Serum exosomal miR-141-3p and miR-3679-5p levels associated with endotype and postoperative recurrence in chronic rhinosinusitis with nasal polyps

Abstract

Background: Chronic rhinosinusitis with nasal polyps (CRSwNP) is a chronic inflammatory disease. Exosomes were involved in different inflammatory diseases, but their roles in CRSwNP were poorly explored.

Method: We collected serum samples from 8 CRSwNP patients and 8 healthy controls (HC) and isolated their exosomes. MiRNA sequencing was performed for the exosome samples and differentially expressed miRNAs were identified. The top 3 differentially expressed exosomal miRNAs were confirmed in 2 validation cohorts, and their diagnostic values, predictive values for eosinophilic endotype, and recurrence were evaluated.

Results: Distinctive serum exosomal miRNA profiles were observed between CRSwNP and HC groups. Reverse transcription-polymerase chain reaction results in the first validation cohort revealed that serum exosomal miR-141-3p levels were increased, and miR-18a-5p and miR-3679-5p levels were decreased in the CRSwNP group compared to the HC group. These 3 miRNAs were further validated in the second validation cohort, and the results showed that miR-141-3p levels were elevated and miR-3679-5p levels were reduced in the serum exosomes in the eosinophilic CRSwNP group in comparison with the non-eosinophilic CRSwNP group. Receiver operating characteristic (ROC) curves highlighted that exosomal miR-141-3p and miR-3679-5p exhibited promising values for predicting the eosinophilic endotype. The patients in the second cohort were followed up for 2 years, and categorized into recurrence and non-recurrence groups. The serum exosomal miR-141-3p levels were increased and miR-3679-5p levels were reduced in the recurrence group in comparison with the non-recurrence group. ROC curves and Kaplan-Meier survival analysis revealed significant associations between the levels of exosomal miR-141-3p and miR-3679-5p and the risk of postoperative recurrence.

Conclusions: This study identified unique miRNA expression patterns in serum exosomes of CRSwNP patients. Circulating exosomal miR-141-3p and miR-3679-5p emerged as novel biomarkers for diagnosing CRSwNP, predicting the eosinophilic endotype, and forecasting postoperative recurrence.

Keywords: Biomarkers; Exosomes; Nasal polyps; Recurrence.

Fig. 1

Landscape of exosomal miRNA profiles between HC and CRSwNP groups in the discovery cohort. (A) The heat map shows the exosomal miRNA profiles between HC and CRSwNP groups. (B) The volcano plot of differentially expressed serum exosomal miRNAs. (C) The top 10 most significant KEGG pathway terms of target genes. HC, healthy control; CRSwNP, chronic rhinosinusitis with nasal polyps. KEGG, kyoto encyclopedia of genes and genomes

Fig. 2

Validation of the top 3 most differentially expressed serum exosomal miRNAs in the first validation cohort. (A–F) comparison of the expressions of serum exosomal miRNAs between the HC and CRSwNP groups. (G–L) ROC curves show the diagnostic values of serum exosomal miRNAs for CRSwNP. HC, healthy control; CRSwNP, chronic rhinosinusitis with nasal polyps. ROC, receiver operating characteristic. ns, no significance; ∗∗P < 0.01; ∗∗∗P < 0.001

Fig. 3

Exploration of expression levels of 3 candidate miRNAs in the serum EXO between the neCRSwNP and eCRSwNP groups in the second validation cohort. (A–C) Comparisons of serum exosomal miR-141-3p, miR-18a-5p, and miR-3679-5p expressions between the neCRSwNP and eCRSwNP groups. (D-F) ROC curves show the predictive values of serum exosomal miR-141-3p, miR-18a-5p, and miR-3679-5p for eCRSwNP. EXO, exosome; neCRSwNP, non-eosinophilic chronic rhinosinusitis with nasal polyps; eCRSwNP, eosinophilic chronic rhinosinusitis with nasal polyps. ns, no significance; ∗∗∗P < 0.001

Fig. 4

Exploration of expression levels of 3 candidate miRNAs in the serum EXO between the non-recurrence and recurrence groups. (A–C) Comparisons of serum exosomal miR-141-3p, miR-18a-5p, and miR-3679-5p expressions between the 2 groups. (D–F) The association between serum exosomal miR-141-3p, miR-18a-5p, and miR-3679-5p expressions and recurrence in the neCRSwNP and eCRSwNP groups. EXO, exosome; neCRSwNP, non-eosinophilic chronic rhinosinusitis with nasal polyps; eCRSwNP, eosinophilic chronic rhinosinusitis with nasal polyps. ns, no significance; ∗P < 0.05; ∗∗P < 0.001; ∗∗∗P < 0.001

Fig. 5

The predictive values of serum exosomal miR-141-3p, miR-18a-5p, and miR-3679-5p for postoperative recurrence. (A–C) ROC curves; (D–F) Kaplan-Meier survival analysis. ROC, receiver operating characteristic; AUC, the area under the curve

Fig. S1

Characterization and identification of serum EXO in HCs and CRSwNP patients in the discovery cohort. (A) Electron microscopic images of serum EXO. Scale bar = 100 nm. (B) Nanoparticle tracking analysis of the size distribution of serum EXO. (C) Immunoblotting analysis of CD63 and CD81 in EXO and serum without EXO. EXO, exosome; HC, healthy control; CRSwNP, chronic rhinosinusitis with nasal polyps.