Transcriptome Difference Analysis And Functional Study Of Chronic Nasal-sinusitis With Nasal Polyps Of Acidophilic And Non-eosinophilic

Background:Chronic rhinosinusitis with nasal polyps?CRSwNP?is one of the most prevalent chronic diseases with a heavy socioeconomic burden,characterized by persistent inflammation of sinonasal mucosa.CRSwNP is a heterogeneous disease,which can be divided into eosinophilic CRSwNP?ECRSwNP?and noneosinophilic CRSwNP?non-ECRSwNP?according to the number of eosinophils in the histopathology examination.The two phenotypes have different responses to corticosteroid and different prognosis,such as recurrence rate after surgery.But the pathogenesis of the differences between the two phenotypes remains unclear,let alone choose personalized therapeutic regimen for every patient.The aim of this study was to delineate a more complete transcriptome picture and identify key pathways that are dysregulated in eosinophilic and noneosinophilic CRSwNP.We used next-generation high-throughput RNA sequencing?RNA-Seq?to analyzed the profiles of gene expression in ECRSwNP,non-ECRSwNP and normal control subjects.A global vision of the complex transcriptome network of eosinophilic and non-eosinophilic CRSwNP will provide new insights into the underlying pathogenesis of CRSwNP.We believe these findings will be helpful in developing personalized therapeutic strategies.PART I The Investigation of the Difference of mRNA and IncRNA Expression Profile between Eosinophilic and Noneosinophilic CRSwNPObjective:To explore the mechanisms underlying the pathogenesis of ECRSwNP and non-ECRSwNP by global transcriptome analysis.Methods:RNA-Seq was applied to analyze the transcriptome profiles including mRNAs and IncRNAs in patients with ECRSwNP,non-ECRSwNP,and control subjects.The reliability of RNA-Seq results were verified by qRT-PCR in an independent large cohort of patients.The functions of differentially expressed genes were analyzed using bioinformatics.Results:A total of 1917 novel IncRNAs and 280 known IncRNAs were identified in this study.The IncRNA transcripts we detected displayed distinct genomic features compared with mRNA transcripts.The reliability of RNA-Seq results were validated by qRT-PCR showing that expression levels of five random dysregulated IncRNAs were consistent with the RNA-Seq data.The differentially expressed genes,such as CCL13 and CXCL9,may play an important role in the pathogenesis of ECRSwNP and non-ECRSwNP.Conclusion:LncRNAs existed in CRSwNP and were independent transcripts encoded from genomic regions with active transcriptional activity.RNA-Seq revealed CRSwNP and non-ECRSwNP possess distinct transcriptome profiles and provided us with a candidate reservoir of potential mRNAs and IncRNAs which may play a vital role in the pathogenesis of ECRSwNP and non-ECRSwNP.PART ? The Investigation of the Function and Mechanism of Differentially Expressed mRNAs and IncRNAs between Eosinophilic and Noneosinophilic CRSwNPObjective:To explore the mechanisms underlying the pathogenesis of ECRSwNP and non-ECRSwNP by analyzing the differentially and commonly expressed genes.Methods:The functions of dysregulated genes were analyzed by Gene ontology and KEGG pathway enrichment analysis.The functions of differentially expressed IncRNA were predicted by their cis or trans regulated target protein-coding genes.In situ RNA hybridization and qRT-PCR were performed to analyze the relationship between IncRNA XLOC₀10280 and its cis regulated target CCL18.Results:The differentially expressed genes between ECRSwNP and non-ECRSwNP were mainly enriched in inflammatory and immune response and extracellular microenvironment change.The predicted functions of dysregulated IncRNAs were highly consistent with those of dysregulated mRNAs,indicating IncRNAs play an important role in regulating the expression of mRNAs.LncRNA XLOC₀10280 and its cis regulated target CCL18 both specifically highly expressed in ECRSwNP and had a high Pearson correlation coefficient,suggesting XLOC₀10280 may play a role in eosinophilic inflammation by regulating CCL18 levels.Conclusion:The inflammatory and immune response and extracellular microenvironment change are two key characteristics of CRSwNP pathophysiology.LncRNAs can regulate the expression level of mRNAs of CRSwNP,and XLOC₀10280 may play a crucial role in eosinophilic inflammation of ECRSwNP by regulating its cis regulated target CCL18.