Autophagy regulates DUOX1 localization and superoxide production in airway epithelial cells during chronic IL-13 stimulation

Autor: Steven L. Brody, Xavier De Deken, Kristi J. Warren, Jenea M. Sweeter, Matthew C. Zimmerman, Iman M. Ahmad, John D. Dickinson
Jazyk: angličtina
Rok vydání: 2017
Předmět:
0301 basic medicine
Autophagosome
IL-33
Interleukin 33
Clinical Biochemistry
Epithelial cells
Biochemistry
chemistry.chemical_compound
0302 clinical medicine
Superoxides
ALI
Air liquid interface
IL-4
Interleukin 4
Electron Paramagnetic Resonance Spectroscopy
lcsh:QH301-705.5
Lung
chemistry.chemical_classification
lcsh:R5-920
Mice
Inbred BALB C
NADPH oxidase
Interleukin-13
biology
Superoxide
CM-H2DCFDA
chloromethyl-2′
7′-dichlorodihydrofluorescein
Dual Oxidases
Cell biology
EGF
Epidermal Growth Factor
030220 oncology & carcinogenesis
IL-13
Interleukin 13
Pneumologie
lcsh:Medicine (General)
NADPH
Nicotinamide adenosine
dinucleotide phosphate
Research Paper
EPR
Electron Paramagnetic Resonance
PVDF
polyvinylidene fluoride
ATG5
CMH
1-hydroxy-3-methoxycarbonyl-2
2
5
5-tetramethylpyrrolidine
hTEC
human tracheobronchial epithelial cells
Cell Line
03 medical and health sciences
ROS
reactive oxygen species
SOD
superoxide dismutase
IL-13
Interleukin 13
Autophagy
Animals
Humans
DUOX1
PBS
Ovalbumin (OVA) phosphate buffered saline
Inflammation
Reactive oxygen species
DUOX1
Dual oxidase 1
Organic Chemistry
IL-4
Apical membrane
BSA
Bovine serum albumen
Asthma
Chimie organique
030104 developmental biology
lcsh:Biology (General)
chemistry
biology.protein
Zdroj: Redox Biology
Redox Biology, 14
Redox Biology, Vol 14, Iss, Pp 272-284 (2018)
ISSN: 2213-2317
Popis: The airway epithelium is a broad interface with the environment, mandating well-orchestrated responses to properly modulate inflammation. Classically, autophagy is a homeostatic pathway triggered in response to external cellular stresses, and is elevated in chronic airway diseases. Recent findings highlight the additional role of autophagy in vesicle trafficking and protein secretion, implicating autophagy pathways in complex cellular responses in disease. Th2 cytokines, IL-13 and IL-4, are increased in asthma and other airway diseases contributing to chronic inflammation. Previously, we observed that IL-13 increases reactive oxygen species (ROS) in airway epithelial cells in an autophagy-dependent fashion. Here, we tested our hypothesis that autophagy is required for IL-13-mediated superoxide production via the NADPH oxidase DUOX1. Using a mouse model of Th2-mediated inflammation induced by OVA-allergen, we observed elevated lung amounts of IL-13 and IL-4 accompanied by increased autophagosome levels, determined by LC3BII protein levels and immunostaining. ROS levels were elevated and DUOX1 expression was increased 70-fold in OVA-challenged lungs. To address the role of autophagy and ROS in the airway epithelium, we treated primary human tracheobronchial epithelial cells with IL-13 or IL-4. Prolonged, 7-day treatment increased autophagosome formation and degradation, while brief activation had no effect. Under parallel culture conditions, IL-13 and IL-4 increased intracellular superoxide levels as determined by electron paramagnetic resonance (EPR) spectroscopy. Prolonged IL-13 activation increased DUOX1, localized at the apical membrane. Silencing DUOX1 by siRNA attenuated IL-13-mediated increases in superoxide, but did not reduce autophagy activities. Notably, depletion of autophagy regulatory protein ATG5 significantly reduced superoxide without diminishing total DUOX1 levels. Depletion of ATG5, however, diminished DUOX1 localization at the apical membrane. The findings suggest non-canonical autophagy activity regulates DUOX1-dependent localization required for intracellular superoxide production during Th2 inflammation. Thus, in chronic Th2 inflammatory airway disease, autophagy proteins may be responsible for persistent intracellular superoxide production.
SCOPUS: ar.j
info:eu-repo/semantics/published
Databáze: OpenAIRE
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