{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T02:06:19Z","timestamp":1788919579347,"version":"build-2803163510"},"reference-count":51,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,2,6]],"date-time":"2025-02-06T00:00:00Z","timestamp":1738800000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Microbiol."],"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p>Regulators of n6-methyladenosine (m6A) RNA modification play important roles in many diseases; however, their involvement in <jats:italic>Staphylococcus aureus<\/jats:italic> (<jats:italic>S. aureus<\/jats:italic>)-related osteomyelitis remains inadequately explored. Therefore, this study aims to investigate the role of m6A in <jats:italic>S. aureus<\/jats:italic> infection-related osteomyelitis and elucidate its underlying mechanisms.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>We downloaded the <jats:italic>S. aureus<\/jats:italic> 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\u2013protein 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 <jats:italic>S. aureus<\/jats:italic>-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 <jats:italic>S. aureus<\/jats:italic> infection.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>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 <jats:italic>S. aureus<\/jats:italic>, 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.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>The m6A RNA methylation regulator FTO may serve as a potential diagnostic marker and therapeutic target, involved in the pathogenesis of <jats:italic>S. aureus<\/jats:italic> infection-related osteomyelitis. This finding provides new insights into the relationship between FTO-mediated m6A RNA methylation and osteomyelitis.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fmicb.2025.1526475","type":"journal-article","created":{"date-parts":[[2025,2,6]],"date-time":"2025-02-06T07:11:41Z","timestamp":1738825901000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":11,"title":["The role of FTO in m6A RNA methylation and immune regulation in Staphylococcus aureus infection-related osteomyelitis"],"prefix":"10.3389","volume":"16","author":[{"given":"Sijing","family":"Liu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kai","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Changhai","family":"Long","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mingwu","family":"Lao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Biao","family":"Ma","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Changquan","family":"Liu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haoyuan","family":"He","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chunjiang","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wangzhu","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bin","family":"Yu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,2,6]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"25","DOI":"10.1038\/75556","article-title":"Gene ontology: tool for the unification of biology. The Gene Ontology Consortium","volume":"25","author":"Ashburner","year":"2000","journal-title":"Nat. Genet."},{"key":"ref2","doi-asserted-by":"publisher","first-page":"3234","DOI":"10.3390\/ijms24043234","article-title":"Staphylococcus aureus genomic analysis and outcomes in patients with bone and joint infections: a systematic review","volume":"24","author":"Bouiller","year":"2023","journal-title":"Int. J. Mol. Sci."},{"key":"ref3","doi-asserted-by":"publisher","first-page":"W244","DOI":"10.1093\/nar\/gkaa467","article-title":"miRNet 2.0: network-based visual analytics for miRNA functional analysis and systems biology","volume":"48","author":"Chang","year":"2020","journal-title":"Nucleic Acids Res."},{"key":"ref4","doi-asserted-by":"publisher","first-page":"248","DOI":"10.1016\/j.celrep.2016.12.019","article-title":"Pan-cancer Immunogenomic analyses reveal genotype-Immunophenotype relationships and predictors of response to checkpoint blockade","volume":"18","author":"Charoentong","year":"2017","journal-title":"Cell Rep."},{"key":"ref5","doi-asserted-by":"publisher","first-page":"474","DOI":"10.3389\/fimmu.2019.00474","article-title":"DNA sensor IFI204 contributes to host defense against Staphylococcus aureus infection in mice","volume":"10","author":"Chen","year":"2019","journal-title":"Front. Immunol."