Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2021 Apr 26;9(5):471.
doi: 10.3390/biomedicines9050471.

MicroRNA Expression in Extracellular Vesicles from Nasal Lavage Fluid in Chronic Rhinosinusitis

Seungbin Cha  1 , Eun-Hye Seo  2 , Seung Hyun Lee  1   2 , Kyung Soo Kim  3 , Chung-Sik Oh  2   4 , Jong-Seok Moon  5 , Jin Kook Kim  2   6
Affiliations

MicroRNA Expression in Extracellular Vesicles from Nasal Lavage Fluid in Chronic Rhinosinusitis

Seungbin Cha et al. Biomedicines. .

Abstract

Extracellular vesicles (EVs) are nanovesicles of endocytic origin released by cells and found in human bodily fluids. EVs contain both mRNA and microRNA (miRNA), which can be shuttled between cells, indicating their role in cell communication. This study investigated whether nasal secretions contain EVs and whether these EVs contain RNA. EVs were isolated from nasal lavage fluid (NLF) using sequential centrifugation. EVs were characterized and EV sizes were identified by transmission electron microscopy (TEM). In addition, EV miRNA expression was different in the chronic rhinosinusitis without nasal polyp (CRSsNP) and chronic rhinosinusitis with nasal polyp (CRSwNP) groups. The Kyoto encyclopedia gene and genome database (KEGG) database was used to identify pathways associated with changed miRNAs in each analysis group. Twelve miRNAs were differentially expressed in NLF-EVs of CRS patients versus HCs. In addition, eight miRNAs were differentially expressed in NLF-EVs of CRSwNP versus CRSsNP patients. The mucin-type O-glycan biosynthesis was a high-ranked predicted pathway in CRS patients versus healthy controls (HCs), and the Transforming growth factor beta (TGF-β) signaling pathway was a high-ranked predicted pathway in CRSwNP versus CRSsNP patients. We demonstrated the presence of and differences in NLF-EV miRNAs between CRS patients and HCs. These findings open up a broad and novel area of research on CRS pathophysiology as driven by miRNA cell communication.

Keywords: extracellular vesicles; microRNAs; microarray analysis; nasal lavage fluid; sinusitis.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Phenotypic characterization of NLF-EVs. EV shape and size were analyzed by TEM. NLF, nasal lavage fluid; EV, extracellular vesicle; TEM, transmission electron microscopy.
Figure 2
Figure 2
Microarray and analysis of the results in CRS and healthy NLF-EVs. (A) Volcano plot showing the relationship between FC and statistical significance. The point of the plot indicates a differentially expressed miRNA with statistical significance. (B) Heat map diagram showing cluster analysis of differently expressed miRNAs. Heat maps and hierarchical clusters of selected differentially expressed miRNAs (p < 0.05). The sample clustering tree appears at the top, and the miRNA clustering tree is on the left. The color scale reflects the log2 signal strength and goes from green (low intensity) to black (medium intensity) to red (high intensity). The dendrogram at the top and left reflects the hierarchical similarity of the sample and miRNA individually. CRS, chronic rhinosinusitis; NLF, nasal lavage fluid; EV, extracellular vesicle; FC, fold change; miRNA, microRNA.
Figure 3
Figure 3
Top 7 KEGG biological pathways revealed by analyzing target miRNAs between NLF-EVs of CRS patients and HCs using DIANA tools. (A) Overexpressed miRNAs in NLF-EVs of CRS patients compared to HCs after adjustment of the outlier values according to the Chauvenet criterion (Student’s t-test; * p < 0.05; ** p < 0.01, ns > 0.05). (B) KEGG enrichment analysis of all DEGs with |FC| > 2 (cutoff). (C) Pathway enriched by up- and downregulated genes and sorted by the value of −log10(p-value). (D) Diagram of the mucin-type O-glycan biosynthesis. Five dysregulated miRNAs (miR-223-3p, miR-23a-3p, miR-15a-5p, miR-1285-3p, and miR-450a-1-3p) were associated with the mucin-type O-glycan biosynthesis by possibly regulating their potential gene targets. KEGG, Kyoto Encyclopedia Gene and Genome; miRNA, microRNA; NLF, nasal lavage fluid; EV, extracellular vesicle; CRS, chronic rhinosinusitis; HC, healthy control; DEG, differentially expressed gene; FC, fold change.
Figure 4
Figure 4
Top 10 KEGG biological pathways revealed by analyzing the target miRNAs between CRSsNP and CRSwNP NLF-EVs using DIANA tools. (A) Overexpressed miRNAs in NLF-EVs of CRS patients compared to HCs after adjustment of outlier values according to the Chauvenet criterion (Student’s t-test; * p < 0.05; ** p < 0.01; *** p < 0.001). (B) KEGG enrichment analysis of all DEGs with |FC| > 2 (cutoff). (C) Pathway enriched by up- and downregulated genes and sorted by the value of −log10(p-value). (D) Diagram of the TGF-β signaling pathway. Six dysregulated miRNAs (miR-1290, miR-548q, miR-548l, miR-1254, miR-890, and miR-548aa/548t) were associated with the TGF-β signaling pathway by possibly regulating their potential gene targets. KEGG, Kyoto Encyclopedia Gene and Genome; miRNA, microRNA; CRSsNP, chronic rhinosinusitis without nasal polyps; CRSwNP, chronic rhinosinusitis with nasal polyps; NLF, nasal lavage fluid; EV, extracellular vesicle; CRS, chronic rhinosinusitis; HC, healthy control; DEG, differentially expressed gene; FC, fold change; TGF-β, transforming growth factor-beta.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Wu G., Yang G., Zhang R., Xu G., Zhang L., Wen W., Lu J., Liu J., Yu Y. Altered microRNA Expression Profiles of Extracellular Vesicles in Nasal Mucus From Patients With Allergic Rhinitis. Allergy Asthma Immunol. Res. 2015;7:449–457. doi: 10.4168/aair.2015.7.5.449. - DOI - PMC - PubMed
    1. Lässer C., O’Neil S.E., Ekerljung L., Ekström K., Sjöstrand M., Lötvall J. RNA-containing Exosomes in Human Nasal Secretions. Am. J. Rhinol. Allergy. 2011;25:89–93. doi: 10.2500/ajra.2011.25.3573. - DOI - PubMed
    1. Taylor D.D., Gercel-Taylor C. MicroRNA signatures of tumor-derived exosomes as diagnostic biomarkers of ovarian cancer. Gynecol. Oncol. 2008;110:13–21. doi: 10.1016/j.ygyno.2008.04.033. - DOI - PubMed
    1. Nocera A.L., Miyake M.M., Seifert P., Han X., Bleier B.S. Exosomes mediate interepithelial transfer of functional P-glycoprotein in chronic rhinosinusitis with nasal polyps. Laryngoscope. 2017;127:E295–E300. doi: 10.1002/lary.26614. - DOI - PubMed
    1. Xuan L., Luan G., Wang Y., Lan F., Zhang X., Hao Y., Zheng M., Wang X., Zhang L. MicroRNAs regulating mucin type O-glycan biosynthesis and transforming growth factor β signaling pathways in nasal mucosa of patients with chronic rhinosinusitis with nasal polyps in Northern China. Int. Forum Allergy Rhinol. 2019;9:106–113. doi: 10.1002/alr.22230. - DOI - PubMed

LinkOut - more resources