LPS decreases CFTR open probability and mucociliary transport through generation of reactive oxygen species

Do Yeon Cho; Shaoyan Zhang; Ahmed Lazrak; Daniel Skinner; Harrison M Thompson; Jessica Grayson; Purushotham Guroji; Saurabh Aggarwal; Zsuzsanna Bebok; Steven M Rowe; Sadis Matalon; Eric J Sorscher; Bradford A Woodworth

doi:10.1016/j.redox.2021.101998

LPS decreases CFTR open probability and mucociliary transport through generation of reactive oxygen species

Redox Biol. 2021 Jul:43:101998. doi: 10.1016/j.redox.2021.101998. Epub 2021 Apr 30.

Authors

Affiliations

¹ Department of Otolaryngology Head & Neck Surgery, University of Alabama at Birmingham, Birmingham, AL, USA; Gregory Fleming James Cystic Fibrosis Research Center, University of Alabama at Birmingham, Birmingham, AL, USA; Division of Otolaryngology, Department of Surgery, Veterans Affairs, Birmingham, AL, USA.
² Department of Otolaryngology Head & Neck Surgery, University of Alabama at Birmingham, Birmingham, AL, USA; Gregory Fleming James Cystic Fibrosis Research Center, University of Alabama at Birmingham, Birmingham, AL, USA.
³ Gregory Fleming James Cystic Fibrosis Research Center, University of Alabama at Birmingham, Birmingham, AL, USA; Department of Anesthesiology, University of Alabama at Birmingham, Birmingham, AL, USA.
⁴ Department of Otolaryngology Head & Neck Surgery, University of Alabama at Birmingham, Birmingham, AL, USA.
⁵ Department of Cell Developmental and Integrative Biology, University of Alabama at Birmingham, Birmingham, AL, USA.
⁶ Gregory Fleming James Cystic Fibrosis Research Center, University of Alabama at Birmingham, Birmingham, AL, USA; Department of Cell Developmental and Integrative Biology, University of Alabama at Birmingham, Birmingham, AL, USA.
⁷ Gregory Fleming James Cystic Fibrosis Research Center, University of Alabama at Birmingham, Birmingham, AL, USA; Department of Cell Developmental and Integrative Biology, University of Alabama at Birmingham, Birmingham, AL, USA; Department of Pediatrics, University of Alabama at Birmingham, Birmingham, AL, USA; Department of Medicine, University of Alabama at Birmingham, Birmingham, AL, USA.
⁸ Department of Pediatrics, Emory University, Atlanta, GA, USA.
⁹ Department of Otolaryngology Head & Neck Surgery, University of Alabama at Birmingham, Birmingham, AL, USA; Gregory Fleming James Cystic Fibrosis Research Center, University of Alabama at Birmingham, Birmingham, AL, USA. Electronic address: bwoodworth@uabmc.edu.

Abstract

Lipopolysaccharide (LPS) serves as the interface between gram-negative bacteria (GNB) and the innate immune response in respiratory epithelial cells (REC). Herein, we describe a novel biological role of LPS that permits GNB to persist in the respiratory tract through inducing CFTR and mucociliary dysfunction. LPS reduced cystic fibrosis transmembrane conductance regulater (CFTR)-mediated short-circuit current in mammalian REC in Ussing chambers and nearly abrogated CFTR single channel activity (defined as forskolin-activated Cl^- currents) in patch clamp studies, effects of which were blocked with toll-like receptor (TLR)-4 inhibitor. Unitary conductance and single-channel amplitude of CFTR were unaffected, but open probability and number of active channels were markedly decreased. LPS increased cytoplasmic and mitochondrial reactive oxygen species resulting in CFTR carbonylation. All effects of exposure were eliminated when reduced glutathione was added in the medium along with LPS. Functional microanatomy parameters, including mucociliary transport, in human sinonasal epithelial cells in vitro were also decreased, but restored with co-incubation with glutathione or TLR-4 inhibitor. In vivo measurements, following application of LPS in the nasal cavities showed significant decreases in transepithelial Cl^- secretion as measured by nasal potential difference (NPD) - an effect that was nullified with glutathione and TLR-4 inhibitor. These data provide definitive evidence that LPS-generated reactive intermediates downregulate CFTR function in vitro and in vivo which results in cystic fibrosis-type disease. Findings have implications for therapeutic approaches intent on stimulating Cl^- secretion and/or reducing oxidative stress to decrease the sequelae of GNB airway colonization and infection.

Keywords: Chronic rhinosinusitis; Cystic fibrosis transmembrane conductance regulator; Lipopolysaccharide; Micro optical coherence tomography; Mucociliary transport; Oxygen species; Reactive.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Animals
Cystic Fibrosis Transmembrane Conductance Regulator*
Epithelial Cells / metabolism
Humans
Ion Transport
Lipopolysaccharides
Mucociliary Clearance*
Probability
Reactive Oxygen Species

Substances

CFTR protein, human
Lipopolysaccharides
Reactive Oxygen Species
Cystic Fibrosis Transmembrane Conductance Regulator

Abstract

Publication types

MeSH terms

Substances

Grants and funding