},{"key":"ref6","doi-asserted-by":"publisher","first-page":"90","DOI":"10.1136\/gutjnl-2024-331903","article-title":"Targeting the oncogenic m6A demethylase FTO suppresses tumourigenesis and potentiates immune response in hepatocellular carcinoma","volume":"74","author":"Chen","year":"2024","journal-title":"Gut"},{"key":"ref7","doi-asserted-by":"publisher","first-page":"103","DOI":"10.1186\/s12943-019-1033-z","article-title":"The role of m6A RNA methylation in human cancer","volume":"18","author":"Chen","year":"2019","journal-title":"Mol. Cancer"},{"key":"ref8","doi-asserted-by":"publisher","first-page":"11","DOI":"10.1186\/1752-0509-8-S4-S11","article-title":"cytoHubba: identifying hub objects and sub-networks from complex interactome","volume":"8","author":"Chin","year":"2014","journal-title":"BMC Syst. Biol."},{"key":"ref9","doi-asserted-by":"publisher","first-page":"3971","DOI":"10.1073\/pnas.71.10.3971","article-title":"Identification of methylated nucleosides in messenger RNA from Novikoff hepatoma cells","volume":"71","author":"Desrosiers","year":"1974","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref10","doi-asserted-by":"publisher","first-page":"1596","DOI":"10.1007\/s12035-018-1138-1","article-title":"m6A RNA methylation controls neural development and is involved in human diseases","volume":"56","author":"Du","year":"2019","journal-title":"Mol. Neurobiol."},{"key":"ref11","doi-asserted-by":"publisher","first-page":"652","DOI":"10.1186\/s12967-023-04461-4","article-title":"FTO-targeted siRNA delivery by MSC-derived exosomes synergistically alleviates dopaminergic neuronal death in Parkinson\u2019s disease via m6A-dependent regulation of ATM mRNA","volume":"21","author":"Geng","year":"2023","journal-title":"J. Transl. Med."},{"key":"ref12","doi-asserted-by":"publisher","first-page":"1469","DOI":"10.1126\/science.1151710","article-title":"(2017). The obesity-associated FTO gene encodes a 2-oxoglutarate-dependent nucleic acid demethylase","volume":"318","author":"Gerken","year":"2007","journal-title":"Science"},{"key":"ref13","doi-asserted-by":"publisher","first-page":"1408","DOI":"10.1126\/science.aad8711","article-title":"Messenger RNA modifications: form, distribution, and function","volume":"352","author":"Gilbert","year":"2016","journal-title":"Science"},{"key":"ref14","doi-asserted-by":"publisher","first-page":"7","DOI":"10.1186\/1471-2105-14-7","article-title":"GSVA: gene set variation analysis for microarray and RNA-seq data","volume":"14","author":"H\u00e4nzelmann","year":"2023","journal-title":"BMC Bioinformatics"},{"key":"ref15","first-page":"1027","article-title":"Diagnosis and management of osteomyelitis","volume":"84","author":"Hatzenbuehler","year":"2021","journal-title":"Am. Fam. Physician"},{"key":"ref16","doi-asserted-by":"publisher","first-page":"5753","DOI":"10.7150\/ijbs.74196","article-title":"N6-methyladenosine of Socs1 modulates macrophage inflammatory response in different stiffness environments","volume":"18","author":"Hu","year":"2022","journal-title":"Int. J. Biol. Sci."},{"key":"ref17","doi-asserted-by":"publisher","first-page":"94","DOI":"10.1016\/j.nutres.2017.12.005","article-title":"Higher prevalence of FTO gene risk genotypes AA rs9939609, CC rs1421085, and GG rs17817449 and saliva containing Staphylococcus aureus in obese women in Croatia","volume":"50","author":"Hu\u0111ek","year":"2017","journal-title":"Nutr. Res."},{"key":"ref18","doi-asserted-by":"publisher","first-page":"839","DOI":"10.1038\/s41598-020-57619-1","article-title":"Potential osteomyelitis biomarkers identified by plasma metabolome analysis in mice","volume":"10","author":"Isogai","year":"2020","journal-title":"Sci. Rep."},{"key":"ref19","doi-asserted-by":"publisher","first-page":"3313","DOI":"10.1016\/j.febslet.2008.08.019","article-title":"Oxidative demethylation of 3-methylthymine and 3-methyluracil in single-stranded DNA and RNA by mouse and human FTO","volume":"582","author":"Jia","year":"2008","journal-title":"FEBS Lett."},{"key":"ref20","doi-asserted-by":"publisher","first-page":"27","DOI":"10.1093\/nar\/28.1.27","article-title":"KEGG: Kyoto encyclopedia of genes and genomes","volume":"28","author":"Kanehisa","year":"2000","journal-title":"Nucleic Acids Res."},{"key":"ref21","doi-asserted-by":"publisher","first-page":"882","DOI":"10.1093\/bioinformatics\/bts034","article-title":"The sva package for removing batch effects and other unwanted variation in high-throughput experiments","volume":"28","author":"Leek","year":"2012","journal-title":"Bioinformatics"},{"key":"ref22","doi-asserted-by":"publisher","first-page":"167341","DOI":"10.1016\/j.bbadis","article-title":"FTO-mediated RNA m6A methylation regulates synovial aggression and inflammation in rheumatoid arthritis","volume":"1870","author":"Li","year":"2024","journal-title":"Biochim. Biophys. Acta Mol. basis Dis."