Chuaypen Natthaya, Payungporn Sunchai, Poovorawan Kittiyod, Chotiyaputta Watcharasak, Piratvisuth Teerha, Tangkijvanich Pisit
Faculty of Medicine, Center of Excellence in Hepatitis and Liver Cancer, Chulalongkorn University, Bangkok, Thailand.
Department of Clinical Tropical Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand.
Virus Genes. 2019 Oct;55(5):610-618. doi: 10.1007/s11262-019-01689-5. Epub 2019 Jul 29.
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.
通过下一代测序（NGS）鉴定的与治疗结果相关的乙型肝炎病毒（HBV）变异体的现有数据有限。本研究旨在确定聚乙二醇干扰素（PEG-IFN）治疗的患者中，HBV C基因型增强子II（EnhII）、核心启动子（BCP）和前核心（PC）区域基线序列变异的作用。纳入接受48周PEG-IFN治疗的HBeAg阳性慢性乙型肝炎（CHB）患者。第96周的联合应答（CR）定义为HBeAg血清学转换加HBV DNA＜2000 IU/mL且HBsAg＜1000 IU/mL。治疗前的病毒突变通过Sanger测序和NGS（Illumina Miseq平台）进行表征。在47例患者（32例男性，平均年龄32.4岁）中，12例（25.5%）实现了CR。总体而言，NGS在检测突变方面优于Sanger测序（61.7%对38.3%，P＜0.001）。基于NGS，与无应答者相比，应答者中T1753V（T1753C/A/G）和A1762T/G1764A变异体的发生率显著更低（分别为8.3%对51.4%，P = 0.009和33.3%对68.6%，P = 0.032）。C1653T和G1896A突变体在两组之间未发现显著差异。不存在T1753V和A1762T/G1764A突变是与CR相关的因素（OR分别为11.65，95%CI 1.36 - 100.16，P = 0.025，以及OR 4.36，9�%CI 1.08 - 17.63，P = 0.039）。治疗前T1753V、A1762T/G1764A突变及其组合的存在分别产生了94.7%、85.7%和93.8%的阴性预测值。基线时通过NGS确定的BCP区域中HBV突变体的存在与接受PEG-IFN的HBeAg阳性CHB患者的治疗效果不佳相关。
