Hepatitis B virus (HBV)-related liver disease induces liver damage by hepatic immune and inflammatory response. The association between aberrant peripheral blood mononuclear cell (PBMC) DNA methylation and progression of liver disease and fibrosis remains unclear. Here we applied Infinium 450 K BeadChip investigating PBMC genome-wide methylation profiling of 48 HBV-related liver disease patients including 24 chronic hepatitis B (CHB), 14 compensated liver cirrhosis (LC), and 10 decompensated liver cirrhosis (DLC). In total, there were 7888 differentially methylated CpG sites (36.06% hypermethylation, 63.94% hypomethylation) correlate with liver disease progression. LC was difficult to be diagnosed, intermediating between CHB and DLC. We used least absolute shrinkage and selection operator (LASSO)-logistic regression method to perform a LC predictive model. The predicted probability (P) of having LC was estimated by the combined model: P = 1/(1 − e−x), where X = 11.52 − 2.82 × (if AST within the normal range − 0.19 × (percent methylation of cg05650055) − 0.21 × (percent methylation of cg17149911 ). Pyrosequencing validation and confusion matrix analysis was used for internal testing, area under receiver operating characteristic curve (AUROC) of model was 0.917 (95% CI, 0.80–0.977). On the fibrosis progress, there were 1705 genes in LC compared with CHB, whose differentially methylated CpG sites loading within the “promoter” regions (including TSS1500, TSS200, 5′UTR, and the 1st exon of genes) subject into the enrichment analysis using Ingenuity Pathway Analysis (IPA). There were 113 enriched immune-related pathways indicated that HBV-related liver fibrosis progression caused epigenetic reprogramming of the immune and inflammatory response. These data support idea that development of HBV-related chronic liver disease is linked with robust and broad alteration of methylation in peripheral immune system. CpG methylation sites serve as relevant biomarker candidates to monitor and diagnose LC, providing new insight into the immune mechanisms understanding the progression of HBV-related liver fibrosis and cirrhosis.

中文翻译：

---

HBV相关肝病在外周血单核细胞DNA甲基化中的特征。

乙型肝炎病毒 (HBV) 相关的肝病通过肝脏免疫和炎症反应诱导肝损伤。异常的外周血单核细胞 (PBMC) DNA 甲基化与肝病和纤维化进展之间的关联仍不清楚。在这里，我们应用 Infinium 450 K BeadChip 研究了 48 名 HBV 相关肝病患者的 PBMC 全基因组甲基化分析，包括 24 名慢性乙型肝炎 (CHB)、14 名代偿性肝硬化 (LC) 和 10 名失代偿性肝硬化 (DLC)。总共有 7888 个差异甲基化的 CpG 位点（36.06% 高甲基化，63.94% 低甲基化）与肝病进展相关。LC 难以诊断，介于 CHB 和 DLC 之间。我们使用最小绝对收缩和选择算子 (LASSO)-逻辑回归方法来执行 LC 预测模型。患有 LC 的预测概率 (P) 由组合模型估计：P = 1/(1 − e-x)，其中 X = 11.52 − 2.82 ×（如果 AST 在正常范围内 − 0.19 ×（cg05650055 的甲基化百分比） ) − 0.21 × (cg17149911 的甲基化百分比)。焦磷酸测序验证和混淆矩阵分析用于内部测试，模型的受试者工作特征曲线下面积 (AUROC) 为 0.917 (95% CI, 0.80–0.977)。关于纤维化进展，与 CHB 相比，LC 中有 1705 个基因，其差异甲基化的 CpG 位点加载在“启动子”区域（包括 TSS1500、TSS200、5'UTR 和基因的第一个外显子）内，使用 Ingenuity Pathway Analysis 进行富集分析（国际音标）。有 113 条丰富的免疫相关通路表明 HBV 相关肝纤维化进展导致免疫和炎症反应的表观遗传重编程。这些数据支持HBV相关慢性肝病的发展与外周免疫系统甲基化的强烈和广泛改变有关的观点。CpG 甲基化位点作为相关生物标志物候选物来监测和诊断 LC，为了解 HBV 相关肝纤维化和肝硬化进展的免疫机制提供了新的见解。这些数据支持HBV相关慢性肝病的发展与外周免疫系统甲基化的强烈和广泛改变有关的观点。CpG 甲基化位点作为相关生物标志物候选物来监测和诊断 LC，为了解 HBV 相关肝纤维化和肝硬化进展的免疫机制提供了新的见解。这些数据支持HBV相关慢性肝病的发展与外周免疫系统甲基化的强烈和广泛改变有关的观点。CpG 甲基化位点作为相关生物标志物候选物来监测和诊断 LC，为了解 HBV 相关肝纤维化和肝硬化进展的免疫机制提供了新的见解。