},{"key":"ref23","doi-asserted-by":"publisher","first-page":"113712","DOI":"10.1016\/j.yexcr.2023.113712","article-title":"m6A-mediated upregulation of miRNA-193a aggravates cardiomyocyte apoptosis and inflammatory response in sepsis-induced cardiomyopathy via the METTL3\/ miRNA-193a\/BCL2L2 pathway","volume":"430","author":"Liang","year":"2023","journal-title":"Exp. Cell Res."},{"key":"ref24","doi-asserted-by":"publisher","first-page":"417","DOI":"10.1016\/j.cels.2015.12.004","article-title":"The molecular signatures database (MSigDB) hallmark gene set collection","volume":"1","author":"Liberzon","year":"2015","journal-title":"Cell Syst."},{"key":"ref25","doi-asserted-by":"publisher","first-page":"1187543","DOI":"10.3389\/fcimb.2023.1187543","article-title":"Apolipoprotein E deficiency potentiates macrophage against Staphylococcus aureus in mice with osteomyelitis via regulating cholesterol metabolism","volume":"13","author":"Lu","year":"2023","journal-title":"Front. Cell. Infect. Microbiol."},{"key":"ref26","doi-asserted-by":"publisher","first-page":"339","DOI":"10.1186\/s12967-023-04173-9","article-title":"Expression pattern analysis of m6A regulators reveals IGF2BP3 as a key modulator in osteoarthritis synovial macrophages","volume":"21","author":"Lu","year":"2023","journal-title":"J. Transl. Med."},{"key":"ref27","doi-asserted-by":"publisher","first-page":"663295","DOI":"10.3389\/fimmu.2021.663295","article-title":"Targeted inhibition of FTO demethylase protects mice against LPS-induced septic shock by suppressing NLRP3 Inflammasome","volume":"12","author":"Luo","year":"2021","journal-title":"Front. Immunol."},{"key":"ref28","doi-asserted-by":"publisher","first-page":"11","DOI":"10.1186\/1471-2121-15-11","article-title":"Inhibition of methicillin-resistant Staphylococcus aureus-induced cytokines mRNA production in human bone marrow derived mesenchymal stem cells by 1,25-dihydroxyvitamin D3","volume":"15","author":"Maiti","year":"2014","journal-title":"BMC Cell Biol."},{"key":"ref29","doi-asserted-by":"publisher","first-page":"29","DOI":"10.1016\/j.jgar.2021.03.030","article-title":"Staphylococcus aureus clones causing osteomyelitis: a literature review","volume":"26","author":"Pimentel de Araujo","year":"2021","journal-title":"J. Glob. Antimicrob. Resist."},{"key":"ref30","doi-asserted-by":"publisher","first-page":"697026","DOI":"10.3389\/fimmu.2021.697026","article-title":"N6-Methyladenosine in Cancer immunotherapy: an undervalued therapeutic target","volume":"12","author":"Quan","year":"2021","journal-title":"Front. Immunol."},{"key":"ref31","doi-asserted-by":"publisher","first-page":"e47","DOI":"10.1093\/nar\/gkv007","article-title":"Limma powers differential expression analyses for RNA-sequencing and microarray studies","volume":"43","author":"Ritchie","year":"2015","journal-title":"Nucleic Acids Res."},{"key":"ref32","doi-asserted-by":"publisher","first-page":"77","DOI":"10.1186\/1471-2105-12-77","article-title":"pROC: an open-source package for R and S+ to analyze and compare ROC curves","volume":"12","author":"Robin","year":"2011","journal-title":"BMC Bioinformatics"},{"key":"ref33","doi-asserted-by":"publisher","first-page":"2498","DOI":"10.1101\/gr.1239303","article-title":"Cytoscape: a software environment for integrated models of biomolecular interaction networks","volume":"13","author":"Shannon","year":"2003","journal-title":"Genome Res."},{"key":"ref34","doi-asserted-by":"publisher","first-page":"1805","DOI":"10.2147\/JIR.S406562","article-title":"Identification of Ferroptosis-related biomarkers for diagnosis and molecular classification of Staphylococcus aureus-induced osteomyelitis","volume":"16","author":"Shi","year":"2023","journal-title":"J. Inflamm. Res."},{"key":"ref35","doi-asserted-by":"publisher","first-page":"640","DOI":"10.1016\/j.molcel.2019.04.025","article-title":"Where, when, and how: context-dependent functions of RNA methylation writers, readers, and erasers","volume":"74","author":"Shi","year":"2019","journal-title":"Mol. Cell"},{"key":"ref36","doi-asserted-by":"publisher","first-page":"135","DOI":"10.1007\/978-1-0716-0301-7_7","article-title":"Profiling cell type abundance and expression in bulk tissues with CIBERSORTx","volume":"2117","author":"Steen","year":"2020","journal-title":"Methods Mol. Biol."