Liang Wenhua, Liu Xin, Chen Zhuxing, Wang Haixuan, Yu Ziwen, Li Chunyan, Yang Hao, Tao Jinsheng, Li Hui, Chen Zhiwei, Fan Jian-Bing, He Jianxing
Department of Thoracic Surgery and Oncology, The First Affiliated Hospital of Guangzhou Medical University, China National Center for Respiratory Medicine, China State Key Laboratory of Respiratory Disease & National Clinical Research Center for Respiratory Disease, Guangzhou, China.
Department of Thoracic Surgery, The First People's Hospital of Foshan, Foshan, China.
J Thorac Dis. 2024 Nov 30;16(11):7592-7606. doi: 10.21037/jtd-24-476. Epub 2024 Nov 13.
Short-term and long-term adverse events could occur after general anesthesia (GA) and the specific mechanism driving these effects has not yet been well-characterized. In this study, we aimed to evaluate the global effect of GA on DNA methylation in the cell-free DNA (cfDNA) of surgical lung-nodule patients.
This large retrospective cohort study enrolled 1,006 surgical lung nodule patients (529 pre-anesthesia, and 477 post-anesthesia). Methylation profiles of the cfDNA isolated from plasma were analyzed by targeted bisulfite sequencing using an enrichment panel covering 12,899 biologically informative methylation regions and 105,844 CpG sites.
By comparing the pre-anesthesia to the post-anesthesia group, a total of 4,562 differentially methylated regions (DMRs) were identified as GA-induced DMRs. Pathway enrichment analysis annotated with cellular processes including pattern specification process, head/heart/bone/tissues development and morphogenesis pathways, cell-adhesion, extra-cellular matrix (ECM) remodeling pathways, and signaling pathways including PI3K-AKT pathway, Ca dependent pathway and RAS/extracellular signal-regulated kinase (RAS/ERK) signaling pathway. Prediction models using 20 DMR markers were derived using Random Forest, which could accurately predict biochemical indicators for post-operative abnormal coagulation function including activated-partial-thromboplastin-time [APTT, area under curve (AUC) 0.81], international normalized ratio (INR, AUC 0.87), D-dimer (AUC 0.82), neutrophil (AUC 0.84) and monocyte (AUC 0.79). Low methylation level in one of the top DMR markers, cg02032606 ( gene), was found to be associated with worse overall survival in both lung adenocarcinoma and squamous carcinoma patients.
This study demonstrated that GA could result in acute DNA methylation changes, which were associated with tissue damage and repair responses. These GA-induced methylation changes were associated with postoperative coagulation functions and could serve as a promising predictive biomarker for coagulation disorders after surgery.
全身麻醉（GA）后可能会发生短期和长期不良事件，而导致这些影响的具体机制尚未得到充分阐明。在本研究中，我们旨在评估GA对外科肺结节患者游离DNA（cfDNA）中DNA甲基化的整体影响。
这项大型回顾性队列研究纳入了1006例外科肺结节患者（麻醉前529例，麻醉后477例）。使用覆盖12899个具有生物学意义的甲基化区域和105844个CpG位点的富集面板，通过靶向亚硫酸氢盐测序分析从血浆中分离的cfDNA的甲基化谱。
通过比较麻醉前组和麻醉后组，共鉴定出4562个差异甲基化区域（DMR）作为GA诱导的DMR。通路富集分析注释了包括模式规范过程、头/心/骨/组织发育和形态发生通路、细胞粘附、细胞外基质（ECM）重塑通路等细胞过程，以及包括PI3K-AKT通路、钙依赖性通路和RAS/细胞外信号调节激酶（RAS/ERK）信号通路等信号通路。使用随机森林得出了使用20个DMR标记的预测模型，该模型可以准确预测术后凝血功能异常的生化指标，包括活化部分凝血活酶时间[APTT，曲线下面积（AUC）0.81]、国际标准化比值（INR，AUC 0.87）、D-二聚体（AUC 0.82）、中性粒细胞（AUC 0.84）和单核细胞（AUC 0.79）。在顶级DMR标记之一cg02032606（基因）中，低甲基化水平被发现与肺腺癌和鳞状细胞癌患者的总体生存率较差有关。
本研究表明，GA可导致急性DNA甲基化变化，这与组织损伤和修复反应有关。这些GA诱导的甲基化变化与术后凝血功能有关，可作为术后凝血障碍的有前景的预测生物标志物。
