title: NADPH Oxidase-4 Overexpression Is Associated With Epithelial Ciliary Dysfunction in Neutrophilic Asthma.
abstract: Bronchial epithelial ciliary dysfunction is an important feature of asthma. We sought to determine the role in asthma of neutrophilic inflammation and nicotinamide adenine dinucleotide phosphate (NADPH) oxidases in ciliary dysfunction.
  
  
  
  Bronchial epithelial ciliary function was assessed by using video microscopy in fresh ex vivo epithelial strips from patients with asthma stratified according to their sputum cell differentials and in culture specimens from healthy control subjects and patients with asthma. Bronchial epithelial oxidative damage was determined by 8-oxo-dG expression. Nicotinamide adenine dinucleotide phosphate oxidase (NOX)/dual oxidase (DUOX) expression was assessed in bronchial epithelial cells by using microarrays, with NOX4 and DUOX1/2 expression assessed in bronchial biopsy specimens. Ciliary dysfunction following NADPH oxidase inhibition, using GKT137831, was evaluated in fresh epithelial strips from patients with asthma and a murine model of ovalbumin sensitization and challenge.
  
  
  
  Ciliary beat frequency was impaired in patients with asthma with sputum neutrophilia (n = 11) vs those without (n = 10) (5.8 [0.6] Hz vs 8.8 [0.5] Hz; P = .003) and was correlated with sputum neutrophil count (r = -0.70; P < .001). Primary bronchial epithelial cells expressed DUOX1/2 and NOX4. Levels of 8-oxo-dG and NOX4 were elevated in patients with neutrophilic vs nonneutrophilic asthma, DUOX1 was elevated in both, and DUOX2 was elevated in nonneutrophilic asthma in vivo. In primary epithelial cultures, ciliary dysfunction did not persist, although NOX4 expression and reactive oxygen species generation was increased from patients with neutrophilic asthma. GKT137831 both improved ciliary function in ex vivo epithelial strips (n = 13), relative to the intensity of neutrophilic inflammation, and abolished ciliary dysfunction in the murine asthma model with no reduction in inflammation.
  
  
  
  Ciliary dysfunction is increased in neutrophilic asthma associated with increased NOX4 expression and is attenuated by NADPH oxidase inhibition.
authors.affiliation: Institute for Lung Health, Department of Infection, Immunity & Inflammation, Glenfield Hospital, University of Leicester, Leicester.
authors.forename: Wing-Yan Heidi
authors.initials: WY
authors.lastname: Wan
authors.affiliation: Institute for Lung Health, Department of Infection, Immunity & Inflammation, Glenfield Hospital, University of Leicester, Leicester.
authors.forename: Fay
authors.initials: F
authors.lastname: Hollins
authors.affiliation: Department of Infection, Immunity and Inflammation, University of Leicester, Leicester.
authors.forename: Louise
authors.initials: L
authors.lastname: Haste
authors.affiliation: Institute for Lung Health, Department of Infection, Immunity & Inflammation, Glenfield Hospital, University of Leicester, Leicester.
authors.forename: Lucy
authors.initials: L
authors.lastname: Woodman
authors.affiliation: Centre for PCD Diagnosis and Research, Department of Infection, Immunity and Inflammation, RK Clinical Sciences Building, University of Leicester, Leicester.
authors.forename: Robert A
authors.initials: RA
authors.lastname: Hirst
authors.affiliation: Institute for Lung Health, Department of Infection, Immunity & Inflammation, Glenfield Hospital, University of Leicester, Leicester.
authors.forename: Sarah
authors.initials: S
authors.lastname: Bolton
authors.affiliation: Institute for Lung Health, Department of Infection, Immunity & Inflammation, Glenfield Hospital, University of Leicester, Leicester.
authors.forename: Edith
authors.initials: E
authors.lastname: Gomez
authors.affiliation: Institute for Lung Health, Department of Infection, Immunity & Inflammation, Glenfield Hospital, University of Leicester, Leicester.
authors.forename: Amanda
authors.initials: A
authors.lastname: Sutcliffe
authors.affiliation: Institute for Lung Health, Department of Infection, Immunity & Inflammation, Glenfield Hospital, University of Leicester, Leicester.
authors.forename: Dhananjay
authors.initials: D
authors.lastname: Desai
authors.affiliation: Institute for Lung Health, Department of Infection, Immunity & Inflammation, Glenfield Hospital, University of Leicester, Leicester.
authors.forename: Latifa
authors.initials: L
authors.lastname: Chachi
authors.affiliation: Institute for Lung Health, Department of Infection, Immunity & Inflammation, Glenfield Hospital, University of Leicester, Leicester.
authors.forename: Vijay
authors.initials: V
authors.lastname: Mistry
authors.affiliation: Genkyotex, Geneva, Switzerland.
authors.forename: Cédric
authors.initials: C
authors.lastname: Szyndralewiez
authors.affiliation: Institute for Lung Health, Department of Infection, Immunity & Inflammation, Glenfield Hospital, University of Leicester, Leicester.
authors.forename: Andrew
authors.initials: A
authors.lastname: Wardlaw
authors.affiliation: Institute for Lung Health, Department of Infection, Immunity & Inflammation, Glenfield Hospital, University of Leicester, Leicester.
authors.forename: Ruth
authors.initials: R
authors.lastname: Saunders
authors.affiliation: Department of Infection, Immunity and Inflammation, University of Leicester, Leicester.
authors.forename: Christopher
authors.initials: C
authors.lastname: O'Callaghan
authors.affiliation: Department of Infection, Immunity and Inflammation, University of Leicester, Leicester.
authors.forename: Peter W
authors.initials: PW
authors.lastname: Andrew
authors.affiliation: Institute for Lung Health, Department of Infection, Immunity & Inflammation, Glenfield Hospital, University of Leicester, Leicester. Electronic address: ceb17@le.ac.uk.
authors.forename: Christopher E
authors.initials: CE
authors.lastname: Brightling
mesh_terms: D000328:Adult; D000818:Animals; D001249:Asthma / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology; D002923:Cilia / Q000378:metabolism*; D000074623:Dual Oxidases; D005260:Female; D006801:Humans; D007249:Inflammation / Q000378:metabolism; D008297:Male; D051379:Mice; D008875:Middle Aged; D000074663:NADPH Oxidase 4; D019255:NADPH Oxidases / Q000378:metabolism*; D009504:Neutrophils; D018384:Oxidative Stress; D020545:Respiratory Mucosa / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology; D013223:Statistics as Topic
chemical_list: D000074623:Dual Oxidases; D000074663:NADPH Oxidase 4; D019255:NADPH Oxidases; C498910:NOX4 protein, human; C431513:DUOX1 protein, human; C431515:DUOX2 protein, human
keywords: NOX4; asthma; epithelial cells; oxidative stress
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