Putative and pretreatment drug resistance mutations in reverse transcriptase gene among untreated chronic hepatitis B patients at Arifin Achmad Regional District Hospital, Riau, Indonesia
Abstract
Background: Mutations within the hepatitis B virus (HBV) reverse transcriptase (RT) gene have been associated with drug resistance against nucleos(t)ide analogs (NAs).
Objective: This study aimed to identify mutations in the RT gene among patients with chronic hepatitis B (CHB) before receiving antiviral therapy and its relationship with the HBV genotypes.
Methods: A total of 26 HBV DNA was extracted from the blood plasma of CHB patients. HBV RT gene was amplified and sequenced using the Sanger dideoxy sequencing method. The HBV genotype was determined through phylogenetic analysis using the Maximum Likelihood method.
Results: The study subjects comprised 14 CHB patients without complications and 12 CHB patients with cirrhosis/hepatoma. CHB patients with cirrhosis/hepatoma were older than those without complications. The HBV genotypes comprised 15 (57.7%) genotype C and 11 (42.3%) genotype B. All treatment-naïve CHB patients did not demonstrate any classical NA resistance mutations within the RT gene. However, several putative and pretreatment resistance mutations, including F221Y, N238H, and V224I, were high frequency in more than 40% of study subjects. In addition, F221Y and N238H/Q mutations were frequently observed in genotype B, while V224 I was only found in patients infected with genotype C (p=0.000).
Conclusions: There was no evidence of classical RT gene mutations associated with NA resistance in treatment-naïve patients with CHB. However, several putative and pretreatment mutations were identified as genotype-specific mutations and may contribute to antiviral resistance against NAs.
References
World Health Organization. Global Hepatitis Report, 2017. World Health Organization; 2017.
Chen Y, Tian Z. HBV-Induced Immune Imbalance in the Development of HCC. Front Immunol. 2019;10: 2048. https://doi.org/10.3389/fimmu.2019.02048
Muljono DH. Epidemiology of hepatitis B and C in republic of indonesia. Euroasian J Hepatogastroenterol. 2017;7: 55-59. https://doi.org/10.5005/jp-journals-10018-1212
Xia Y, Liang TJ. Development of Direct-acting Antiviral and Host-targeting Agents for Treatment of Hepatitis B Virus Infection. Gastroenterology. 2019;156: 311-324. https://doi.org/10.1053/j.gastro.2018.07.057
Paraskevis D, Angelis K, Magiorkinis G, Kostaki E, Ho SYW, Hatzakis A. Dating the origin of hepatitis B virus reveals higher substitution rate and adaptation on the branch leading to F/H genotypes. Mol Phylogenet Evol. 2015;93: 44-54. https://doi.org/10.1016/j.ympev.2015.07.010
Spitz N, Mello FCA, Moreira AS, Gusatti CS, Martins RMB, Gomes SA, et al. Reconstruction of the spatial and temporal dynamics of hepatitis B virus genotype D in the Americas. PLoS One. 2019;14: e0220342. https://doi.org/10.1371/journal.pone.0220342
Yamani LN, Yano Y, Utsumi T, Wasityastuti W, Rinonce HT, Widasari DI, et al. Profile of Mutations in the Reverse Transcriptase and Overlapping Surface Genes of Hepatitis B Virus (HBV) in Treatment-Naïve Indonesian HBV Carriers. Jpn J Infect Dis. 2017;70: 647-655. https://doi.org/10.7883/yoken.JJID.2017.078
Colombatto P, Coco B, Bonino F, Brunetto MR. Management and Treatment of Patients with Chronic Hepatitis B: Towards Personalized Medicine. Viruses. 2022;14. https://doi.org/10.3390/v14040701
Allweiss L, Dandri M. The Role of cccDNA in HBV Maintenance. Viruses. 2017;9. https://doi.org/10.3390/v9060156
Trépo C, Chan HLY, Lok A. Hepatitis B virus infection. Lancet. 2014;384: 2053-2063. https://doi.org/10.1016/S0140-6736(14)60220-8
