Aims

Viral integration profiles attract increased interest in the study of HBV-related hepatocellular carcinoma (HCC), but their features in the early stage of infection and changes due to antiviral treatments remain largely unknown.

Methods

Liver biopsies and paired blood samples were obtained from HBeAg-positive patients before and after 48 weeks of entecavir treatment, and a probe-based capture strategy was applied for analyzing the HBV integrations in these samples. Serum HBV markers, including viral DNA, pgRNA, and HBsAg, were longitudinally assessed.

Results

Entecavir treatment successfully reduced the levels of ALT, AST, and HBV serological markers (HBeAg, HBV pgRNA, and HBV DNA) in all patients (<40 years old). As expected, HBV integrations contributed to HBsAg production, with the total number of integrations positively correlated with serum HBsAg level (r = 0.47, P = 0.04). Along with repressed HBV replication, the number of viral integrations in liver biopsies decreased by about 1.94-fold after ETV treatment, with viral breakpoints significantly enriched within nt 1600–1900 of the HBV genome. No recurrent events were observed both at baseline and after treatment for the same individual, and only one same integration was found in two patients. Unlike in tumors, integrations in CHB biopsies seemed to have no chromosomal preference. Moreover, CHB integrations demonstrated lower enrichment scores for open active states than tumors, such as DNase, TssA, and ZNF/Rpts, and the scores reduced after ETV treatment. The antiviral therapy led to the disappearance of the enrichment tendency of integrations in both open chromatin and heterochromatin regions.

Conclusion

Reduced HBV replications by the nucleoside analogue may lead to decreased viral integrations in the liver, and those contributing to the HBsAg production may consistently occur. The pattern of HBV integration after ETV treatment is more random and irregular, which may contribute to a reduced risk of liver cancer due to antiviral treatment.

中文翻译：

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慢性乙型肝炎患者的肝活检表明 HBV 整合谱可能使恩替卡韦治疗的终点和效果复杂化

目标

病毒整合谱吸引了人们对 HBV 相关肝细胞癌 (HCC) 研究的兴趣，但它们在感染早期的特征和抗病毒治疗引起的变化在很大程度上仍然未知。

方法

在恩替卡韦治疗 48 周前后从 HBeAg 阳性患者获得肝活检和配对血液样本，并应用基于探针的捕获策略来分析这些样本中的 HBV 整合。纵向评估了包括病毒 DNA、pgRNA 和 HBsAg 在内的血清 HBV 标志物。

结果

恩替卡韦治疗成功降低了所有患者（<40 岁）的 ALT、AST 和 HBV 血清学标志物（HBeAg、HBV pgRNA 和 HBV DNA）水平。正如预期的那样，HBV 整合有助于 HBsAg 的产生，整合的总数与血清 HBsAg 水平呈正相关（r = 0.47，P = 0.04）。随着 HBV 复制受到抑制，ETV 治疗后肝活检中病毒整合的数量减少了约 1.94 倍，病毒断点在 HBV 基因组的 nt 1600-1900 内显着富集。同一个体在基线和治疗后均未观察到复发事件，并且在两名患者中仅发现了一个相同的整合。与肿瘤不同，CHB 活检中的整合似乎没有染色体偏好。而且，CHB 整合显示开放活性状态的富集分数低于肿瘤，例如 DNase、TssA 和 ZNF/Rpts，并且在 ETV 治疗后分数降低。抗病毒治疗导致开放染色质和异染色质区域整合的富集趋势消失。

结论

核苷类似物减少的 HBV 复制可能导致肝脏中的病毒整合减少，而导致 HBsAg 产生的那些可能持续发生。ETV 治疗后 HBV 整合的模式更加随机和不规则，这可能有助于降低抗病毒治疗后患肝癌的风险。