Assessment Of The Upper And Lower Airway Unity In Patients With Chronic Rhinosinusitis With Nasal Polyps Via Proteomics Combined With Metabolomics

BACKGROUND AND OBJECTIVE Based on the united airway hyposesis,chronic rhinosinusitis with nasal polyps(CRSw NP)and asthma may share the same pathogenesis.However,there is a lack of convincing research evidence.Because of this,we tried to establish expression profiles of CRSw NP and asthma via proteomics and metabolomics.According to expression profile analyses,it could be able to identify the extent of similarity and to determine the unity of upper and lower airway pathogenesis.METHODS Six patients with CRSw NP,6 patients suffering from CRSw NP with asthma,and another 6 healthy people were recruited in this study.Nasal lavage fluid(NLF)and induced sputum were obtained from each subject,and cells were extracted from NLF and induced sputum,respectively.Independent data acquisition(DIA)and untargeted metabolomics were operated to analyze the expression profiles of proteins and metabolites,followed by Kendall ’s tau,a method for analyzing the correlations of proteins and metabolites expression profiles between upper and lower airways,respectively.According to abovementioned analyses,trend clustering modules and WGCNA modules were established to determine the similarity between upper and lower airways.Afterwards,Z-scores based on Module Preservation function calculations were obtained to assess the conservative degree of upper and lower airways.KEGG was performed to seek the differential expressions of proteins and metabolites between groups for further analysis.RESULTS 2444 and 2555 proteins were found in NLF and induced sputum,respectively,via DIA analysis.2182 proteins co-expressed in upper and lower airways,and the correlation coefficient is 0.68.KEGG analysis revealed that the metabolic pathways associated with CRSw NP or asthma include the followings,tight junction,ribosome,phagosome,neutrophil extracellular trap and endocytosis.Further analysis discovered a module mostly related to upper and lower airways,involving 730 proteins.At the metabolic level,LC-MS/MS confirmed 1535 metabolites,(including 1063 cationic model metabolites and 472 anionic model metabolites),with a correlation coefficient of 0.53 between upper and lower airways.Similar to proteinomics,WGCNA analysis discovered a mostly relevant module involving 218 metabolites.Joint analysis revealed that both the biosynthesis of unsaturated fatty acids and retrograde endogenous cannabinoid signal transduction.At the protein level,immune pathways such as coenzyme and aromatic hydrocarbon biosynthesis and asthma pathway play a role both in upper and lower airways in CRSw NP;asthma pathway and iron death pathway play a role both in upper and lower airways in CRSw NP with asthma;endocytosis,autophagy and iron death are associated with the development of asthma and the progress of asthma promoting CRSw NP.The expression of Ig E in the upper airway of CRSw NP&Asthma VS Control group is higher than that in CRSw NP VS Control,suggesting that Ig E in NLF might be a marker for assessing the occurrence of asthma in patients with CRSw NP.At the metabolic level,tyrosine and Ltryptophan involving metabolic pathways have been found to be associated with airway inflammation.CONCLUSIONS It was found that the components of proteins and metabolites were obviously conservative in upper and lower airways,confirming theory of united airway.This study discovered that proteins in the asthma pathway and iron death pathway are inducing factors leading of CRSw NP and Asthma.Ig E in NLF could indicate the occurrence of asthma.