Abstract
Current data of hepatitis B virus (HBV) variants associated with treatment outcome identified by next generation sequencing (NGS) are limited. This study was aimed at determining the role of baseline sequence variations in the enhancer II (EnhII), basal core promotor (BCP) and pre-core (PC) regions of HBV genotype C in patients treated with pegylated interferon (PEG-IFN). Patients with HBeAg-positive chronic hepatitis B (CHB) treated with 48-week PEG-IFN were enrolled. Combined response (CR) at week 96 was defined by HBeAg seroconversion plus HBV DNA < 2000 IU/mL and HBsAg < 1000 IU/mL. Pre-treatment viral mutations were characterized by Sanger sequencing and NGS (Miseq Illumina platform). Among 47 patients (32 male, mean age 32.4 years), CR was achieved in 12 (25.5%) individuals. Overall, NGS was superior to Sanger sequencing in detecting mutations (61.7% vs. 38.3%, P < 0.001). Based on NGS, the prevalence of T1753V (T1753C/A/G) and A1762T/G1764A variants were significantly lower in responders compared to non-responders (8.3% vs. 51.4%, P = 0.009 and 33.3% vs. 68.6%, P = 0.032, respectively). No significant difference between groups was found regarding C1653T and G1896A mutants. The absence of T1753V and A1762T/G1764A mutations were factors associated with CR (OR 11.65, 95%CI 1.36–100.16, P = 0.025, and OR 4.36, 95%CI 1.08–17.63, P = 0.039, respectively). The existence of pre-treatment T1753V, A1762T/G1764A mutations and their combination yielded negative predictive values of 94.7%, 85.7% and 93.8%, respectively. The presence of HBV mutants in the BCP region determined by NGS at baseline was associated with poor treatment outcome in patients with HBeAg-positive CHB receiving PEG-IFN.
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Acknowledgements
This study was funded by the Grant for Chula Research Scholar (CU-GRS-60-06-30-03) and Postdoctoral Fellowship under Rachadapisek Sompot Fund, Chulalongkorn University. The study was also supported by Center of Excellence in Hepatitis and Liver Cancer, Chulalongkorn University, The Thailand Research Fund (RTA5980008) Senior Research Scholar and the Thai Association for the Study of the Liver (THASL).
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NC performed the experiments, analyzed statistical data, wrote the first draft of the manuscript. SP was involved in experimental design and data analysis. KP, WC, and TP collaborated in the clinical data analysis and reviewed draft manuscript. PT developed experimental design, main analysis plan, and edited manuscript prior to submission. All co-author read and approved the final manuscript.
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All subjects had provided written informed consent as approved by the Institutional Review Board of the Faculty of Medicine, Chulalongkorn University (IRB no 016/59), which followed the Helsinki Declaration and Good Clinical Practice guidelines.
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11262_2019_1689_MOESM1_ESM.tif
Supplementary Fig. 1: Hotspot mutations across the EnhII/BCP/PC regions in relation to treatment response. Supplementary material 1 (TIFF 1151 kb)
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Supplementary Fig. 2: Baseline and changes of HBsAg levels during and after treatment regarding to T1753V and A1762T/G1764A variants. Supplementary material 2 (TIFF 600 kb)
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Chuaypen, N., Payungporn, S., Poovorawan, K. et al. Next generation sequencing identifies baseline viral mutants associated with treatment response to pegylated interferon in HBeAg-positive chronic hepatitis B. Virus Genes 55, 610–618 (2019). https://doi.org/10.1007/s11262-019-01689-5
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DOI: https://doi.org/10.1007/s11262-019-01689-5