Choi Y-M, Lee S-Y, Kim B-J. Naturally occurring hepatitis B virus reverse transcriptase mutations related to potential antiviral drug resistance and liver disease progression. World J Gastroenterol. 2018;24: 1708-1724. https://doi.org/10.3748/wjg.v24.i16.1708
Lusida MI, Juniastuti, Yano Y. Current hepatitis B virus infection situation in Indonesia and its genetic diversity. World J Gastroenterol. 2016;22: 7264-7274. https://doi.org/10.3748/wjg.v22.i32.7264
Qian F, Qin J, Li D, Ma Z, Zhang H, Jin F, et al. Monitoring of genotypic resistance profile in chronic hepatitis B patients receiving nucleos(t)ide analogues in Huzhou, China. J Infect Dev Ctries. 2016;10: 996-1002. https://doi.org/10.3855/jidc.8020
Tana MM, Ghany MG. Hepatitis B virus treatment: Management of antiviral drug resistance. Clin Liver Dis (Hoboken). 2013;2: 24-28. https://doi.org/10.1002/cld.162
Fu Y, Wu S, Hu Y, Chen T, Zeng Y, Liu C, et al. Mutational characterization of HBV reverse transcriptase gene and the genotype-phenotype correlation of antiviral resistance among Chinese chronic hepatitis B patients. Emerg Microbes Infect. 2020;9: 2381-2393. https://doi.org/10.1080/22221751.2020.1835446
Coppola N, Tonziello G, Colombatto P, Pisaturo M, Messina V, Moriconi F, et al. Lamivudine-resistant HBV strain rtM204V/I in acute hepatitis B. J Infect. 2013;67: 322-328. https://doi.org/10.1016/j.jinf.2013.06.006
Zhang Q, Liao Y, Chen J, Cai B, Su Z, Ying B, et al. Epidemiology study of HBV genotypes and antiviral drug resistance in multi-ethnic regions from Western China. Sci Rep. 2015;5: 17413. https://doi.org/10.1038/srep17413
Araujo NM, Teles SA, Spitz N. Comprehensive analysis of clinically significant hepatitis B virus mutations in relation to genotype, subgenotype and geographic region. Front Microbiol. 2020;11: 616023. https://doi.org/10.3389/fmicb.2020.616023
Long Q-X, Hu J-L, Huang A-L. Deep sequencing of the hepatitis B virus genome: analysis of multiple samples by implementation of the illumina platform. Methods Mol Biol. 2017;1540: 211-218. https://doi.org/10.1007/978-1-4939-6700-1_17
Wang S, Li H, Kou Z, Ren F, Jin Y, Yang L, et al. Highly sensitive and specific detection of hepatitis B virus DNA and drug resistance mutations utilizing the PCR-based CRISPR-Cas13a system. Clin Microbiol Infect. 2021;27: 443-450. https://doi.org/10.1016/j.cmi.2020.04.018
Zhang X, Chen X, Wei M, Zhang C, Xu T, Liu L, et al. Potential resistant mutations within HBV reverse transcriptase sequences in nucleos(t)ide analogues-experienced patients with hepatitis B virus infection. Sci Rep. 2019;9: 8078. https://doi.org/10.1038/s41598-019-44604-6
Fan J, Zhang Y, Xiong H, Wang Y, Guo X. Nucleotide analogue-resistant mutations in hepatitis B viral genomes found in hepatitis B patients. J Gen Virol. 2015;96: 663-670. https://doi.org/10.1099/jgv.0.000010
Arfianti, Zainal A, Andrini F, Endriani R. Distribusi Genotipe dan Subtipe Virus Hepatitis B pada Penderita Hepatitis B Kronik di Pekanbaru. MKB. 2011;43: 105-111. https://doi.org/10.15395/mkb.v43n3.54
Mulyanto, Depamede SN, Surayah K, Tsuda F, Ichiyama K, Takahashi M, et al. A nationwide molecular epidemiological study on hepatitis B virus in Indonesia: identification of two novel subgenotypes, B8 and C7. Arch Virol. 2009;154: 1047-1059. https://doi.org/10.1007/s00705-009-0406-9
Wang L, Han F, Duan H, Ji F, Yan X, Fan Y, et al. Hepatitis B virus pre-existing drug resistant mutation is related to the genotype and disease progression. J Infect Dev Ctries. 2017;11: 727-732. https://doi.org/10.3855/jidc.9021
Copyright (c) 2022 Authors
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
