Enhanced Ca2+ entry due to Orai1 plasma membrane insertion increases IL-8 secretion by cystic fibrosis airways

Haouaria Balghi, Renaud Robert, Benjamin Rappaz, Xuexin Zhang, Adeline Wohlhuter-Haddad, Alexandra Evagelidis, Yishan Luo, Julie Goepp, Pasquale Ferraro, Philippe Roméo, Mohamed Trebak, Paul W. Wiseman, David Y. Thomas, John W. Hanrahan

Research output: Contribution to journalArticlepeer-review

45 Scopus citations

Abstract

Cystic fibrosis (CF) is caused by mutations in the gene encoding the CF transmembrane conductance regulator (CFTR). The most common mutation, ΔF508, causes retention of CFTR in the endoplasmic reticulum (ER). Some CF abnormalities can be explained by altered Ca2+ homeostasis, although it remains unknown how CFTR influences calcium signaling. This study examined the novel hypothesis that store-operated calcium entry (SOCE) through Orai1 is abnormal in CF. The significance of Orai1- mediated SOCE for increased interleukin-8 (IL-8) expression in CF was also investigated. CF and non-CF human airway epithelial cell line and primary cells (obtained at lung transplantation) were used in Ca2+ imaging, electrophysiology, and fluorescence imaging experiments to explore differences in Orai1 function in CF vs. non-CF cells. Protein expression and localization was assessed by Western blots, cell surface biotinylation, ELISA, and image correlation spectroscopy (ICS). We show here that store-operated Ca2+ entry (SOCE) is elevated in CF human airway epithelial cells (hAECs; ∼1.8- and ∼2.5-fold for total Ca2+ i increase and Ca2+ influx rate, respectively, and ∼2-fold increase in the ICRAC current) and is caused by increased exocytotic insertion (∼2-fold) of Orai1 channels into the plasma membrane, which is normalized by rescue of ΔF508-CFTR trafficking to the cell surface. Augmented SOCE in CF cells is a major factor leading to increased IL-8 secretion (∼2-fold). CFTR normally down-regulates the Orai1/stromal interaction molecule 1 (STIM1) complex, and loss of this inhibition due to the absence of CFTR at the plasma membrane helps to explain the potentiated inflammatory response in CF cells.

Original languageEnglish (US)
Pages (from-to)4274-4291
Number of pages18
JournalFASEB Journal
Volume25
Issue number12
DOIs
StatePublished - Dec 2011

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Genetics

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