Nucleoside/nucleotide analogues are widely used to treat chronic HBV infection, but drug resistance is common. The role of HBV surface gene variants in drug resistance to nucleoside/nucleotide analogues is unknown. We are trying to investigate the dynamics of S gene mutations and how they relate to a patient's virological response in this study.
Methods
Thirty patients with chronic hepatitis B were enrolled and serum samples were collected at multiple time points during treatment with telbivudine (LdT). The coding regions of the small surface antigen (S-HBsAg) were amplified and sequenced using the 454 GS FLX+ System.
Results
Sequencing results revealed different dynamics of non-synonymous mutations, such as sL9P, sN40S, sG44E, sW172*, sW182* and sS187F, between patients with a complete virological response and those with a partial virological response. The viral population heterogeneity decreased at week 12 of LdT treatment in patients with a complete virological response, with a concomitant decline in non-synonymous mutations (from an average of 14 to 9.9 per sample) and an increase in the frequencies of major variants (from 14.3% to 40.4%).
Conclusions
Our findings suggest that the decrease in viral population heterogeneity at an early stage of LdT treatment was associated with the subsequent optimal virological response, and the early appearance of some specific mutations, such as sG44E, sW172* and sW182*, is a potential indicator of a partial virological response in continuing therapy.
References
1.
WhalleyS.A., MurrayJ.M., BrownD.Kinetics of acute hepatitis B virus infection in humans.J Exp Med2001; 193: 847–854.
2.
OsiowyC., GilesE., TanakaY., MizokamiM., MinukG.Y.Molecular evolution of hepatitis B virus over 25 years.J Virol2006; 80: 10307–10314.
3.
DrakeJ.W., CharlesworthB., CharlesworthD., CrowJ.F.Rates of spontaneous mutation.Genetics1998; 148: 1667–1686.
4.
NowakM.A., BonhoefferS., HillA.M., BoehmeR., ThomasH.C., McDadeH.Viral dynamics in hepatitis B virus infection.Proc Natl Acad Sci U S A1996; 93: 4398–4402.
5.
LaiC.L., LeungN., TeoE.K.A 1-year trial of telbivudine, lamivudine, and the combination in patients with hepatitis B e antigen-positive chronic hepatitis B.Gastroenterology2005; 129: 528–536.
6.
FungS.K., AndreoneP., HanS.H.Adefovir-resistant hepatitis B can be associated with viral rebound and hepatic decompensation.J Hepatol2005; 43: 937–943.
7.
AllenM.I., DeslauriersM., AndrewsC.W.Identification and characterization of mutations in hepatitis B virus resistant to lamivudine. Lamivudine clinical investigation group.Hepatology1998; 27: 1670–1677.
8.
AngusP., VaughanR., XiongS.Resistance to adefovir dipivoxil therapy associated with the selection of a novel mutation in the HBV polymerase.Gastroenterology2003; 125: 292–297.
9.
WarnerN., LocarniniS.Mechanisms of hepatitis B virus resistance development.Intervirology2014; 57: 218–224.
10.
CarmanW F.. The clinical significance of surface antigen variants of hepatitis B virus.J Viral Hepat1997; 4Suppl 1: 11–20.
11.
ChotiyaputtaW., LokA.S.Hepatitis B virus variants.Nat Rev Gastroenterol Hepatol2009; 6: 453–462.
12.
YamaniL.N., YanoY., UtsumiT.Ultradeep sequencing for detection of quasispecies variants in the major hydrophilic region of hepatitis B virus in Indonesian patients.J Clin Microbiol2015; 53: 3165–3175.
13.
YouH., JiaJ.Telbivudine treatment in chronic hepatitis B: experience from China.J Viral Hepat2013; 20Suppl 1: 3–8.
14.
HouJ., YinY.K., XuD.Telbivudine versus lamivudine in Chinese patients with chronic hepatitis B: results at 1 year of a randomized, double-blind trial.Hepatology2008; 47: 447–454.
15.
ZhouB., DongH., HeY.Composition and interactions of hepatitis B virus quasispecies defined the virological response during telbivudine therapy.Sci Rep2015; 5: 17123.
16.
SunJ., XieQ., TanD.The 104-week efficacy and safety of telbivudine-based optimization strategy in chronic hepatitis B patients: a randomized, controlled study.Hepatology2014; 59: 1283–1292.
17.
ZhouB., XiaoL., WangZ., ChangE.T., ChenJ., HouJ.Geographical and ethnic distribution of the HBV C/D recombinant on the Qinghai-Tibet Plateau.PLoS ONE2011; 6: e18708.
18.
LiH., DurbinR.Fast and accurate long-read alignment with Burrows-Wheeler transform.Bioinformatics2010; 26: 589–595.
19.
LiH.A statistical framework for SNP calling, mutation discovery, association mapping and population genetical parameter estimation from sequencing data.Bioinformatics2011; 27: 2987–2993.
20.
