Skip to main content

Advertisement

Log in

Hepatitis B virus genotype C encoding resistance mutations that emerge during adefovir dipivoxil therapy: in vitro replication phenotype

  • Original Article
  • Published:
Hepatology International Aims and scope Submit manuscript

Abstract

Introduction

Hepatitis B virus (HBV) can be classified into ten genotypes (A–J), with genotypes B and C being the most common in Asia. Recent data suggest that the HBV genotype can influence disease progression, and genotype C has been associated with more aggressive liver disease than that of other genotypes. Although there is a preventative vaccine, chronic infection remains a public health problem with oral nucleos(t)ide analog therapy being the most common treatment. The HBV genome is composed of four partially overlapping reading frames, meaning that substitutions in the HBV polymerase selected during NA therapy may also alter the overlapping HBV surface antigen (HBsAg). We have recently shown that for HBV genotype D, the rtA181T/sW172stop substitution conferring resistance to adefovir dipivoxil (ADV) alters secretion of HBsAg and exerts a dominant-negative effect on wild-type virion secretion. However, the effect of this and other ADV-resistance-associated mutations on HBV replication and HBsAg secretion for the HBV genotype C, the genotype with the most severe clinical prognosis, is unknown.

Methods/Results

We constructed 1.2-mer infectious cDNA clones of HBV genotype C encoding mutations associated with ADV resistance and established an in vitro replication assay in Huh7 cells. Decreased levels of HBV DNA and HBsAg were detected for all ADV variants relative to the 1.2-mer wild-type polymerase control plasmid. Importantly, less HBsAg was detected in the cells transfected with the rtA181T resistance mutants, and the overlapping sW172stop mutation ablated secretion of HBsAg into cell culture supernatants.

Conclusions

The identification of secretion-defective HBV in the setting of ADV therapy for HBV genotype C, and to a lesser extent HBV genotype B, has major implications for the diagnosis and treatment of HBV in the Asia-Pacific region, as it is likely that quantitative HBsAg and viral load testing of serum from patients infected with HBV encoding rtA181T and rtN236T substitutions may not accurately reflect the level of replication within hepatocytes.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Lavanchy D. Hepatitis B virus epidemiology, disease burden, treatment, and current and emerging prevention and control measures. J Viral Hepat 2004;11(2):97–107

    Article  CAS  Google Scholar 

  2. Lupberger J, Hildt E. Hepatitis B virus-induced oncogenesis. World J Gastroenterol 2007;13(1):74–81

    Article  CAS  Google Scholar 

  3. Parkin DM. Global cancer statistics in the year 2000. Lancet Oncol 2001;2(9):533–543

    Article  CAS  Google Scholar 

  4. Ayoub WS, Keeffe EB. Review article: current antiviral therapy of chronic hepatitis B. Aliment Pharmacol Ther 2011;34(10):1145–1158

    Article  CAS  Google Scholar 

  5. 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(2):292–297

    Article  Google Scholar 

  6. Zoulim F, Locarnini S. Hepatitis B virus resistance to nucleos(t)ide analogues. Gastroenterology 2009;137(5):1593–1608. e1–e2

    Article  CAS  Google Scholar 

  7. Zoulim F, Locarnini S. Management of treatment failure in chronic hepatitis B. J Hepatol 2012;56(Suppl 1):S112–S122

    Article  CAS  Google Scholar 

  8. Warner N, Locarnini S. The antiviral drug selected hepatitis B virus rtA181T/sW172* mutant has a dominant negative secretion defect and alters the typical profile of viral rebound. Hepatology 2008;48(1):88–98

    Article  CAS  Google Scholar 

  9. Yeh CT, Chen T, Hsu CW, et al. Emergence of the rtA181T/sW172* mutant increased the risk of hepatoma occurrence in patients with lamivudine-resistant chronic hepatitis B. BMC Cancer 2011;11:398

    Article  CAS  Google Scholar 

  10. Lai MW, Yeh CT. The oncogenic potential of hepatitis B virus rtA181T/surface truncation mutant. Antivir Ther 2008;13(7):875–879

    CAS  PubMed  Google Scholar 

  11. Kim BK, Revill PA, Ahn SH. HBV genotypes: relevance to natural history, pathogenesis and treatment of chronic hepatitis B. Antivir Ther 2011;16(8):1169–1186

    Article  CAS  Google Scholar 

  12. Lin CL, Kao JH. The clinical implications of hepatitis B virus genotype: recent advances. J Gastroenterol Hepatol 2011;26(Suppl 1):123–130

    Article  Google Scholar 

  13. Warner N, Locarnini S, Kuiper M, et al. The L80I substitution in the reverse transcriptase domain of the hepatitis B virus polymerase is associated with lamivudine resistance and enhanced viral replication in vitro. Antimicrob Agents Chemother 2007;51(7):2285–2292. doi:https://doi.org/10.1128/AAC.01499-06..

