Liu Sijing, Li Kai, Long Changhai, Lao Mingwu, Ma Biao, Liu Changquan, He Haoyuan, Wang Chunjiang, Chen Wangzhu, Yu Bin
Division of Orthopaedics and Traumatology, Department of Orthopaedics, Nanfang Hospital, Southern Medical University, Guangzhou, China.
Department of Orthopaedic Center, The Second Hospital Affiliated of Guangdong Medical University, Zhanjiang, China.
Front Microbiol. 2025 Feb 6;16:1526475. doi: 10.3389/fmicb.2025.1526475. eCollection 2025.
Regulators of n6-methyladenosine (m6A) RNA modification play important roles in many diseases; however, their involvement in ()-related osteomyelitis remains inadequately explored. Therefore, this study aims to investigate the role of m6A in infection-related osteomyelitis and elucidate its underlying mechanisms.
We downloaded the infection-related osteomyelitis infection dataset GSE30119 from the Gene Expression Omnibus database. Initially, we constructed a diagnostic model based on m6A genes and predicted the hub node miRNAs and transcription factors by constructing a protein-protein interaction network. Subsequently, a prognostic model was built using LASSO regression, the receiver operating characteristic curve of the model was plotted, and the predictive performance of the diagnostic model was validated. Further, unsupervised clustering analysis, gene set enrichment analysis (GSEA), and gene set variation analysis (GSVA) were employed to assess immune cell infiltration. Additionally, we validated the expression of fat mass and obesity-associated protein (FTO) in -infected Raw264.7 macrophages using qPCR and western blotting. Moreover, we conducted si-FTO experiments on mouse Raw264.7 macrophages to investigate the anti-inflammatory regulatory role of si-FTO during infection.
We identified 19 co-expressed genes closely related to FTO were identified, along with 206 related transcription factor regulatory genes and 589 miRNAs. Enrichment analyses suggested that these genes were involved in pathways related to the proliferation and oxidation of various immune cells, cellular senescence, and various tumors and immune cells, as well as cell cycle-related functions. GSEA revealed that PD-1, TH1, TH2, CTLA4, and other pathways were significantly enriched in patients with high FTO expression. GSVA indicated that the differentially enriched pathways were related to included amino acid metabolism, immunity, and infection. Correlation analysis of immune infiltration revealed that monocytes, M2 macrophages, resting mast cells, and neutrophils were present in normal and diseased samples. Differences in expression were observed between the groups. The western blotting and qPCR analyses confirmed that the protein expression of FTO was reduced in macrophages after infection with , consistent with the observed changes in mRNA expression. Furthermore, we validated that FTO may influence the regulation of inflammation through the FoxO1/NF-kB pathway.
The m6A RNA methylation regulator FTO may serve as a potential diagnostic marker and therapeutic target, involved in the pathogenesis of infection-related osteomyelitis. This finding provides new insights into the relationship between FTO-mediated m6A RNA methylation and osteomyelitis.
N6-甲基腺苷（m6A）RNA修饰调节因子在多种疾病中发挥重要作用；然而，它们在（）相关骨髓炎中的作用仍未得到充分研究。因此，本研究旨在探讨m6A在感染相关骨髓炎中的作用，并阐明其潜在机制。
我们从基因表达综合数据库下载了感染相关骨髓炎感染数据集GSE30119。最初，我们基于m6A基因构建了一个诊断模型，并通过构建蛋白质-蛋白质相互作用网络预测了枢纽节点miRNA和转录因子。随后，使用LASSO回归建立了一个预后模型，绘制了该模型的受试者工作特征曲线，并验证了诊断模型的预测性能。此外，采用无监督聚类分析、基因集富集分析（GSEA）和基因集变异分析（GSVA）来评估免疫细胞浸润。另外，我们使用qPCR和蛋白质免疫印迹法验证了脂肪量和肥胖相关蛋白（FTO）在感染的Raw264.7巨噬细胞中的表达。此外，我们对小鼠Raw264.7巨噬细胞进行了si-FTO实验，以研究si-FTO在感染期间的抗炎调节作用。
我们鉴定出19个与FTO密切相关的共表达基因，以及206个相关转录因子调节基因和589个miRNA。富集分析表明，这些基因参与了与各种免疫细胞的增殖和氧化、细胞衰老、各种肿瘤和免疫细胞以及细胞周期相关功能有关的途径。GSEA显示，在FTO高表达患者中，PD-1、TH1、TH2、CTLA4等途径显著富集。GSVA表明，差异富集途径与氨基酸代谢、免疫和感染有关。免疫浸润的相关性分析显示，正常和患病样本中存在单核细胞、M2巨噬细胞、静息肥大细胞和中性粒细胞。两组之间观察到表达差异。蛋白质免疫印迹法和qPCR分析证实，感染后巨噬细胞中FTO的蛋白表达降低，这与观察到的mRNA表达变化一致。此外，我们验证了FTO可能通过FoxO1/NF-κB途径影响炎症调节。
m6A RNA甲基化调节因子FTO可能作为一种潜在的诊断标志物和治疗靶点，参与感染相关骨髓炎的发病机制。这一发现为FTO介导的m6A RNA甲基化与骨髓炎之间的关系提供了新的见解。
