基于RNA测序的A亚型呼吸道合胞病毒毛细支气管炎转录组学特征分析及关键基因预测  

Transcriptome characteristics analysis and Hub genes prediction of respiratory syncytial virus subtype A bronchiolitis based on RNA sequencing

作　　者：李海燕[1] 翁婷婷 王乐颖 梁春婵 董琳[1] LI Haiyan;WENG Tingting;WANG Leying;LIANG Chunchan;DONG Lin(Department of Pediatric Pulmonology,the Second Affiliated Hospital&Yuying Children’s Hospital of Wenzhou Medical University,Wenzhou 325027,China)

机构地区：[1]温州医科大学附属第二医院育英儿童医院儿童呼吸科,浙江温州325027

出　　处：《温州医科大学学报》2022年第10期784-791,共8页Journal of Wenzhou Medical University

基　　金：温州市基础性科研项目(Y20190437);浙江省公益技术研究计划项目(LGF20H010004);温州医科大学附属第二医院育英儿童医院贺林院士工作站科研基金重点项目(18331103)。

摘　　要：目的:基于RNA测序数据的分析,寻找A亚型呼吸道合胞病毒(RSV)毛细支气管炎有关的关键基因(Hub基因)和通路,为RSV毛细支气管炎的干预和治疗提供依据。方法:招募23例RSV毛细支气管炎住院患儿为病例组,以10例健康儿童为对照组。收集外周血白细胞并提取RNA用于高通量测序。利用三种R软件包(DESeq2、edgeR、limma)获取病例组和对照组之间的差异表达基因(DEGs)。利用R包clusterProfiler和Metascape在线工具对所有的DEGs进行GO和KEGG富集分析。利用STRING数据库进行DEGs的蛋白互作分析,利用Cytoscape软件筛选Hub基因。结果:共筛选出286个DEGs,包括166个表达上调的基因和120个表达下调的基因。GO功能富集分析发现DEGs主要参与过氧化氢代谢过程、细胞对生物刺激的应激反应、维生素D受体信号通路等生物学过程。KEGG通路分析显示DEGs主要涉及免疫系统中的细胞因子信号、适应性免疫系统、中性粒细胞脱颗粒等信号通路。从蛋白互作网络中筛选出4个核心模块,主要与RAF/MAP激酶级联反应、病毒蛋白与细胞因子和细胞因子受体互作等通路有关。最后筛选出RRM2、BUB1B、BUB1、MCM10、CDC45、MKI67、ASPM、NCAPG、NUSAP1、ESPL1、CDT1共11个与A亚型RSV毛细支气管炎相关的Hub基因。结论:本研究鉴定出的免疫相关通路和Hub基因可能与RSV毛细支气管炎发病有关,其具体机制值得进一步研究。Objective:To screen potential key genes(Hub genes)and pathways involved in the pathogenesis of bronchiolitis in children caused by respiratory syncytial virus(RSV)subtype A using RNA sequencing data analysis,aiming to provide scientific clues for intervention and treatment of RSV bronchiolitis.Methods:Twenty-three patients with RSV bronchiolitis(case group)and ten healthy children(healthy control)were enrolled.RNA was extracted from peripheral blood leukocytes for high-throughput RNA sequencing.The differentially expressed genes(DEGs)between case and control were obtained by three R packages including DESeq2,edgeR and limma.All DEGs were selected for GO and KEGG enrichment analysis by“clusterProfiler”package and Metascape tool,respectively.Protein-protein interaction(PPI)analysis for all DEGs was constructed using STRING database,and finally Hub genes were identified in Cytoscape software.Results:We screened a total of 286 DEGs between case and control,including 166 upregulated genes and 120 downregulated genes.GO enrichment analysis showed that these DEGs were mainly enriched in several biological processes including hydrogen peroxide catabolic process,cellular response to biotic stimulus and vitamin D receptor signaling pathway.KEGG pathway analysis showed that these DEGs were mainly involved in several pathways related to immune response,such as cytokine signaling in immune system,adaptive immune system and neutrophil degranulation.Using the MCODE plug-in of Cytoscape,we determined four core modules from entire PPI network,mainly associated with viral protein interaction with cytokine and cytokine receptor,and RAF/MAP kinase cascade.Finally,we screened a total of 11 Hub genes related to RSV subtype A bronchiolitis,including RRM2,BUB1B,BUB1,MCM10,CDC45,MKI67,ASPM,NCAPG,NUSAP1,ESPL1 and CDT1.Conclusion:These immune characteristics and hub genes may be correlated with the pathogenesis of RSV bronchiolitis in children,and the specific pathogenic mechanism deserves further investigation.

关 键 词：RNA测序 呼吸道合胞病毒 毛细支气管炎 关键基因 

分 类 号：R725.6[医药卫生—儿科]