Table 3_Multi-omics profiling identifies M1 macrophage polarization-associated biomarkers in hepatitis B virus-related acute-on-chronic liver failure.xlsx

Background and aimsM1 macrophage polarization is essential for the progression of hepatitis B virus-related acute-on-chronic liver failure (HBV-ACLF). In this study, we aim to identify and validate M1 polarization-associated biomarkers to elucidate pathogenic mechanisms and identify potential therapeutic targets for HBV-ACLF.
MethodsMulti-omics data from public databases were analyzed using a co-expression network and two differential expression analyses. Biomarkers were identified by machine learning, ROC curves, and experimental validation. A nomogram was developed to assess the diagnostic efficacy of the biomarkers. Subsequent analyses included functional enrichment, regulatory network construction, computational drug prediction, and molecular docking analysis. Finally, biomarker expression was validated using reverse transcription-quantitative PCR (RT-qPCR) in clinical specimens.
ResultsCDC20, CXCL14, FCGR2B, HKDC1, and GPBAR1 were identified as diagnostic biomarkers for HBV-ACLF. The constructed nomogram showed strong diagnostic performance. Functional enrichment analysis revealed multiple pathways enriched in these biomarkers, including tryptophan metabolism and cofactor biosynthesis, etc. Subsequently, a lncRNA-miRNA-mRNA regulatory network was constructed, with key interactions such as XIST/hsa-miR-296-3p/CXCL14 and SNHG14/hsa-miR-510-5p/CXCL14. Further analysis identified multiple drugs associated with the biomarkers, including cholic acid, deoxycholic acid (GPBAR1-targeting agents). Molecular docking revealed favorable binding affinities between the predicted drugs and their targets, for example, cholic acid exhibited a binding free energy of −7.5 kcal/mol with GPBAR1. In validation experiments, RT-qPCR confirmed significant upregulation of all five biomarkers in HBV-ACLF patients compared with healthy controls (HCs).
ConclusionThis study identifies CDC20, CXCL14, FCGR2B, HKDC1, and GPBAR1 as M1 polarization-associated biomarkers, revealing their roles in immune-metabolic dysregulation and proposing novel therapeutic strategies for HBV-ACLF.

研究背景与目的
M1巨噬细胞极化（M1 macrophage polarization）在乙型肝炎病毒相关慢加急性肝衰竭（HBV-ACLF）的进展中发挥关键作用。本研究旨在鉴定并验证与M1极化相关的生物标志物，以阐明HBV-ACLF的致病机制，并为其寻找潜在治疗靶点。

研究方法
本研究通过共表达网络分析与两种差异表达分析方法，对公共数据库中的多组学数据进行解析。借助机器学习、受试者工作特征曲线（Receiver Operating Characteristic curve, ROC）及实验验证方法筛选生物标志物，构建列线图以评估上述生物标志物的诊断效能。后续分析涵盖功能富集分析、调控网络构建、计算机药物预测及分子对接分析。最终，通过逆转录定量聚合酶链反应（reverse transcription-quantitative PCR, RT-qPCR）对临床标本中的生物标志物表达水平进行验证。

研究结果
本研究鉴定出CDC20、CXCL14、FCGR2B、HKDC1及GPBAR1为HBV-ACLF的诊断生物标志物。所构建的列线图展现出优异的诊断效能。功能富集分析显示，这些生物标志物显著富集于多条通路，包括色氨酸代谢、辅因子生物合成等。随后构建了长链非编码RNA（long non-coding RNA, lncRNA）-微小RNA（microRNA, miRNA）-信使RNA（mRNA）调控网络，其中关键相互作用包括XIST/hsa-miR-296-3p/CXCL14及SNHG14/hsa-miR-510-5p/CXCL14。进一步分析筛选出与这些生物标志物相关的多种药物，包括胆酸、脱氧胆酸（GPBAR1靶向制剂）。分子对接结果显示，预测药物与其靶点之间具有良好的结合亲和力，例如胆酸与GPBAR1的结合自由能为-7.5千卡每摩尔。验证实验中，RT-qPCR证实，与健康对照（healthy controls, HCs）相比，HBV-ACLF患者体内全部5种生物标志物的表达水平均显著上调。

研究结论
本研究鉴定出CDC20、CXCL14、FCGR2B、HKDC1及GPBAR1为与M1巨噬细胞极化相关的生物标志物，揭示了其在免疫代谢紊乱中的作用，并为HBV-ACLF提出了全新的治疗策略。