A Study of Nasal Epithelial Cell Membrane in Patients with Chronic Rhinosinusitis without Nasal Polyps Using a Fluorescent Probe

Abstract Objectives: The hippo-yes-associated protein (YAP) pathway plays an important role in epithelial cell proliferation and inflammation in chronic rhinosinusitis with nasal polyps (CRSwNP). However, the underlying mechanisms remain unclear. This study intends to investigate the role of YAP and the nuclear factor kappa-B (NF-κB) signalling pathway in nasal epithelial cell proliferation and the expression of epithelium-derived cytokines in CRSwNP.Methods: The expression levels of YAP, TEAD1, Ki-67, and NF-κB in sinonasal mucosa, primary nasal epithelial cells (NPECs), and human nasal epithelial RPMI 2650 cells were detected by RT-qPCR and immunoblotting. NPECs were cultured and treated with verteporfin (VP), a selective YAP inhibitor, YAP shRNA or BAY 11-7082, a small molecule inhibitor of NF-κB. The relationship between cell proliferation and hippo pathway activity was explored using a cell counting kit-8 (CCK-8) assay, 5(6)-carboxyfluorescein diacetate succinimidyl ester (CFSE) labelling and colony formation assay. The cell cycle and apoptosis were examined through flow cytometry (FCM) assay. The epithelium-derived cytokines including interleukin (IL-) 33, IL-25 and thymic stromal lymphopoietin(TSLP) were detected by RT-qPCR.Results: The hippo pathway effector YAP, Ki-67, p65 NF-κB, and cyclin D1were significantly increased in CRSwNP compared with control mucosa; which was accompanied by overexpression of interleukin (IL)-33, IL-25, and thymic stromal lymphopoieth (TSLP). Pharmaceutical inhibition of YAP by VP suppressed cell proliferation of RPMI 2650 cells by blocking cell cycle progression at G0/G1 without inducing obvious cell apoptosis. Furthermore, lentiviral transfection-mediated knockdown of hippo pathway activity reduced the expression of IL-33,,IL-25, TSLP as well as p65 NF-κB in RPMI 2650 cells. Downregulation of NF-κB pathway with BAY 11-7082 in NPECs could decrease the mRNA level of TSLP, IL-33 and IL-25 accordingly.Conclusions: Inhibition of hippo pathway suppressednasal epithelial cell proliferation and declined the expression of epithelium-derived cytokines IL-33 and IL-25 and TSLP expression via the NF-κB signalling pathway in NPECs.

Download Full-text

Pathogenesis of chronic rhinosinusitis with nasal polyps: role of IL-6 in airway epithelial cell dysfunction

Journal of Translational Medicine ◽

10.1186/s12967-020-02309-9 ◽

2020 ◽

Vol 18 (1) ◽

Cited By ~ 1

Author(s):

Emilie Bequignon ◽

David Mangin ◽

Justine Bécaud ◽

Jennifer Pasquier ◽

Christelle Angely ◽

...

Keyword(s):

Epithelial Cell ◽

Chronic Rhinosinusitis ◽

Nasal Polyps ◽

Airway Epithelial Cell ◽

Airway Epithelial ◽

Cell Dysfunction

Download Full-text

Contribution of Epithelial Cell Dysfunction to the Pathogenesis of Chronic Rhinosinusitis with Nasal Polyps

American Journal of Rhinology and Allergy ◽

10.1177/1945892419868588 ◽

2019 ◽

Vol 33 (6) ◽

pp. 782-790 ◽

Cited By ~ 6

Author(s):

Michael Wynne ◽

Carl Atkinson ◽

Rodney J. Schlosser ◽

Jennifer K. Mulligan

Keyword(s):

Vitamin D ◽

T Cells ◽

Dendritic Cells ◽

Epithelial Cell ◽

Chronic Rhinosinusitis ◽

Airway Epithelium ◽

Nasal Polyps ◽

Mucociliary Clearance ◽

Innate And Adaptive Immunity ◽

Cell Dysfunction

Background In the past, the airway epithelium was thought to be primarily an inert physical barrier. We now know that the upper airway epithelium plays a critical role in both innate and adaptive immunity, and that epithelial dysfunction is strongly associated with inflammatory airway disease. The pathogenesis of chronic rhinosinusitis is poorly understood, but growing evidence supports a key role for the airway epithelium in the pathophysiology of the disease. Objective The purpose of this study is to explore our current understanding of how dysfunction in human sinonasal epithelial cells (HSNECs) contributes to the pathogenesis of chronic rhinosinusitis with nasal polyps (CRSwNP) and to examine how current and developing therapies affect epithelial cell functions. Methods A literature review of papers published in English pertaining to epithelial cell dysfunction in patients with CRSwNP was performed using the PubMed database. The search utilized combinations of the following key words: sinusitis, polyps, epithelium, pathophysiology, barrier function, dendritic cells, eosinophils, T cells, complement, mucociliary clearance, vitamin D, cytokines, chemokines, taste receptors, steroids, saline, and therapy. Results HSNEC mucociliary clearance, barrier function, secretion of cytokines, influence on dendritic cells, influence on T-cells, regulation of eosinophils, vitamin D metabolism, complement production, and taste receptor function are altered in patients with CRSwNP and contribute to the pathogenesis of the disease. Current therapies utilized to manage CRSwNP counteract the effects of HSNEC dysfunction and relieve key symptoms of the disease. Conclusion HSNECs are key players in both innate and adaptive immunity, and altered epithelial functions are closely intertwined with the pathogenesis of CRSwNP. Our review supports further investigation of altered HSNEC function in patients with CRSwNP and supports development of novel epithelial-targeted therapies for its management.

Download Full-text