Chen Huan, Liu Jinglan, Wu Yuqing, Jiang Li, Tang Mi, Wang Xin, Fang Xiaoling, Wang Xi
Department of Obstetrics and Gynecology, The Second XIANGYA Hospital Of Central South University, Changsha, China.
Department of Obstetrics and Gynecology, Zhu Zhou Hospital Affiliated to Xiangya school of medicine, CSU, Zhuzhou, China.
Front Surg. 2022 Aug 24;9:963850. doi: 10.3389/fsurg.2022.963850. eCollection 2022.
Congenital heart disease (CHD) is the most common type of birth defect. Most patients with CHD require surgery, and cardiopulmonary bypass (CPB) is the most common surgery performed.
The present study utilized weighted gene co-expression network analysis (WGCNA) to identify key inflammation genes after CPB for CHD. The GSE132176 dataset was downloaded from the Gene Expression Omnibus(GEO) database for WGCNA to identify the modules closely related to clinical traits. Disease enrichment, functional annotation and pathway enrichment were performed on genes in the module closely related to clinical traits using Enrichr and Metascape. Immune infiltration analysis was also performed on the training dataset using CIBERSORT. Finally, we identified hub genes using high gene significance (GS), high module members (MMs) and Cytoscape, and we verified the hub genes using an independent dataset and Western blot analysis.
WGCNA showed that the brown module with 461 genes had the highest correlation to CHD after CPB. Functional annotation and pathway enrichment analysis were performed using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses, which showed that genes in the brown module were enriched in inflammation-related pathways. In the disease enrichment analysis, genes in the brown module were enriched for inflammatory diseases. After the 30 most highly associated brown intramodular genes were screened, a protein-protein interaction network was constructed using the STRING online analysis website. The protein-protein interaction results were then calculated using 12 algorithms in the cytoHubba plugin of Cytoscape software. The final result showed that was the fundamental gene of post-CPB for CHD. Using another independent validation dataset (GSE12486), we confirmed that was significantly differentially expressed between preoperative and postoperative CPB (Wilcoxon, = 0.0079; T-test, = 0.006). In addition, expression was elevated in eosinophils, neutrophils, memory CD4 T cells and activated mast cells. Western blot analysis showed that the expression of CDKN1A protein was significantly higher postoperative CPB than preoperative CPB. Moreover, was mainly related to inflammation.
In summary, we found a relationship between and inflammation after CPB for congenital heart disease by WGCNA, experiments and various bioinformatics methods. Thus, maybe serve as a biomarker or therapeutic target for accurate diagnosis and treatment of inflammation after CPB in the future.
先天性心脏病（CHD）是最常见的出生缺陷类型。大多数CHD患者需要手术治疗，体外循环（CPB）是最常进行的手术。
本研究利用加权基因共表达网络分析（WGCNA）来识别CHD患者CPB术后的关键炎症基因。从基因表达综合数据库（GEO）下载GSE132176数据集用于WGCNA，以识别与临床特征密切相关的模块。使用Enrichr和Metascape对与临床特征密切相关模块中的基因进行疾病富集、功能注释和通路富集分析。还使用CIBERSORT对训练数据集进行免疫浸润分析。最后，我们使用高基因显著性（GS）、高模块成员度（MMs）和Cytoscape识别枢纽基因，并使用独立数据集和蛋白质印迹分析验证枢纽基因。
WGCNA显示，包含461个基因的棕色模块与CPB术后的CHD相关性最高。使用基因本体论（GO）和京都基因与基因组百科全书（KEGG）分析进行功能注释和通路富集分析，结果表明棕色模块中的基因在炎症相关通路中富集。在疾病富集分析中，棕色模块中的基因在炎症性疾病中富集。筛选出30个棕色模块内相关性最高的基因后，使用STRING在线分析网站构建蛋白质-蛋白质相互作用网络。然后使用Cytoscape软件的cytoHubba插件中的12种算法计算蛋白质-蛋白质相互作用结果。最终结果表明，[具体基因名称]是CHD患者CPB术后的关键基因。使用另一个独立验证数据集（GSE12486），我们证实[具体基因名称]在CPB术前和术后存在显著差异表达（Wilcoxon检验，P = 0.0079；T检验，P = 0.006）。此外，[具体基因名称]在嗜酸性粒细胞、中性粒细胞、记忆性CD4 T细胞和活化肥大细胞中表达升高。蛋白质印迹分析表明，CPB术后CDKN1A蛋白的表达明显高于术前。而且，[具体基因名称]主要与炎症相关。
综上所述，我们通过WGCNA、实验和各种生物信息学方法发现了[具体基因名称]与先天性心脏病CPB术后炎症之间的关系。因此，[具体基因名称]未来可能作为生物标志物或治疗靶点，用于CPB术后炎症的准确诊断和治疗。