Margeridon-ThermetS., ShulmanN.S., AhmedA.Ultra-deep pyrosequencing of hepatitis B virus quasispecies from nucleoside and nucleotide reverse-transcriptase inhibitor (NRTI)-treated patients and NRTI-naive patients.J Infect Dis2009; 199: 1275–1285.
21.
YuenM.F., YuanH.J., WongD.K.Prognostic determinants for chronic hepatitis B in Asians: therapeutic implications.Gut2005; 54: 1610–1614.
22.
DingH., LiuB., ZhaoC.Amino acid similarities and divergences in the small surface proteins of genotype C hepatitis B viruses between nucleos(t)ide analogue-naive and lamivudine-treated patients with chronic hepatitis B.Antiviral Res2014; 102: 29–34.
23.
YehC.T., ChienR.N., ChuC.M., LiawY.F.Clearance of the original hepatitis B virus YMDD-motif mutants with emergence of distinct lamivudine-resistant mutants during prolonged lamivudine therapy.Hepatology2000; 31: 1318–1326.
24.
WakilS.M., KazimS.N., KhanL.A.Prevalence and profile of mutations associated with lamivudine therapy in Indian patients with chronic hepatitis B in the surface and polymerase genes of hepatitis B virus.J Med Virol2002; 68: 311–318.
25.
LocarniniS.Primary resistance, multidrug resistance, and cross-resistance pathways in HBV as a consequence of treatment failure.Hepatol Int2008; 2: 147–151.
26.
YangH., WestlandC.E., DelaneyW.E.4thResistance surveillance in chronic hepatitis B patients treated with adefovir dipivoxil for up to 60 weeks.Hepatology2002; 36: 464–473.
27.
SheldonJ., CaminoN., RodesB.Selection of hepatitis B virus polymerase mutations in HIV-coinfected patients treated with tenofovir.Antivir Ther2005; 10: 727–734.
28.
AhnS.H., ParkY.K., ParkE.S.The impact of the hepatitis B virus polymerase rtA181T mutation on replication and drug resistance is potentially affected by overlapping changes in surface gene.J Virol2014; 88: 6805–6818.
29.
WarnerN., LocarniniS.The antiviral drug selected hepatitis B virus rtA181T/sW172* mutant has a dominant negative secretion defect and alters the typical profile of viral rebound.Hepatology2008; 48: 88–98.
30.
KweiK., TangX., LokA.S.Impaired virion secretion by hepatitis B virus immune escape mutants and its rescue by wild-type envelope proteins or a second-site mutation.J Virol2013; 87: 2352–2357.
31.
GaoZ.Y., LiT., WangJ.Mutations in preS genes of genotype C hepatitis B virus in patients with chronic hepatitis B and hepatocellular carcinoma.J Gastroenterol2007; 42: 761–768.
32.
GuidottiL.G., ChisariF.V.Immunobiology and pathogenesis of viral hepatitis.Annu Rev Pathol2006; 1: 23–61.
33.
WielandS.F., ChisariF.V.Stealth and cunning: hepatitis B and hepatitis C viruses.J Virol2005; 79: 9369–9380.
34.
ChenY., LiX., YeB.Effect of telbivudine therapy on the cellular immune response in chronic hepatitis B.Antiviral Res2011; 91: 23–31.
35.
ShiT.D., ZhangJ.M., WangX.F.Effects of antiviral therapy with telbivudine on peripheral iNKT cells in HBeAg(+) chronic hepatitis B patients.Clin Exp Med2012; 12: 105–113.
36.
EvansA., RivaA., CooksleyH.Programmed death 1 expression during antiviral treatment of chronic hepatitis B: impact of hepatitis B e-antigen seroconversion.Hepatology2008; 48: 759–769.
37.
JiangX., ZhangM., LaiQ.Restored circulating invariant NKT cells are associated with viral control in patients with chronic hepatitis B.PLoS ONE2011; 6: e28871.
38.
WuZ.G., YanW.M., GuoW.Telbivudine preserves T-helper 1 cytokine production and downregulates programmed death ligand 1 in a mouse model of viral hepatitis.J Viral Hepat2010; 17Suppl 1: 24–33.
39.
NishijimaN., MarusawaH., UedaY.Dynamics of hepatitis B virus quasispecies in association with nucleos(t)ide analogue treatment determined by ultra-deep sequencing.PLoS ONE2012; 7: e35052.
40.
ChenL., ZhangQ., YuD.M., WanM.B., ZhangX.X.Early changes of hepatitis B virus quasispecies during lamivudine treatment and the correlation with antiviral efficacy.J Hepatol2009; 50: 895–905.
41.
WeberB.Genetic variability of the S gene of hepatitis B virus: clinical and diagnostic impact.J Clin Virol2005; 32: 102–112.
42.
EdwardsL.J., EvavoldB.D.T cell recognition of weak ligands: roles of signaling, receptor number, and affinity.Immunol Res2011; 50: 39–48.
