Up-regulation of AMP-activated kinase by dysfunctional cystic fibrosis transmembrane conductance regulator in cystic fibrosis airway epithelial cells mitigates excessive inflammation.
AMP-activated kinase (AMPK) is a ubiquitous metabolic sensor that inhibits the cystic fibrosis (CF) transmembrane conductance regulator (CFTR). To determine whether CFTR reciprocally regulates AMPK function in airway epithelia and whether such regulation is involved in lung inflammation, AMPK localization, expression, and activity and cellular metabolic profiles were compared as a function of CFTR status in CF and non-CF primary human bronchial epithelial (HBE) cells. As compared with non-CF HBE cells, CF cells had greater and more diffuse AMPK staining and had greater AMPK activity than their morphologically matched non-CF counterparts. The cellular [AMP]/[ATP] ratio was higher in undifferentiated than in differentiated non-CF cells, which correlated with AMPK activity under these conditions. However, this nucleotide ratio did not predict AMPK activity in differentiating CF cells. Inhibiting channel activity in non-CF cells did not affect AMPK activity or metabolic status, but expressing functional CFTR in CF cells reduced AMPK activity without affecting cellular [AMP]/[ATP]. Therefore, lack of functional CFTR expression and not loss of channel activity in CF cells appears to up-regulate AMPK activity in CF HBE cells, presumably through non-metabolic effects on upstream regulatory pathways. Compared with wild-type CFTR-expressing immortalized CF bronchial epithelial (CFBE) cells, DeltaF508-CFTR-expressing CFBE cells had greater AMPK activity and greater secretion of tumor necrosis factor-alpha and the interleukins IL-6 and IL-8. Further pharmacologic AMPK activation inhibited inflammatory mediator secretion in both wild type- and DeltaF508-expressing cells, suggesting that AMPK activation in CF airway cells is an adaptive response that reduces inflammation. We propose that therapies to activate AMPK in the CF airway may be beneficial in reducing excessive airway inflammation, a major cause of CF morbidity.
Hallows
Kenneth R
KR
Department of Medicine, University of Pittsburgh School of Medicine, Pennsylvania 15261, USA. hallows@pitt.edu
Fitch
Adam C
AC
Richardson
Christine A
CA
Reynolds
Paul R
PR
Clancy
John P
JP
Dagher
Pierre C
PC
Witters
Lee A
LA
Kolls
Jay K
JK
Pilewski
Joseph M
JM
D055372:AMP-Activated Protein Kinases; D000595:Amino Acid Sequence; D001980:Bronchi / Q000201:enzymology*; D002454:Cell Differentiation; D016764:Cell Polarity; D003550:Cystic Fibrosis / Q000201:enzymology*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000502:physiology*; D004847:Epithelial Cells / Q000201:enzymology; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D007249:Inflammation / Q000209:etiology*; D018836:Inflammation Mediators / Q000502:physiology; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000235:genetics* / Q000502:physiology; D017346:Protein Serine-Threonine Kinases / Q000235:genetics* / Q000502:physiology; D015854:Up-Regulation
D018836:Inflammation Mediators; D009097:Multienzyme Complexes; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D017346:Protein Serine-Threonine Kinases; D055372:AMP-Activated Protein Kinases