Host Antiviral Response Suppresses Ciliogenesis and Motile Ciliary Functions in the Nasal Epithelium

Abstract

BackgroundRespiratory viral infections are one of the main drivers of development and exacerbation for chronic airway inflammatory diseases. Increased viral susceptibility and impaired mucociliary clearance are often associated with chronic airway inflammatory diseases and served as risk factors of exacerbations. However, the links between viral susceptibility, viral clearance, and impaired mucociliary functions are unclear. Therefore, the objective of this study is to provide the insights into the effects of improper clearance of respiratory viruses from the epithelium following infection, and their resulting persistent activation of antiviral response, on mucociliary functions.MethodsIn order to investigate the effects of persistent antiviral responses triggered by viral components from improper clearance on cilia formation and function, we established an in vitro air–liquid interface (ALI) culture of human nasal epithelial cells (hNECs) and used Poly(I:C) as a surrogate of viral components to simulate their effects toward re-epithelization and mucociliary functions of the nasal epithelium following damages from a viral infection.ResultsThrough previous and current viral infection expression data, we found that respiratory viral infection of hNECs downregulated motile cilia gene expression. We then further tested the effects of antiviral response activation on the differentiation of hNECs using Poly(I:C) stimulation on differentiating human nasal epithelial stem/progenitor cells (hNESPCs). Using this model, we observed reduced ciliated cell differentiation compared to goblet cells, reduced protein and mRNA in ciliogenesis-associated markers, and increased mis-assembly and mis-localization of ciliary protein DNAH5 following treatment with 25 μg/ml Poly(I:C) in differentiating hNECs. Additionally, the cilia length and ciliary beat frequency (CBF) were also decreased, which suggest impairment of ciliary function as well.ConclusionOur results suggest that the impairments of ciliogenesis and ciliary function in hNECs may be triggered by specific expression of host antiviral response genes during re-epithelization of the nasal epithelium following viral infection. This event may in turn drive the development and exacerbation of chronic airway inflammatory diseases.

Suppression of influenza virus infection by rhinovirus interference – at the population, individual and cellular levels

“…We attempted to investigate if any of the RV altered host genes would inhibit the replication of IAV. In our transcriptomic analysis, RV and IAV infection commonly upregulated genes related to innate sensing pathways, innate immune cytokine and chemokine signaling pathways, IFNs and antiviral signaling pathways, which agree with previous reports ( Bui et al., 2019 ; Murphy et al., 2020 ; Chen et al., 2020 ). Using a similar dual infection model, Manel Essaidi-Laziosi et al .…”

Transcriptomics of rhinovirus persistence reveals sustained expression of RIG‐I and interferon‐stimulated genes in nasal epithelial cells in vitro

“…Functional pathway analysis revealed cilium function signature in asthmatic airway epithelial cells—out of the 12 downregulated genes, 8 were found to be similarly downregulated at 3 dpi in our study. Chen et al 27 stimulated differentiating hNECs by treatment with poly(I:C) to mimic RNA virus remnants, and found shortened and improper cilia formation in vitro. Similarly, clinical observations of reduced ciliogenesis and mucus hypersecretion have also been reported for chronic upper airway inflammatory diseases such as chronic rhinosinusitis with nasal polyps 17,28 .…”

Moderate Dose Irradiation Induces DNA Damage and Impairments of Barrier and Host Defense in Nasal Epithelial Cells in vitro

“…In our previous study, we found that abnormal ciliogenesis is associated with ultrastructural abnormalities in ciliated cells. 26 Thus, we further investigated the expression of motile cilia assembly markers, such as the outer dynein arm genes and radial spoke head genes. We found that the outer dynein arm heavy chain genes DNAH5, DNAH6, DNAH12 decreased significantly in both IR1 and IR5 groups.…”