Expression of Apoptosis-Related Biomarkers in Inflamed Nasal Sinus Epithelium of Patients with Chronic Rhinosinusitis with Nasal Polyps (CRSwNP)—Evaluation at mRNA and miRNA Levels

Abstract

In chronic upper respiratory tract diseases, increased cell proliferative activity is observed, which is coordinated by BCL-2 proteins and small non-coding RNAs. This study aimed to determine the expression of critical apoptosis markers at the mRNA and miRNA levels in patients with chronic rhinosinusitis with nasal polyps (CSRwNP). The study group consisted of ten patients with CSRwNP and ten healthy subjects. To detect in situ apoptosis in the maxillary sinus mucosa, TUNEL staining was performed. The expression of transcripts was determined by RT-qPCR and included the detection of markers associated with cell survival and apoptosis, i.e., BAX, p53, p21, CASP3, CASP9, c-MYC, CCND1, BRIC5, and APAF1. Levels of miR-17-5p, miR-145-5p, miR-146a-5p, and miR-203a-3p were also measured by RT-qPCR. The obtained results indicated increased apoptosis determined by a TUNEL assay in CSRwNP patients and accompanied by an increased expression of BAX, P21, P53, CASP3, CASP9, c-MYC, and APAF-1 transcripts and decreased mRNA levels of BCL-2 and BIRC5. Furthermore, the nasal sinus epithelium of patients with CSRwNP showed increased levels of miR-203a-3p while also showing a decreased expression of miR-17-5p and miR-145-5p. Our results showed that pro-apoptotic transcripts detected at mRNA and miRNA levels might be involved in the pathogenesis of chronic sinusitis with polyps. The identification of those key molecular mediators may be applicable for the specific diagnostic and/or development of targeted therapies for chronic sinusitis with polyps.

MicroRNAs: Potential Biomarkers of Disease Severity in Chronic Rhinosinusitis with Nasal Polyps

“…It is possible that with better defined patient categories, miR-203a-3p could be a tool for predicting the atopic status of CRSwNP patients. On the other hand, a recently published study from Poland [ 35 ] described higher values of miR-203a-3p in CRSwNP patients, compared to controls, yet the RT-qPCR analysis was performed on the maxillary sinus mucosa of 10 CRSwNP patients and not on nasal polyps. Additional research would be warranted for clarifying the association between miR-203a-3p and nasal polyposis.…”

Epigenetic modifications in chronic rhinosinusitis with and without nasal polyps

“…In addition, miR-146a ( Xia et al, 2015 ), miR-614 ( Shin et al, 2020 ), and some other DE-miRs are also upregulated in CRS samples, highly through the pathway of mucin-type O-glycan biosynthesis ( Cha et al, 2021 ). Meanwhile, miR-3178, miR-585-3p, miR-3146, miR-320e, miR-142-3p, miR-154, miR-223, miR-205-5p, miR-222-3p, miR-378a-3p, miR-449b-5p, miR-22-3p, miR-21, miR-203a-3p, miR-677, miR-1037, and miR-79 are also overexpressed in CRSwNP samples ( Li et al, 2019c ; Xuan et al, 2019 ; Zhang et al, 2020b ; Bu et al, 2021 ; Qing et al, 2021 ; Silveira et al, 2021 ; He et al, 2022 ; Morawska-Kochman et al, 2022 ). Among them, miR-142-3p may regulate the inflammatory response through the lipopolysaccharide (LPS)-Toll-like receptor (TLR)-tumor necrosis factor-α (TNF-α) signaling pathway ( Qing et al, 2021 ); miR-3146 and miR-320e act in the mucin type O-glycan biosynthesis pathway in CRSwNP by targeting N-acetylgalactosaminyltransferase-like 6 (GALNTL6), N-acetylgalactosaminyltransferase 8 (GALNT8), and alpha-2,3-sialyltransferase 1 (ST3GAL1) genes ( Xuan et al, 2019 ); miR-223 plays in the mitogen-activated protein kinase (MAPK) signaling pathway ( Bu et al, 2021 ); miR-22-3p inhibits vascular endothelial (VE)-cadherin expression to regulate vascular permeability after binding to the specific site in the 3′-UTR of the human VE-cadherin mRNA ( Zhang et al, 2020b ); the TGF-β1-miR-21-PTEN-Akt axis may propel the EMT of CRSwNP ( Li et al, 2019c ); miR-677, miR-1037, and miR-79 may post-transcriptionally regulate gene expression in the Hippo signaling pathway ( He et al, 2022 ).…”

“…Additional analysis reveals that by repressing epidermal growth factor receptor (EGFR), miR-146a modulates mucin-5AC (MUC5AC) induced by human neutrophil elastase hypersecretion, and EGFR is a target gene of miR-146a ( Yan et al, 2020 ). What’s more, miR-146a-5p expression shows no significant difference between healthy subjects and CRSwNP patients ( Morawska-Kochman et al, 2022 ). The reasons for these conflicting results are unclear and need further investigation.…”

“…In addition, the receiver operating characteristic (ROC) curve analysis has shown the area under the curve (AUC) of miR-145-5p in NPs is .8690, with a p -value statistically significant, strongly supporting its involvement in the genesis of CRSwNP ( Yu et al, 2021b ). Besides, miRNAs are also downregulated although the mechanism yet-to-be explored, such as miR-92a, miR-26b, miR-708-5p, and miR-126-3p in CRS ( Ma et al, 2015 ; Xia et al, 2015 ), miR-132-3p, miR-27b-3p, miR-17-5p, and miR-125a in CRSwNP ( Yu et al, 2021b ; Chen et al, 2022 ; Morawska-Kochman et al, 2022 ), miR-26a in ECRSwNP ( Zhang et al, 2014 ), and miR-1290 in CRSsNP ( Ma et al, 2015 ) ( Table 2 ).…”

Circulating levels of Bcl-2 and its expression in the nasal mucosa of patients with chronic rhinosinusitis

“…Features of Bcl-2 expression in cells of sinonasal mucosa and the role of this anti-apoptotic protein in CRS have been under investigation with some promising results reported recently [17]. We have hypothesized that Bcl-2 expression may be altered in CRS.…”