基于抗体的特发性炎性肌病患者外周血基因表达谱分析  

Antibody-based gene expression profiling of peripheral blood from patients with idiopathic inflammatory myopathy

作　　者：李牧原[1] 王莉 李全贞 罗卉[1] 张华莉 Li Muyuan;Wang Li;Li Quanzhen;Luo Hui;Zhang Huali(Department of Rheumatology,Xiangya Hospital,Central South University,Changsha 410008,China;Department of Pathophysiology,Xiangya School of Medicine,Central South University,Changsha 410013,China)

机构地区：[1]中南大学湘雅医院风湿免疫科,长沙410008 [2]中南大学基础医学院病理生理学系,长沙410013

出　　处：《中华风湿病学杂志》2024年第1期15-21,I0004-I0006,共10页Chinese Journal of Rheumatology

基　　金：国家自然科学基金(82070018)。

摘　　要：目的通过对抗黑色素瘤分化相关基因5(MDA5)抗体阳性和抗Jo-1抗体阳性的肌炎患者外周血单个核细胞(PBMCs)进行基因表达谱的分析,以阐明特发性炎性肌病亚型的病理生理学机制。方法①用Illumina HT-12 v4表达谱芯片对12例抗MDA5抗体阳性和16例抗Jo-1抗体阳性的肌炎患者及43名健康对照者的PBMCs进行基因表达谱筛查和分析。应用非配对t检验,经Benjamini-Hochberg校正,选取基因表达信号倍数变化的绝对值≥2且校正P<0.05为差异表达基因。将差异基因集进行基因本体论数据库(GO)功能富集分析和京都基因和基因组百科全书(KEGG)通路富集分析,以P<0.05作为显著性富集的阈值。②用实时荧光聚合酶链反应(real time-PCR)对差异表达基因进行验证,采用Kolmogorov-Smirnov检验对连续型变量进行正态性检验,符合正态分布且方差齐性,采用单因素方差分析,不符合正态分布则用Kruskal-Wallis检验,P<0.05为差异有统计学意义。结果①分析抗MDA5抗体和抗Jo-1抗体阳性患者PBMCs的基因表达谱,发现2种肌炎亚型患者PBMCs的基因表达存在明显差异。抗MDA5抗体阳性患者特异性表达上调的差异基因为407个,GO功能富集分析主要富集于固有免疫应答(P<0.001)、病毒应答(P<0.001)和干扰素应答(P<0.001)等生物过程,KEGG通路富集分析主要富集于病毒感染相关通路(P<0.001)、视黄酸诱导基因Ⅰ(RIG-Ⅰ)样受体通路(P<0.001)和Toll样受体通路(P=0.002)等。抗MDA5抗体阳性组中特异性下调的259个差异基因,GO功能富集分析主要富集于免疫应答(P=0.006)、TGF-β受体信号通路(P=0.010)和自然杀伤细胞介导的免疫(P=0.015)等生物过程。抗Jo-1抗体阳性患者特异性表达上调的差异基因有162个,GO功能富集分析主要富集于核小体组装(P<0.001)、细胞增长的负调控(P=0.001)、凋亡负调控(P=0.004)和黏膜固有免疫(P=0.012)等生物过程,KEGG通路富集分析主要富集于代谢�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-Smirnov 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 typeⅠinterferon signaling pathway(P<0.001),and the KEGG pathway enrichment analysis was mainly enriched in the viral infection-associated pathway(P<0.001),RIG-Ⅰlike receptor signaling pathway(P<0.001)and Toll-like recept

关 键 词：肌炎 自身抗体 基因表达谱 芯片分析技术 干扰素刺激基因 RIG-Ⅰ样受体 

分 类 号：R593.26[医药卫生—内科学]