},{"key":"ref37","doi-asserted-by":"publisher","first-page":"1.30.1","DOI":"10.1002\/cpbi.5","article-title":"The GeneCards suite: from gene data mining to disease genome sequence analyses","volume":"54","author":"Stelzer","year":"2016","journal-title":"Curr. Protoc. Bioinformatics"},{"key":"ref38","doi-asserted-by":"publisher","first-page":"15545","DOI":"10.1073\/pnas.0506580102","article-title":"Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles","volume":"102","author":"Subramanian","year":"2005","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"key":"ref39","doi-asserted-by":"publisher","first-page":"291","DOI":"10.1111\/imm.13239","article-title":"Annexin A3 in sepsis: novel perspectives from an exploration of public transcriptome data","volume":"161","author":"Toufiq","year":"2000","journal-title":"Immunology"},{"key":"ref40","doi-asserted-by":"publisher","first-page":"258","DOI":"10.1093\/nar\/gkg034","article-title":"STRING: a database of predicted functional associations between proteins","volume":"31","author":"Von Mering","year":"2003","journal-title":"Nucleic Acids Res."},{"key":"ref42","doi-asserted-by":"publisher","first-page":"1572","DOI":"10.1093\/bioinformatics\/btq170","article-title":"ConsensusClusterPlus: a class discovery tool with confidence assessments and item tracking","volume":"26","author":"Wilkerson","year":"2010","journal-title":"Bioinformatics"},{"key":"ref43","first-page":"1957","article-title":"N6-methyladenine RNA modification and cancers","volume":"8","author":"Wu","year":"2018","journal-title":"Am. J. Cancer Res."},{"key":"ref44","doi-asserted-by":"publisher","first-page":"602485","DOI":"10.3389\/fgene.2020.602485","article-title":"The identification of critical m6A RNA methylation regulators as malignant prognosis factors in prostate adenocarcinoma","volume":"11","author":"Xu","year":"2020","journal-title":"Front. Genet."},{"key":"ref45","doi-asserted-by":"publisher","first-page":"960","DOI":"10.1038\/s41419-020-03143-z","article-title":"The role of m6A modification in physiology and disease","volume":"11","author":"Yang","year":"2020","journal-title":"Cell Death Dis."},{"key":"ref46","doi-asserted-by":"publisher","first-page":"2782","DOI":"10.1038\/s41467-019-10669-0","article-title":"m6A mRNA demethylase FTO regulates melanoma tumorigenicity and response to anti-PD-1 blockade","volume":"10","author":"Yang","year":"2019","journal-title":"Nat. Commun."},{"key":"ref47","doi-asserted-by":"publisher","first-page":"D682","DOI":"10.1093\/nar\/gkz966","article-title":"Ensembl 2020","volume":"48","author":"Yates","year":"2000","journal-title":"Nucleic Acids Res."},{"key":"ref48","doi-asserted-by":"publisher","first-page":"284","DOI":"10.1089\/omi.2011.0118","article-title":"clusterProfiler: an R package for comparing biological themes among gene clusters","volume":"16","author":"Yu","year":"2012","journal-title":"OMICS"},{"key":"ref49","doi-asserted-by":"publisher","first-page":"481","DOI":"10.1097\/BPB.0000000000001041","article-title":"(2023). Meta-analysis of serum procalcitonin diagnostic test accuracy for osteomyelitis and septic arthritis in children","volume":"32","author":"Zhang","year":"2023","journal-title":"J. Pediatr. Orthop. B"},{"key":"ref50","doi-asserted-by":"publisher","first-page":"244","DOI":"10.1186\/1471-2105-14-244","article-title":"RCircos: an R package for Circos 2D track plots","volume":"14","author":"Zhang","year":"2013","journal-title":"BMC Bioinformatics"},{"key":"ref51","doi-asserted-by":"publisher","first-page":"4837","DOI":"10.2147\/OTT.S329232","article-title":"Roles of N6-Methyladenosine demethylase FTO in malignant tumors progression","volume":"14","author":"Zheng","year":"2021","journal-title":"Onco. Targets. Ther."},{"key":"ref52","doi-asserted-by":"publisher","first-page":"e160517","DOI":"10.1172\/JCI160517","article-title":"FTO fuels diabetes-induced vascular endothelial dysfunction associated with inflammation by erasing m6A methylation of TNIP1","volume":"133","author":"Zhou","year":"2023","journal-title":"J. Clin. Invest."}],"container-title":["Frontiers in Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmicb.2025.1526475\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,2,6]],"date-time":"2025-02-06T07:11:48Z","timestamp":1738825908000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fmicb.2025.1526475\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,2,6]]},"references-count":51,"alternative-id":["10.3389\/fmicb.2025.1526475"],"URL":"https:\/\/doi.org\/10.3389\/fmicb.2025.1526475","relation":{},"ISSN":["1664-302X"],"issn-type":[{"value":"1664-302X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,2,6]]},"article-number":"1526475"}}