Objective To elucidate the pathophysiological mechanisms of idiopathic inflammatory myopathy subtypes by analyzing the gene expression profiles of peripheral blood mononuclear cells(PBMCs)from anti-MDA5 antibody-positive and anti-Jo-1 antibody-positive myositis patients.Methods Gene expression profiling screening and analysis of PBMCs from 12 anti-MDA5 positive,16 anti-Jo-1 positive myositis patients and 43 healthy controls were performed using Illumina HT-12 v4 expression profiling microarrays.Applying the unpaired t test with Benjamini-Hochberg correction,the genes with the absolute value of fold change(FC)in gene expression signal ≥2 and adjusted P＜0.05 were selected as differentially expressed genes.Differential gene sets were subjected to Gene Ontology(GO)functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis,with P＜0.05 as the threshold for being significantly enriched.Validation of differentially expressed genes by real time-PCR.The Kolmogorov-Smimov test was used to test the normality of continuous variables.If the distribution was normal and the variance was homogeneous,analysis of variance(one-way ANOVA)was used.If the distribution was not normal,Kruskal-Wallis test was used,and P＜0.05 was regarded as statistically significant difference.Results Analysis of gene expression profiles of PBMCs from patients with positive anti-MDA5 and anti-Jo-1 antibody revealed significant differences in gene expression of PBMCs from patients with the two myositis subtypes.The number of differentially expressed genes that specifically up-regulated in anti-MDA5 antibody positive patients was 407,and the GO functional enrichment analysis was mainly enriched in biological processes such as innate immune response(P＜0.001),response to virus(P＜0.001)and type Ⅰ interferon signaling pathway(P＜0.001),and the KEGG pathway enrichment analysis was mainly enriched in the viral infection-associated pathway(P＜0.001),RIG-Ⅰ like receptor signaling pathway(P＜0.001)and Toll-like receptor signaling pathway(P=0.002),etc.The 259 differential genes specifically down-regulated in the anti-MDA5 antibody positive group were mainly enriched in biological processes such as immune response(P=0.006),TGF-β receptor signaling pathway(P=0.010)and natural killer cell mediated immunity(P=0.015)in GO functional enrichment analysis.There were 162 differentially expressed genes up-regulated specifically in anti-Jo-1 antibody positive patients,and GO functional enrichment analysis was mainly enriched in biological processes such as nucleosome assembly(P＜0.001),negative regulation of cell growth(P=0.001),negative regulation of apoptotic process P=0.004),and innate immune response in mucosa(P=0.012),and the KEGG pathway enrichment analysis mainly enriched in metabolic-related signaling pathways(P＜0.001)and immune-related pathways(P＜0.001),etc.Real-time PCR confirmed that IFIH1(P=0.037),ISG15(P=0.003),and DDX58(P=0.032)in the RIG-Ⅰ-like receptor pathway as well as chemokines MCP-1(P=0.003),MCP-2(P＜0.001),and transcription factor BATF2(P=0.002),and inflammatory signaling pathway-associated MYD88(P＜0.001)were highly expressed in PBMCs from anti-MDA5 antibody-positive myositis patients.Conclusion The gene expression profile of PBMCs in anti-MDA5 antibody-positive patients suggests that the pathogenesis of patients with anti-MDA 5 antibody positive is closely related to biological processes such as innate immune response,viral infection,and interferon response.
MyositisAutoantibodiesGene expression profilingMicrochip analytical proceduresInterferon-stimulated genesRIG-Ⅰ like receptors
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