Inhibition of ATP hydrolysis restores airway surface liquid production in cystic fibrosis airway epithelia

• Published in: American journal of physiology. Lung cellular and molecular physiology 2020-02, Vol.318 (2), p.L356-L365
• Main Authors: van Heusden, Catharina, Button, Brian, Anderson, Wayne H, Ceppe, Agathe, Morton, Lisa C, O'Neal, Wanda K, Dang, Hong, Alexis, Neil E, Donaldson, Scott, Stephan, Holger, Boucher, Richard C, Lazarowski, Eduardo R
• Format: Article
• Language: English
• Subjects: Adenosine Triphosphatases - genetics; Adenosine Triphosphatases - metabolism; Adenosine Triphosphate - metabolism; Alkaline Phosphatase - genetics; Alkaline Phosphatase - metabolism; Bronchi - pathology; Cystic Fibrosis - metabolism; Cystic Fibrosis - pathology; Enzyme Inhibitors - pharmacology; Epithelial Cells - drug effects; Epithelial Cells - metabolism; Humans; Hydrolysis; Respiratory Mucosa - drug effects; Respiratory Mucosa - metabolism; Respiratory Mucosa - pathology; Sputum - enzymology; Tungsten Compounds - pharmacology
• ISSN: 1040-0605; 1522-1504
• DOI: 10.1152/AJPLUNG.00449.2019

Airway surface dehydration is a pathological feature of cystic fibrosis (CF) lung disease. CF is caused by mutations in the CF transmembrane conductance regulator (CFTR), a cyclic AMP-regulated Cl channel controlled in part by the adenosine A receptor. An alternative CFTR-independent mechanism of fluid secretion is regulated by ATP via the P2Y receptor (P2Y R) that activates Ca -regulated Cl channels (CaCC/TMEM16) and inhibits Na absorption. However, due to rapid ATP hydrolysis, steady-state ATP levels in CF airway surface liquid (ASL) are inadequate to maintain P2Y R-mediated fluid secretion. Therefore, inhibiting airway epithelial ecto-ATPases to increase ASL ATP levels constitutes a strategy to restore airway surface hydration in CF. Using [γ P]ATP as radiotracer, we assessed the effect of a series of ATPase inhibitory compounds on the stability of physiologically occurring ATP concentrations. We identified the polyoxometalate [Co (H O) (PW O ) ] (POM-5) as the most potent and effective ecto-ATPase inhibitor in CF airway epithelial cells. POM-5 caused long-lasting inhibition of ATP hydrolysis in airway epithelia, which was reversible upon removal of the inhibitor. Importantly, POM-5 markedly enhanced steady-state levels of released ATP, promoting increased ASL volume in CF cell surfaces. These results provide proof of concept for ecto-ATPase inhibitors as therapeutic agents to restore hydration of CF airway surfaces. As a test of this notion, cell-free sputum supernatants from CF subjects were studied and found to have abnormally elevated ATPase activity, which was markedly inhibited by POM-5.