Airway surface hyperviscosity and defective mucociliary transport by IL-17/TNF-α are corrected by β-adrenergic stimulus
Daniela Guidone, … , Isabelle Sermet, Luis J.V. Galietta
Daniela Guidone, … , Isabelle Sermet, Luis J.V. Galietta
Published October 11, 2022
Citation Information: JCI Insight. 2022;7(22):e164944. https://doi.org/10.1172/jci.insight.164944.
View: Text | PDF
Research Article Pulmonology

Airway surface hyperviscosity and defective mucociliary transport by IL-17/TNF-α are corrected by β-adrenergic stimulus

Abstract

The fluid covering the surface of airway epithelia represents a first barrier against pathogens. The chemical and physical properties of the airway surface fluid are controlled by the activity of ion channels and transporters. In cystic fibrosis (CF), loss of CFTR chloride channel function causes airway surface dehydration, bacterial infection, and inflammation. We investigated the effects of IL-17A plus TNF-α, 2 cytokines with relevant roles in CF and other chronic lung diseases. Transcriptome analysis revealed a profound change with upregulation of several genes involved in ion transport, antibacterial defense, and neutrophil recruitment. At the functional level, bronchial epithelia treated in vitro with the cytokine combination showed upregulation of ENaC channel, ATP12A proton pump, ADRB2 β-adrenergic receptor, and SLC26A4 anion exchanger. The overall result of IL-17A/TNF-α treatment was hyperviscosity of the airway surface, as demonstrated by fluorescence recovery after photobleaching (FRAP) experiments. Importantly, stimulation with a β-adrenergic agonist switched airway surface to a low-viscosity state in non-CF but not in CF epithelia. Our study suggests that CF lung disease is sustained by a vicious cycle in which epithelia cannot exit from the hyperviscous state, thus perpetuating the proinflammatory airway surface condition.

Authors

Daniela Guidone, Martina Buccirossi, Paolo Scudieri, Michele Genovese, Sergio Sarnataro, Rossella De Cegli, Federico Cresta, Vito Terlizzi, Gabrielle Planelles, Gilles Crambert, Isabelle Sermet, Luis J.V. Galietta

×

Figure 6

Mechanism of ENaC upregulation by IL-17/TNF-α.

Options: View larger image (or click on image) Download as PowerPoint
Mechanism of ENaC upregulation by IL-17/TNF-α. (A) Representative short-...
(A) Representative short-circuit current traces (left) and summary of data (right) for experiments on epithelia treated with/without IL-17/TNF-α for 72 hours. During recordings, elastase (1.5 μM) and amiloride (10 μM) were sequentially added. Where indicated, epithelia were apically treated with camostat (3 μM) for 18 hours. The scatter dot plot shows, for experiments with camostat pretreatment, the ratio I2/I1, where I1 and I2 are the current amplitudes before and after elastase, respectively (**P < 0.01; Student’s t test). (B) Representative traces from experiments with/without IL-17/TNF-α treatment in which camostat (3 μM) and amiloride were sequentially added. Where indicated, experiments also included addition of elastase (1.5 μM) after camostat. (C) Summary of data showing the rate of ENaC current decay after camostat addition (C, control; I+T, IL-17/TNF-α). Data (t1/2) report the time at which the current decayed to half of initial amplitude (***P < 0.001; Student’s t test). (D) Representative confocal microscope images, from control- and IL-17/TNF-α–treated epithelia, in which SCNN1A, ZO-1, and cilia were detected by immunofluorescence. Scale bar: 25 μm. (E and F) Representative short-circuit current traces and summary of data for experiments where control- and IL-17/TNF-α–treated epithelia were apically exposed to 25 μM GSK650394 (E) or 5 μM PP242 (F) before amiloride. The scatter dot plots report the t1/2 values for each condition. (*P < 0.05 versus control; Student’s t test).