV-hepatitis B virus-co-infected patients treated with tenofovir disoproxil fumarate.  AIDS. 2005;  19 907-915
• 29 Angus P, Vaughan R, Xiong S et al.. Resistance to adefovir dipivoxil therapy associated with the selection of a novel mutation in the HBV polymerase.  Gastroenterology. 2003;  125 292-297
• 30 Sheldon J, Camino N, Rodes B et al.. Selection of hepatitis B virus polymerase mutations in HIV-coinfected patients treated with tenofovir.  Antivir Ther. 2005;  10 727-734
• 31 Yang H, Westland C, Xiong S, Delaney W E. In vitro antiviral susceptibility of full-length clinical hepatitis B virus isolates cloned with a novel expression vector.  Antiviral Res. 2004;  61 27-36
• 32 Ladner S, Miller T J, King R W. The M539V polymerase variant of human hepatitis B virus demonstrates resistance to 2′-deoxy-3′-thiacytidine and a reduced ability to synthesize viral DNA.  Antimicrob Agents Chemother. 1998;  42 2128-2131
• 33 Fu L, Cheng Y C. Characterization of novel human hepatoma cell lines with stable hepatitis B virus secretion for evaluating new compounds against lamivudine- and penciclovir-resistant virus.  Antimicrob Agents Chemother. 2000;  44 3402-3407
• 34 Delaney W E, Miller T G, Isom H C. Use of the hepatitis B virus recombinant baculovirus-HepG2 system to study the effects of (-)-beta-2′,3′-dideoxy-3′-thiacytidine on replication of hepatitis B virus and accumulation of covalently closed circular DNA.  Antimicrob Agents Chemother. 1999;  43 2017-2026
• 35 Delaney W E, Edwards R, Colledge D et al.. Cross-resistance testing of antihepadnaviral compounds using novel recombinant baculoviruses which encode drug-resistant strains of hepatitis B virus.  Antimicrob Agents Chemother. 2001;  45 1705-1713
• 36 Harrison T J. Hepatitis B virus: molecular virology and common mutants.  Semin Liver Dis. 2006;  26 87-96
• 37 Bartholomew M M, Jansen R W, Jeffers L J et al.. Hepatitis-B-virus resistance to lamivudine given for recurrent infection after orthotopic liver transplantation.  Lancet. 1997;  349 20-22
• 38 Gripon P, Rumin S, Urban S et al.. Infection of a human hepatoma cell line by hepatitis B virus.  Proc Natl Acad Sci USA. 2002;  99 15655-15660
• 39 Urban S, Gripon P. Inhibition of duck hepatitis B virus infection by a myristoylated pre-S peptide of the large viral surface protein.  J Virol. 2002;  76 1986-1990
• 40 Glebe D, Urban S, Knoop E V et al.. Mapping of the hepatitis B virus attachment site by use of infection-inhibiting preS1 lipopeptides and Tupaia hepatocytes.  Gastroenterology. 2005;  129 234-245
• 41 Delaney W E, Edwards R, Colledge D et al.. Phenylpropylamine derivatives AT-61 and AT-130 inhibit replication of wild-type and lamivudine-resistant strains of hepatitis B virus in vitro.  Antimicrob Agents Chemother. 2002;  46 3057-3060
• 42 Ying C, De Clercq E, Nicholson W, Furman P, Neyts J. Inhibition of the replication of the DNA polymerase M550V mutation variant of human hepatitis B virus by adefovir, tenofovir, L-FMAU, DAPD, penciclovir and lobucavir.  J Viral Hepat. 2000;  7 161-165
• 43 Ying C, Holy A, Hockova D, Havlas Z, De Clercq E, Neyts J. Novel acyclic nucleoside phosphonate analogues with potent anti-hepatitis B virus activities.  Antimicrob Agents Chemother. 2005;  49 1177-1180
• 44 Yang H, Qi X, Sabogal A, Miller M, Xiong S, Delaney W E. Cross-resistance testing of next-generation nucleoside and nucleotide analogues against lamivudine-resistant HBV.  Antivir Ther. 2005;  10 625-633
• 45 Stuyver L, Van Geyt C, De Gendt S et al.. Line probe assay for monitoring drug resistance in hepatitis B virus-infected patients during antiviral therapy.  J Clin Microbiol. 2000;  38 702-707
• 46 Nafa S, Ahmed S, Tavan D et al.. Early detection of viral resistance by determination of hepatitis B virus polymerase mutations in patients treated by lamivudine for chronic hepatitis B.  Hepatology. 2000;  32 1078-1088
• 47 Aberle S W, Kletzmayr J, Watschinger B, Schmied B, Vetter N, Puchhammer-Stockl E. Comparison of sequence analysis and the INNO-LiPA HBV DR line probe assay for detection of lamivudine-resistant hepatitis B virus strains in patients under various clinical conditions.  J Clin Microbiol. 2001;  39 1972-1974
• 48 Clavel F, Hance A J. HIV drug resistance.  N Engl J Med. 2004;  350 1023-1035
• 49 Hanna G J, D'Aquila R T. Clinical use of genotypic and phenotypic drug resistance testing to monitor antiretroviral chemotherapy.  Clin Infect Dis. 2001;  32 774-782
• 50 Garcia-Lerma J G, Heneine W. Resistance of human immunodeficiency virus type 1 to reverse transcriptase and protease inhibitors: genotypic and phenotypic testing.  J Clin Virol. 2001;  21 197-212