    Article  CAS  Google Scholar 

  14. Yuen KWL, Ayres A, Littlejohn M, et al. SEQHEPB: a sequence and analysis program and relational database system for chronic hepatitis B antiviral research. Antivir Res 2007;75:64–74

    Article  CAS  Google Scholar 

  15. Chen RY, Edwards R, Shaw T, et al. Effect of the G1896A precore mutation on drug sensitivity and replication yield of lamivudine-resistant HBV in vitro. Hepatology 2003;37(1):27–35

    Article  CAS  Google Scholar 

  16. Thompson AJ, Nguyen T, Iser D, et al. Serum hepatitis B surface antigen and hepatitis B e antigen titers: disease phase influences correlation with viral load and intrahepatic hepatitis B virus markers. Hepatology 2010;51(6):1933–1944

    Article  CAS  Google Scholar 

  17. Delaney WEt, 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 2010;45(6):1705–1713

    Article  Google Scholar 

  18. Chan HL, Hui AY, Wong ML, et al. Genotype C hepatitis B virus infection is associated with an increased risk of hepatocellular carcinoma. Gut 2004;53(10):1494–1498

    Article  Google Scholar 

  19. Chan HL, Wong ML, Hui AY, et al. Hepatitis B virus genotype C takes a more aggressive disease course than hepatitis B virus genotype B in hepatitis B e antigen-positive patients. J Clin Microbiol 2003;41(3):1277–1279

    Article  Google Scholar 

  20. Chu CJ, Hussain M, Lok AS. Hepatitis B virus genotype B is associated with earlier HBeAg seroconversion compared with hepatitis B virus genotype C. Gastroenterology 2002;122(7):1756–1762

    Article  CAS  Google Scholar 

  21. Liu CJ, Kao JH. Genetic variability of hepatitis B virus and response to antiviral therapy. Antivir Ther 2008;13(5):613–624

    CAS  PubMed  Google Scholar 

  22. Wiegand J, Hasenclever D, Tillmann HL. Should treatment of hepatitis B depend on hepatitis B virus genotypes? A hypothesis generated from an explorative analysis of published evidence. Antivir Ther 2008;13(2):211–220

    CAS  PubMed  Google Scholar 

  23. Trojan J, Stuermer M, Teuber G, Berger A, Faust D. Treatment of patients with lamivudine-resistant and adefovir dipivoxil-resistant chronic hepatitis B virus infection: is tenofovir the answer? Gut 2007;56(3):436–437

    Article  CAS  Google Scholar 

  24. Patterson SJ, George J, Strasser SI, et al. Tenofovir disoproxil fumarate rescue therapy following failure of both lamivudine and adefovir dipivoxil in chronic hepatitis B. Gut 2011;60(2):247–254

    Article  CAS  Google Scholar 

  25. Li X, Wang L, Zhong Y, et al. Hepatitis B virus (HBV) subgenotypes C2 and B2 differ in lamivudine- and adefovir-resistance-associated mutational patterns in HBV-infected Chinese patients. J Clin Microbiol 2010;48(12):4363–4369

    Article  Google Scholar 

  26. Damerow H, Yuen L, Wiegand J, et al. Mutation pattern of lamivudine resistance in relation to hepatitis B genotypes: hepatitis B genotypes differ in their lamivudine resistance associated mutation pattern. J Med Virol 2010;82(11):1850–1858

    Article  CAS  Google Scholar 

  27. Yang JX, Liu BM, Li XG, et al. Profile of HBV antiviral resistance mutations with distinct evolutionary pathways against nucleoside/nucleotide analogue treatment among Chinese chronic hepatitis B patients. Antivir Ther 2010;15(8):1171–1178

    Article  CAS  Google Scholar 

  28. Yeh CT. Development of HBV S gene mutants in chronic hepatitis B patients receiving nucleotide/nucleoside analogue therapy. Antivir Ther 2010; 15(3 Pt B):471–475

    Article  CAS  Google Scholar 

  29. Lai MW, Huang SF, Hsu CW, et al. Identification of nonsense mutations in hepatitis B virus S gene in patients with hepatocellular carcinoma developed after lamivudine therapy. Antivir Ther 2009;14(2):249–261

    CAS  PubMed  Google Scholar 

  30. Liaw Y, Kao J, Piratvisuth T, et al. Asian-Pacific Consensus statement on the management of chronic hepatitis B: A 2012 update. Hepatology Int 2012. doi:https://doi.org/10.1007/s12072-012-9365-4.

    Article  Google Scholar 

  31. Locarnini S, Bowden S. Hepatitis B surface antigen quantification: not what it seems on the surface. Hepatology 2012;56(2):411–414

    Article  CAS  Google Scholar 

  32. Kim JH, Jung YK, Joo MK, et al. Hepatitis B viral surface mutations in patients with adefovir resistant chronic hepatitis B with A181T/V polymerase mutations. J Korean Med Sci 2010;25(2):257–264

    Article  CAS  Google Scholar 

  33. Lee SA, Kim K, Kim H, Kim BJ. Nucleotide change of codon 182 in the surface gene of hepatitis B virus genotype C leading to truncated surface protein is associated with progression of liver diseases. J Hepatol 2012;56(1):63–69

    Article  CAS  Google Scholar 

Download references

Acknowledgements

WL was supported by the State Scholarship Fund from the China Scholarship Council (no. 2009601107) and a grant from the Major Science and Technology Special Project of China Eleventh 5-year Plan (no. 2008ZX10002-004). NW was supported by NHMRC project grant no. 1011024. The authors thank Scott Bowden for critically reading the manuscript.

Author information

Authors and Affiliations

Authors

Corresponding authors

Correspondence to Hui Zhuang or Peter A. Revill.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Li, W., Warner, N., Sozzi, V. et al. Hepatitis B virus genotype C encoding resistance mutations that emerge during adefovir dipivoxil therapy: in vitro replication phenotype. Hepatol Int 7, 443–450 (2013). https://doi.org/10.1007/s12072-012-9411-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12072-012-9411-2

Keywords

Navigation