Hypoxia-induced cystic fibrosis transmembrane conductance regulator dysfunction is a universal mechanism underlying reduced mucociliary transport in sinusitis

• Published in: International forum of allergy & rhinology
• Main Authors: Cho, Do-Yeon, Zhang, Shaoyan, Norwood, T Graham, Skinner, Daniel, Hollis, Tonja A, Ehrhardt, Marie L, Yang, Lydia C, Lim, Dong-Jin, Grayson, Jessica W, Lazrak, Ahmed, Matalon, Sadis, Rowe, Steven M, Woodworth, Bradford A
• Format: Journal Article
• Language: English
• Published: United States 01.06.2024
• Subjects: hypoxia; functional microanatomy; patch clamp; rabbit sinusitis; optical coherence tomography; glutathione; mucociliary transport; potential difference; periciliary liquid; chronic rhinosinusitis; sinusitis; airway surface liquid; CFTR; chronic sinusitis
• ISSN: 2042-6984
• DOI: 10.1002/alr.23309

Hypoxia due to sinus obstruction is a major pathogenic mechanism leading to sinusitis. The objective of the current study is to define the electrophysiologic characteristics of hypoxia in vitro and in vivo. Cystic fibrosis bronchoepithelial cells expressing wild-type cystic fibrosis transmembrane conductance regulator (CFTR) and human sinonasal epithelial cells were exposed to 1% or atmospheric O for 24 h. Time-dependent production of cytoplasmic free radicals was measured. Cells were subjected to Ussing chamber and patch clamp technique where CFTR currents were recorded in whole-cell and cell-attached mode for single channel studies. Indices of mucociliary transport (MCT) were measured using micro-optical coherence tomography. In a rabbit hypoxic maxillary sinus model, tissue oxygenation, relative mRNA expression of HIF-1α, pH, sinus potential difference (SPD), and MCT were determined. Ussing chamber (p < 0.05), whole-cell (p < 0.001), and single channel patch-clamp (p < 0.0001) showed significant inhibition of Cl currents in hypoxic cells. Cytoplasmic free radicals showed time-dependent elevation peaking at 4 h (p < 0.0001). Airway surface liquid (p < 0.0001), periciliary liquid (p < 0.001), and MCT (p < 0.01) were diminished. Co-incubation with the free radical scavenger glutathione negated the impact of hypoxia on single channel currents and MCT markers. In sinusitis rabbits, mucosa exhibited low tissue oxygenation (p < 0.0001), increased HIF1α mRNA (p < 0.05), reduced pH (p < 0.01), and decreased MCT (p < 0.001). SPD measurements demonstrated markedly diminished transepithelial Cl transport (p < 0.0001). Hypoxia induces severe CFTR dysfunction via free radical production causing reduced MCT in vitro and in vivo. Improved oxygenation is critical to reducing the impact of persistent mucociliary dysfunction.