The Expression Of EGF And AZGP1 Are Associated With Clinical Characteristics In Patients With Chronic Rhinosinusitis With Nas

Background and objectiveChronic rhinosinusitis(CRS)is a common inflammatory disease of the upper respiratory tract,which can be divided into Chronic rhinosinusitis with nasal polyps(CRSwNP)and Chronic rhinosinusitis without nasal polyps(CRSsNP).CRSwNP has a higher incidence,but its pathogenesis is still not completely clear and its treatment is limited due to the complexity of its occurrence and development.This study aimed to identify new biomarkers involved in the pathogenesis of CRSwNP by combined analysis of multiple transcriptome sequencing and correlated them with clinical symptoms to provide new strategies for the diagnosis and treatment of CRSwNP.MethodsThe GSE19428,GSE36830,and GSE23552 datasets were obtained from the Gene Expression Omnibus(GEO)database,the differentially expressed genes(DEGs)of CRSwNP were determined using R software,and the intersecting differentially expressed genes were obtained by the Draw Venn Diagram for gene ontology(GO)and Kyoto Encyclopedia of Genomes and Genomes(KEGG)enrichment analysis and were imported into STRING database to construct protein-protein interaction(PPI)network and further investigate the function of DEGs.Then,to detect the expression of the hub gene,nasal specimens from 90 CRSwNP patients and 45 normal controls were collected to perform qRT-PCR and receiver operator characteristic curve(ROC)analysis were applied.Next,according to the degree of eosinophilic infiltration,90 CRSwNP patients were classified into eosinophilic chronic rhinosinusitis with nasal polyps(eCRSwNP)and non-Eosinophilic chronic rhinosinusitis with nasal polyps(Non-eCRSwNP),and the differences in hub gene expression between the two groups were analyzed.Furthermore,immunohistochemical staining(IHC)of hub genes was performed to detect the protein expression levels of hub genes.Finally,correlation analysis of hub gene expression levels with clinical subjective and objective scores such as VAS,SNOT-22 score and Lund-Mackay CT score was performed.ResultsAfter screening and incorporating three GEO datasets(GSE19428,GSE36830,GSE23552),68 intersecting differentially expressed genes were obtained,including 8 upregulated genes and 60 down-regulated genes.GO functional analysis of the intersecting differentially expressed genes was performed,and they were mainly enriched in the positive regulation of phosphatidylinositol 3-kinase signaling,positive regulation of ERK1 and ERK2 cascade,transforming growth factor β receptor signaling pathway,negative regulation of cAMP-dependent protein kinase activity,Salivary secretion,Cytokine-cytokine receptor interaction and other pathways,which were involved in epithelial-mesenchymal transition(EMT).2 sub-networks with 6 hub genes were identified from the PPI network.Except PDGFC,the mRNA expression levels of EGF,ERBB4,AZGP1,CRISP3,and PIP were validated significantly down-regulated in CRSwNP.The ROC curve showed these 5 hub genes performed well with the area under the curves(AUC)are 0.7386,0.7342,0.8872,0.817,and 0.8367,respectively,and the AUC for the combination of the five genes was 0.8941.In the five hub genes,only the mRNA expression levels of EGF and AZGP1 were lower in eCRSwNP than in Non-eCRSwNP.IHC staining showed that the protein levels of EGF and AZGP1 in NP were significantly lower than in normal controls and showed a significant positive correlation with mRNA levels.In addition,according to the mRNA level of EGF,the patients with CRSwNP were divided into EGF low expression group and EGF high expression group.The analysis revealed that the VAS scores of nasal congestion,rhinorrhea,sneezing,nasal itching and eye itching symptoms were more severe in the EGF low expression group,and the rhinologic scores,psychological scores and total SNOT-22 scores were significantly higher than in the high expression group.CRSwNP patients were divided into AZGP1 low expression group and AZGP1 high expression group according to the mRNA expression level of AZGP 1,similarly,and it was found that VAS scores of nasal congestion,rhinorrhea,sneezing and nasal itching were significantly higher in the AZGP 1 low expression group than in the high expression group,and rhinologic scores in the SNOT-22 questionnaires were significantly higher than in the high expression group.Conclusion:In this study,we identified five hub genes EGF,ERBB4,AZGP1,CRISP3 and PIP,which have moderate diagnostic efficacy,Further analysis revealed that EGF and AZGP1 were differentially expressed between eCRSwNP and Non-eCRSwNP,and the VAS scores of nasal congestion,rhinorrhea,sneezing,nasal itching and eye itching,nasal symptoms SNOT-22,psychological symptoms SNOT-22 and total SNOT-22 scores were higher in the EGF low expression group,and the VAS scores of nasal congestion,rhinorrhea,sneezing,nasal itching,rhinologic scores in the SNOT-22 questionnaires were higher in the AZGP1 low expression group.Thus EGF and AZGP1 may play a potential role in the progression of type 2 inflammation and the development of CRSwNP.