Wang L Y, Le Y S, Li H Y, Liu Z W, Weng T T, Chen X F, Liu P N, Dong L
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.
Zhonghua Yu Fang Yi Xue Za Zhi. 2024 Jan 6;58(1):71-80. doi: 10.3760/cma.j.cn112150-20230126-00057.
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 nonatopic 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 1 782 DEGs, including 1 586 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 and . 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 nonatopic 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. and 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.
通过对呼吸道合胞病毒(RSV)细支气管炎患儿白细胞进行RNA测序,探索与RSV细支气管炎发病机制及严重程度相关的生物学特征。本研究为病例对照研究。选取2019年10月至2022年4月在温州医科大学附属第二医院儿科呼吸科诊断为细支气管炎且RSV抗原阳性和/或RSV核酸阳性的87例患儿作为病例组。病例组根据病情分为轻度、中度和重度三组,根据是否有特应性症状分为特应性组和非特应性组,同期选取40例健康儿童作为对照组。提取病例组和对照组患儿的全血白细胞RNA进行RNA测序,并对数据进行分析以获得差异表达基因(DEG)。然后,通过基因本体论(GO)注释、京都基因与基因组百科全书(KEGG)注释和蛋白质相互作用网络(PPI)构建方法,筛选出与发病机制、疾病状况和特应性相关的免疫生物学途径和基因。构建加权基因共表达网络分析(WGCNA)模块,以识别与疾病严重程度相关的潜在生物学指标。与对照组相比,病例组共有1782个DEG,其中上调基因1586个,下调基因196个。DEG的GO通路富集主要集中在过氧化物酶活性和氧化还原酶活性等分子功能上。在细胞成分中,主要富集在细胞质囊泡腔和分泌颗粒腔中。在生物学过程中,主要富集在免疫反应中涉及的中性粒细胞活化、中性粒细胞脱颗粒和中性粒细胞活化等过程中。KEGG分析主要集中在病毒蛋白与细胞因子和细胞因子受体相互作用的信号通路中。构建PPI网络以筛选位于核心位置的四个基因,包括 和 。通过将不同疾病组与对照组比较获得的DEG主要富集在内源性大麻素逆行信号通路和细胞凋亡途径中。WGCNA分析表明,与血氧饱和度相关的棕色模块与疾病关系最为密切,其基因主要富集在RNA解旋酶视黄酸诱导基因-I(RIG-I)样受体信号通路中。特应性组有230个特异性DEG,非特应性组有444个。KEGG富集分析结果表明,两组均富集到NF-κB信号通路,这一特征在RSV细支气管炎患儿中不会引起免疫反应和转录组特征的显著变化。总之,中性粒细胞活化、脱颗粒途径以及病毒蛋白与细胞因子和细胞因子受体相互作用的信号通路参与了RSV细支气管炎宿主的免疫反应。 和 基因可能与发病机制有关。它们可能是RIG-I样受体、细胞凋亡和内源性大麻素相关信号通路中与疾病严重程度相关的潜在生物标志物。