General anesthesia induces acute cell-free DNA methylation changes in peripheral blood.

BACKGROUND: 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. METHODS: 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. RESULTS: 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(2+) 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 (DLX-4 gene), was found to be associated with worse overall survival in both lung adenocarcinoma and squamous carcinoma patients. CONCLUSIONS: 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.