Identification of key signaling pathways and hub genes related to immune infiltration in Kawasaki disease with resistance to intravenous immunoglobulin based on weighted gene co-expression network analysis

Abstract

Background: Kawasaki disease (KD) is an acute vasculitis, that is, the leading cause of acquired heart disease in children, with approximately 10%–20% of patients with KD suffering intravenous immunoglobulin (IVIG) resistance. Although the underlying mechanism of this phenomenon remains unclear, recent studies have revealed that immune cell infiltration may associate with its occurrence.Methods: In this study, we downloaded the expression profiles from the GSE48498 and GSE16797 datasets in the Gene Expression Omnibus database, analyzed differentially expressed genes (DEGs), and intersected the DEGs with the immune-related genes downloaded from the ImmPort database to obtain differentially expressed immune-related genes (DEIGs). Then CIBERSORT algorithm was used to calculate the immune cell compositions, followed by the WGCNA analysis to identify the module genes associated with immune cell infiltration. Next, we took the intersection of the selected module genes and DEIGs, then performed GO and KEGG enrichment analysis. Moreover, ROC curve validation, Spearman analysis with immune cells, TF, and miRNA regulation network, and potential drug prediction were implemented for the finally obtained hub genes.Results: The CIBERSORT algorithm showed that neutrophil expression was significantly higher in IVIG-resistant patients compared to IVIG-responsive patients. Next, we got differentially expressed neutrophil-related genes by intersecting DEIGs with neutrophil-related module genes obtained by WGCNA, for further analysis. Enrichment analysis revealed that these genes were associated with immune pathways, such as cytokine-cytokine receptor interaction and neutrophil extracellular trap formation. Then we combined the PPI network in the STRING database with the MCODE plugin in Cytoscape and identified 6 hub genes (TLR8, AQP9, CXCR1, FPR2, HCK, and IL1R2), which had good diagnostic performance in IVIG resistance according to ROC analysis. Furthermore, Spearman’s correlation analysis confirmed that these genes were closely related to neutrophils. Finally, TFs, miRNAs, and potential drugs targeting the hub genes were predicted, and TF-, miRNA-, and drug-gene networks were constructed.Conclusion: This study found that the 6 hub genes (TLR8, AQP9, CXCR1, FPR2, HCK, and IL1R2) were significantly associated with neutrophil cell infiltration, which played an important role in IVIG resistance. In a word, this work rendered potential diagnostic biomarkers and prospective therapeutic targets for IVIG-resistant patients.