Agonist binding to β-adrenergic receptors on human airway epithelial cells inhibits migration and wound repair

Elizabeth R. Peitzman, Nathan A. Zaidman, Peter J. Maniak, Scott M. O’Grady

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Human airway epithelial cells express β-adrenergic receptors (β-ARs), which regulate mucociliary clearance by stimulating transepithelial anion transport and ciliary beat frequency. Previous studies using airway epithelial cells showed that stimulation with isoproterenol increased cell migration and wound repair by a cAMP-dependent mechanism. In the present study, impedance-sensing arrays were used to measure cell migration and epithelial restitution following wounding of confluent normal human bronchial epithelial (NHBE) and Calu-3 cells by electroporation. Stimulation with epinephrine or the β2-AR-selective agonist salbutamol significantly delayed wound closure and reduced the mean surface area of lamellipodia protruding into the wound. Treatment with the β-AR bias agonist carvedilol or isoetharine also produced a delay in epithelial restitution similar in magnitude to epinephrine and salbutamol. Measurements of extracellular signal-regulated kinase phosphorylation following salbutamol or carvedilol stimulation showed no significant change in the level of phosphorylation compared with untreated control cells. However, inhibition of protein phosphatase 2A activity completely blocked the delay in wound closure produced by β-AR agonists. In Calu-3 cells, where CFTR expression was inhibited by RNAi, salbutamol did not inhibit wound repair, suggesting that β-AR agonist stimulation and loss of CFTR function share a common pathway leading to inhibition of epithelial repair. Confocal images of the basal membrane of Calu-3 cells labeled with anti- β1-integrin (clone HUTS-4) antibody showed that treatment with epinephrine or carvedilol reduced the level of activated integrin in the membrane. These findings suggest that treatment with β-AR agonists delays airway epithelial repair by a G protein- and cAMPindependent mechanism involving protein phosphatase 2A and a reduction in β1-integrin activation in the basal membrane.

Original languageEnglish (US)
Pages (from-to)C847-C855
JournalAmerican Journal of Physiology - Cell Physiology
Volume309
Issue number12
DOIs
StatePublished - Dec 15 2015

Fingerprint

Adrenergic Receptors
Albuterol
Cell Movement
Epithelial Cells
Integrins
Epinephrine
Wounds and Injuries
Protein Phosphatase 2
Isoetharine
Membranes
Phosphorylation
Mucociliary Clearance
Pseudopodia
Electroporation
Extracellular Signal-Regulated MAP Kinases
RNA Interference
Electric Impedance
Isoproterenol
GTP-Binding Proteins
Anions

Keywords

  • Biased agonism
  • Carvedilol
  • Epinephrine
  • Impedance sensing
  • β-arrestin signaling

Cite this

Agonist binding to β-adrenergic receptors on human airway epithelial cells inhibits migration and wound repair. / Peitzman, Elizabeth R.; Zaidman, Nathan A.; Maniak, Peter J.; O’Grady, Scott M.

In: American Journal of Physiology - Cell Physiology, Vol. 309, No. 12, 15.12.2015, p. C847-C855.

Research output: Contribution to journalArticle

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