title: Is CFTR-delF508 really absent from the apical membrane of the airway epithelium?
abstract: BACKGROUND
  Understanding where mutant CFTR is localised in airway epithelia is essential in guiding the best therapeutic approach to correct the dysfunction of the CFTR protein. The widely held paradigm is that CF patients harbouring the commonest mutation, CFTR-delF508, trap CFTR within the endoplasmic reticulum and target it for degradation. However there are conflicting reports concerning expression and localisation of CFTR-delF508 in lung tissue. To attempt to resolve this fundamental issue we developed a novel approach to measure CFTR-delF508 in the lower airways of patients who have undergone lung transplantation for advanced CF. By sampling CF and non-CF epithelium simultaneously from the same individual, confounding factors of different airway microenvironments which may have influenced previous observations can be overcome.
  
  
  METHODS
  Epithelia sampled by bronchial brushing above (CF) and below (non-CF) the bronchial anastomosis were stained for CFTR and the localisation and level of expression assessed (n = 12).
  
  
  RESULTS
  There was no significant difference in the proportion of tall columnar cells showing CFTR immunostaining as a discrete band at the apical membrane in cells harbouring the CFTR-delF508 mutation compared to non-CF cells (p = 0.21, n = 12). However, the amount of CFTR expressed at the apical surface was reduced by ∼50% in CF cells compared to non-CF cells (p = 0.04, n = 5).
  
  
  CONCLUSIONS
  Our novel observation challenges the prevailing paradigm that CFTR is essentially absent from the apical membrane of respiratory cells harbouring the CFTR-delF508 mutation. Moreover, it raises the possibility that the new generation of CFTR potentiators may offer a realistic therapeutic option for CF patients.
authors.affiliation: Institute of Cellular Medicine, Newcastle University, Newcastle upon Tyne, United Kingdom.
authors.forename: Lee A
authors.initials: LA
authors.lastname: Borthwick
authors.forename: Phil
authors.initials: P
authors.lastname: Botha
authors.forename: Bernard
authors.initials: B
authors.lastname: Verdon
authors.forename: Malcolm J
authors.initials: MJ
authors.lastname: Brodlie
authors.forename: Aaron
authors.initials: A
authors.lastname: Gardner
authors.forename: David
authors.initials: D
authors.lastname: Bourn
authors.forename: Gail E
authors.initials: GE
authors.lastname: Johnson
authors.forename: Mike A
authors.initials: MA
authors.lastname: Gray
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Fisher
mesh_terms: D001980:Bronchi / Q000378:metabolism; D003550:Cystic Fibrosis / Q000235:genetics / Q000378:metabolism*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D006801:Humans; D066298:In Vitro Techniques; D008168:Lung / Q000378:metabolism; D016040:Lung Transplantation; D018613:Microscopy, Confocal; D020545:Respiratory Mucosa / Q000378:metabolism*
chemical_list: D019005:Cystic Fibrosis Transmembrane Conductance Regulator