Objective To explore the potential pathogenesis of exosome-mediated polymyositis/dermatomyositis(PM/DM)through multi-omics combined with bioinformatic analysis approach and to identify potential new targets for the diagnosis and treatment of PM/DM.Methods Collect serum exosome samples from PM/DM patients and healthy individuals who underwent physical examination in Wuxi Second People's Hospital from January 2021 to June 2023.HiSeq high-throughput sequencing technology and iTRAQ quantitative proteomics techniques were used to perform a sequencing analysis of miRNA and protein components in serum exosomes from patients with PM/DM and healthy control.R language was adapted to conduct the limma differential analysis,gene ontology(GO),Kyoto encyclopedia of genes and genomes(KEGG),gene set enrichment analysis(GSEA)functional analysis,and immunological correlation analysis.Based on these analysis,we identified core genes and established a miRNA-target gene-transcription factor co-expression molecular network.Subsequently,we employed quantitative real-time PCR(qRT-PCR)to experimentally validate the core genes.Data analysis was performed using t-test statistical analysis and receiver operating characteristic(ROC)curves were plotted to evaluate the test efficacy of the core genes.Results Initially,42 up-regulated differential proteins and 61 DEP down-regulated differential proteins,as well as 22 up-regulated differential miRNAs and 19 down-regulated miRNAs were screened,and 7 core genes,13 associated differential miRNAs,and 4 transcription factors were finally identified.Based on the functional analysis we concluded that the core genes CTSG,MPO,Hl-5 involved in the formation of neutrophil extracellular trap network,NF-κB pathway and other inflammation-related pathways might play an important role in exosome-mediated PM/DM pathogenesis.Immune cells such as mast cells,immature dendritic cells,natural killer cells,regulatory T cells,and helper T cells 2 were expressed to a higher extent in the disease.In the t-test,MPO[(1.08±0.47)vs.(2.05±0.62),t=-3.50,P=0.004],CTSG[(1.11±0.51)vs.(2.27±1.10),t=-2.72,P=0.022],and H1-5[(1.03±0.25)vs.(1.81±0.73),t=-2.89,P=0.019]showed statistically significant differences.In the ROC curve analysis,MPO[AUC(95％CI):0.92(0.78,1)],CTSG[AUC(95％CI):0.81(0.59,1)],and H1-5(AUC(95％CI)=0.84(0.64,1)]indicated that the three core genes had good accuracy.Conclusion We identified the key differential molecules in serum exosomes of patients with PM/DM,and constructed a regulatory network of miRNA-target gene-transcription factor,and determined that the pathogenic mechanism of PM/DM was mediated by serum exosomes was mediated through the formation of neutrophil extracellular trapping nets and the NF-κB pathway.CTSG,MPO,and Hl-5 are the core genes in the these pathways,and their related miRNAs and transcription factors have been identified,which may become potential targets and biomarkers for the diagnosis and treatment of PM/DM.
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