摘要
本研究通过对呼吸道合胞病毒(RSV)毛细支气管炎患儿血白细胞进行RNA测序,探讨与发病机制及疾病严重度相关的生物学特征。本研究是一项病例对照研究,以2019年10月至2022年4月于温州医科大学附属第二医院儿童呼吸科住院临床诊断为毛细支气管炎,RSV抗原阳性和(或)RSV核酸阳性的87例患儿作为病例组,将病例组按病情分为轻、中、重三组,按有无特应征分为特应征组和无特应征组,选择同期40名健康体检儿童作为对照组,提取病例组和对照组儿童的全血白细胞RNA进行测序,分析数据得到差异表达基因(DEGs),再通过基因本体论(GO)注释、京都基因和基因组百科全书(KEGG)注释、蛋白质相互作用网络(PPI)构建筛选与发病机制、病情以及特应征相关的免疫生物学通路及基因。通过加权基因共表达网络分析(WGCNA)构建共表达网络,寻找与疾病严重度相关的潜在生物学指标。结果显示,病例组与对照组相比DEGs共有1782个,其中上调基因1586个,下调基因196个,这些DEGs的GO通路富集显示在分子功能中主要富集在过氧化物酶活性、氧化还原酶活性等过程,在细胞学组分中主要富集在细胞质囊泡腔、分泌颗粒腔中,在生物学过程中主要富集在中性粒细胞激活参与免疫反应、中性粒细胞脱颗粒、中性粒细胞激活等过程。DEGs的KEGG富集分析显示主要富集于病毒蛋白与细胞因子及其受体的相互作用信号通路。构建PPI网络后得到CCL2、IL-10、MMP9和JUN共4个处于核心位置的基因。不同病情组与对照组比较所得DEGs主要富集在逆行内源性大麻素信号转导和细胞凋亡通路上。WGCNA分析显示与血氧饱和度相关的棕色模块与病情关系最为密切,其基因主要富集在RNA解旋酶维甲酸诱导基因-I(RIG-I)样受体信号通路上。特应征组的特异性DEGs有230个,无特应征组的特异性DEGs有444个,KEGG富集分析结果显示两组均富集到NF-κB信号通路上,特应征不会导致RSV毛细支气管炎儿童免疫反应和转录组特征发生明显改变。综上,中性粒细胞激活、脱颗粒途径以及病毒蛋白与细胞因子及细胞因子受体的相互作用信号通路参与RSV毛细支气管炎宿主的免疫反应。CCL2、IL-10、MMP9和JUN基因可能与发病相关。在RIG-I样受体、细胞凋亡及内源性大麻素相关的信号通路中可能存在与疾病严重度相关的潜在生物学标志物。
To explore the biological characteristics related to the pathogenesis and severity of respiratory syncytial virus(RSV)bronchiolitis by RNA sequencing of white blood cells in children with RSV bronchiolitis.This study is a case-control study.A total of 87 children diagnosed with bronchiolitis and RSV antigen positive and/or RSV nucleic acid positive in the pediatric respiratory department of the Second Affiliated Hospital of Wenzhou Medical University from October 2019 to April 2022 were selected as the case group.The case group was divided into three groups based on the condition:mild,moderate,and severe,and there were two groups according to the presence or absence of atopic symptoms:the atopic group and the non-atopic group,forty healthy children in the same period were selected as the control group.The whole blood leukocyte RNA of the children in the case group and the control group was extracted for RNA sequencing,and the data were analyzed to obtain differentially expressed genes(DEGs).Then,the immunobiological pathways and genes related to the pathogenesis,disease condition,and atopy were screened through Gene Ontology(GO)annotation,Kyoto Gene and Genome Encyclopedia(KEGG)annotation,and protein interaction network(PPI)construction methods.Construct the weighted gene co-expression network analysis(WGCNA)module to identify potential biological indicators related to disease severity.Compared with the control group,the case group had a total of 1782 DEGs,including 1586 upregulated genes and 196 downregulated genes.The GO pathway enrichment of DEGs is mainly enriched in molecular functions such as peroxidase activity and oxidoreductase activity.In the cytological components,it is mainly enriched in cytoplasmic vesicle lumen and secretory granule lumen.In biological processes,it is mainly enriched in processes such as neutrophil activation involved in immune responses,neutrophil degranulation,and neutrophil activation.KEGG analysis is mainly concentrated in the signal pathway of the viral protein interaction with cytokine and cytokine receptor.A PPI network was constructed to screen four genes at the core position,including CCL2,IL-10,MMP9 and JUN.The DEGs obtained by comparing different disease groups with the control group are mainly enriched in retrograde endocannabinoid signaling and cell apoptosis pathways.WGCNA analysis showed that the brown module related to oxygen saturation was most closely related to the disease,and its gene was mainly enriched in the RNA helicase retinoic acid inducible gene-I(RIG-I)like receptor signal pathway.There are 230 specific DEGs in the atopic group and 444 in the non-atopic group.KEGG enrichment analysis results show that both groups are enriched to NF-κB signaling pathway,the characteristic does not cause significant changes in immune response and transcriptome characteristics in children with RSV bronchiolitis.In conclusion,neutrophil activation,degranulation pathway and signal pathway of interaction between viral protein and cytokine and cytokine receptor are involved in the immune response of RSV bronchiolitis host.CCL2,IL-10,MMP9 and JUN genes may be associated with the pathogenesis.They might be potential biomarkers related to disease severity in RIG-I like receptors,cell apoptosis,and endogenous cannabinoid related signaling pathways.
作者
王乐颖
乐伊莎
李海燕
刘振伟
翁婷婷
陈小芳
留佩宁
董琳
Wang Leying;Le Yisha;Li Haiyan;Liu Zhenwei;Weng Tingting;Chen Xiaofang;Liu Peining;Dong Lin(Department of Pediatric Pulmonology,the Second Affiliated Hospital and Yuying Children′s Hospital of Wenzhou Medical University,Wenzhou 325027,China;Institute of Genomic Medicine,Wenzhou Medical University,Wenzhou 325025,China;Department of Child Health Care Department,the Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University,Wenzhou 325027,China)
出处
《中华预防医学杂志》
CAS
CSCD
北大核心
2024年第1期71-80,共10页
Chinese Journal of Preventive Medicine
基金
浙江省基础公益研究计划(GF20H010020)
温州市基础性科研项目(Y20190437)
温州医科大学附属第二医院贺林新医学临床转化工作站科研基金重点项目(18331103)。
