Hui Rex Wan-Hin, Wong Danny Ka-Ho, Lyu Xueying, Mak Lung-Yi, Fung James, Seto Wai-Kay, Ho Daniel Wai-Hung, Yuen Man-Fung
Department of Medicine, School of Clinical Medicine, The University of Hong Kong, Hong Kong Special Administrative Region of China.
State Key Laboratory of Liver Research, The University of Hong Kong, Hong Kong Special Administrative Region of China.
JHEP Rep. 2025 Jun 17;7(9):101487. doi: 10.1016/j.jhepr.2025.101487. eCollection 2025 Sep.
BACKGROUND & AIMS: HBV integration profiles in the natural history of chronic HBV infection (CHB) have not been well-defined. Hence, we aimed to determine HBV integration profiles across different CHB phases.
We delineated integration profiles from liver biopsies of 55 patients in different CHB phases (3 HBsAg-positive/HBeAg-positive infection; 13 HBsAg-positive/HBeAg-positive hepatitis; 7 HBsAg-positive/HBeAg-negative infection; 12 HBsAg-positive/HBeAg-negative hepatitis; 10 HBsAg seroclearance; 10 occult HBV). Target-capture next-generation sequencing (NovaSeq-6000) was performed, and integrations were characterized on AVID (integrations defined as chimeric fusions in ≥1 soft-clipped reads and ≥2 total reads).
HBV integrations were detected in 35 HBsAg-positive (100%), 8 (80%) HBsAg seroclearance, and 7 (70%) occult HBV patients, respectively. There was a stepwise decrease in integration events from HBsAg-positive/HBeAg-positive (median 9.6 [IQR 9.3-10.1] log integrations per liver), HBsAg-positive/HBeAg-negative (8.7 [8.4-9.0] log integrations) and HBsAg-negative patient groups (7.3 [6.8-7.7] log integrations) (0.001 for trend). There were no differences in integration frequencies in chronic infection (ALT < the upper limit of normal) and chronic hepatitis (ALT ≥ the upper limit of normal) for either HBeAg-positive or -negative patients (all >0.05). No significant differences in integration frequencies were noted between HBsAg seroclearance and occult HBV groups ( >0.05). HBV genome integration breakpoints clustered around nucleotide 1800 in all disease phases. Human genome breakpoints were also delineated, and LINC00486 was the most frequently involved human gene in all disease phases.
We characterized the HBV DNA integration patterns and genome breakpoints of patients in different CHB disease phases. These findings enhance our understanding of the natural history of CHB.
HBV integration profiles in the natural history of chronic HBV infection have not been well-defined. We utilized next-generation sequencing in a well-characterized cohort of patients with HBV at different disease phases, demonstrating a stepwise decrease in integration events as HBV progressed from HBeAg-positive to HBeAg-negative, and then to HBsAg-negative phases. Human and viral genome breakpoints were also identified. These findings enhance our understanding of the natural history of CHB, and provide insights for antiviral treatment.
慢性乙型肝炎病毒（HBV）感染自然史中的HBV整合图谱尚未明确界定。因此，我们旨在确定不同慢性HBV感染阶段的HBV整合图谱。
我们描绘了55例处于不同慢性HBV感染阶段患者的肝脏活检标本中的整合图谱（3例HBsAg阳性/HBeAg阳性感染；13例HBsAg阳性/HBeAg阳性肝炎；7例HBsAg阳性/HBeAg阴性感染；12例HBsAg阳性/HBeAg阴性肝炎；10例HBsAg血清学清除；10例隐匿性HBV感染）。进行了靶向捕获二代测序（NovaSeq - 6000），并在AVID上对整合情况进行了特征分析（整合定义为在≥1条软剪切读段和≥2条总读段中出现嵌合融合）。
分别在35例HBsAg阳性患者（100%）、8例HBsAg血清学清除患者（80%）和7例隐匿性HBV感染患者（70%）中检测到HBV整合。从HBsAg阳性/HBeAg阳性（每肝脏中位数9.6[四分位间距9.3 - 10.1]对数整合）、HBsAg阳性/HBeAg阴性（8.7[8.4 - 9.0]对数整合）和HBsAg阴性患者组（7.3[6.8 - 7.7]对数整合），整合事件呈逐步下降趋势（趋势P = 0.001）。对于HBeAg阳性或阴性患者，慢性感染（ALT <正常上限）和慢性肝炎（ALT≥正常上限）的整合频率无差异（均P>0.05）。HBsAg血清学清除组和隐匿性HBV感染组之间的整合频率无显著差异（P>0.05）。在所有疾病阶段，HBV基因组整合断点聚集在核苷酸1800附近。还描绘了人类基因组断点，LINC00486是所有疾病阶段中最常涉及的人类基因。
我们对不同慢性HBV感染疾病阶段患者的HBV DNA整合模式和基因组断点进行了特征分析。这些发现增进了我们对慢性HBV感染自然史的理解。
慢性HBV感染自然史中的HBV整合图谱尚未明确界定。我们在一组特征明确的不同疾病阶段HBV患者队列中利用二代测序，证明随着HBV从HBeAg阳性进展到HBeAg阴性，再到HBsAg阴性阶段，整合事件逐步减少。还鉴定了人类和病毒基因组断点。这些发现增进了我们对慢性HBV感染自然史的理解，并为抗病毒治疗提供了见解。
