Anchored PDE4 regulates chloride conductance in wild-type and Δf508-CFTR human airway epithelia

Elise Blanchard, Lorna Zlock, Anna Lao, Delphine Mika, Wan Namkung, Moses Xie, Colleen Scheitrum, Dieter C. Gruenert, Alan S. Verkman, Walter E. Finkbeiner, Marco Conti, Wito Richter

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Cystic fibrosis (CF) is caused by mutations in the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR) that impair its expression and/or chloride channel function. Here, we provide evidence that type 4 cyclic nucleotide phosphodiesterases (PDE4s) are critical regulators of the cAMP/PKA-dependent activation of CFTR in primary human bronchial epithelial cells. In non-CF cells, PDE4 inhibition increased CFTR activity under basal conditions (ΔISC 7.1 μA/cm2)) and after isoproterenol stimulation (increased ΔISC from 13.9 to 21.0 μA/cm2) and slowed the return of stimulated CFTR activity to basal levels by >3-fold. In cells homozygous for ΔF508-CFTR, the most common mutation found in CF, PDE4 inhibition alone produced minimal channel activation. However, PDE4 inhibition strongly amplified the effects of CFTR correctors, drugs that increase expression and membrane localization of CFTR, and/or CFTR potentiators, drugs that increase channel gating, to reach ∼25% of the chloride conductance observed in non-CF cells. Biochemical studies indicate that PDE4s are anchored to CFTR and mediate a local regulation of channel function. Taken together, our results implicate PDE4 as an important determinant of CFTR activity in airway epithelia, and support the use of PDE4 inhibitors to potentiate the therapeutic benefits of CFTR correctors and potentiators.

Original languageEnglish
Pages (from-to)791-801
Number of pages11
JournalFASEB Journal
Volume28
Issue number2
DOIs
Publication statusPublished - 2014 Jan 1

Fingerprint

Cystic Fibrosis Transmembrane Conductance Regulator
Chlorides
Epithelium
Cystic Fibrosis
Fibrosis
Chemical activation
Type 4 Cyclic Nucleotide Phosphodiesterase
Phosphodiesterase 4 Inhibitors
Mutation
Chloride Channels
Gene encoding
Isoproterenol
Pharmaceutical Preparations
Epithelial Cells

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Genetics

Cite this

Blanchard, Elise ; Zlock, Lorna ; Lao, Anna ; Mika, Delphine ; Namkung, Wan ; Xie, Moses ; Scheitrum, Colleen ; Gruenert, Dieter C. ; Verkman, Alan S. ; Finkbeiner, Walter E. ; Conti, Marco ; Richter, Wito. / Anchored PDE4 regulates chloride conductance in wild-type and Δf508-CFTR human airway epithelia. In: FASEB Journal. 2014 ; Vol. 28, No. 2. pp. 791-801.
@article{20e75776a1f14e30a5e91a1f32095fd3,
title = "Anchored PDE4 regulates chloride conductance in wild-type and Δf508-CFTR human airway epithelia",
abstract = "Cystic fibrosis (CF) is caused by mutations in the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR) that impair its expression and/or chloride channel function. Here, we provide evidence that type 4 cyclic nucleotide phosphodiesterases (PDE4s) are critical regulators of the cAMP/PKA-dependent activation of CFTR in primary human bronchial epithelial cells. In non-CF cells, PDE4 inhibition increased CFTR activity under basal conditions (ΔISC 7.1 μA/cm2)) and after isoproterenol stimulation (increased ΔISC from 13.9 to 21.0 μA/cm2) and slowed the return of stimulated CFTR activity to basal levels by >3-fold. In cells homozygous for ΔF508-CFTR, the most common mutation found in CF, PDE4 inhibition alone produced minimal channel activation. However, PDE4 inhibition strongly amplified the effects of CFTR correctors, drugs that increase expression and membrane localization of CFTR, and/or CFTR potentiators, drugs that increase channel gating, to reach ∼25{\%} of the chloride conductance observed in non-CF cells. Biochemical studies indicate that PDE4s are anchored to CFTR and mediate a local regulation of channel function. Taken together, our results implicate PDE4 as an important determinant of CFTR activity in airway epithelia, and support the use of PDE4 inhibitors to potentiate the therapeutic benefits of CFTR correctors and potentiators.",
author = "Elise Blanchard and Lorna Zlock and Anna Lao and Delphine Mika and Wan Namkung and Moses Xie and Colleen Scheitrum and Gruenert, {Dieter C.} and Verkman, {Alan S.} and Finkbeiner, {Walter E.} and Marco Conti and Wito Richter",
year = "2014",
month = "1",
day = "1",
doi = "10.1096/fj.13-240861",
language = "English",
volume = "28",
pages = "791--801",
journal = "FASEB Journal",
issn = "0892-6638",
publisher = "FASEB",
number = "2",

}

Blanchard, E, Zlock, L, Lao, A, Mika, D, Namkung, W, Xie, M, Scheitrum, C, Gruenert, DC, Verkman, AS, Finkbeiner, WE, Conti, M & Richter, W 2014, 'Anchored PDE4 regulates chloride conductance in wild-type and Δf508-CFTR human airway epithelia', FASEB Journal, vol. 28, no. 2, pp. 791-801. https://doi.org/10.1096/fj.13-240861

Anchored PDE4 regulates chloride conductance in wild-type and Δf508-CFTR human airway epithelia. / Blanchard, Elise; Zlock, Lorna; Lao, Anna; Mika, Delphine; Namkung, Wan; Xie, Moses; Scheitrum, Colleen; Gruenert, Dieter C.; Verkman, Alan S.; Finkbeiner, Walter E.; Conti, Marco; Richter, Wito.

In: FASEB Journal, Vol. 28, No. 2, 01.01.2014, p. 791-801.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Anchored PDE4 regulates chloride conductance in wild-type and Δf508-CFTR human airway epithelia

AU - Blanchard, Elise

AU - Zlock, Lorna

AU - Lao, Anna

AU - Mika, Delphine

AU - Namkung, Wan

AU - Xie, Moses

AU - Scheitrum, Colleen

AU - Gruenert, Dieter C.

AU - Verkman, Alan S.

AU - Finkbeiner, Walter E.

AU - Conti, Marco

AU - Richter, Wito

PY - 2014/1/1

Y1 - 2014/1/1

N2 - Cystic fibrosis (CF) is caused by mutations in the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR) that impair its expression and/or chloride channel function. Here, we provide evidence that type 4 cyclic nucleotide phosphodiesterases (PDE4s) are critical regulators of the cAMP/PKA-dependent activation of CFTR in primary human bronchial epithelial cells. In non-CF cells, PDE4 inhibition increased CFTR activity under basal conditions (ΔISC 7.1 μA/cm2)) and after isoproterenol stimulation (increased ΔISC from 13.9 to 21.0 μA/cm2) and slowed the return of stimulated CFTR activity to basal levels by >3-fold. In cells homozygous for ΔF508-CFTR, the most common mutation found in CF, PDE4 inhibition alone produced minimal channel activation. However, PDE4 inhibition strongly amplified the effects of CFTR correctors, drugs that increase expression and membrane localization of CFTR, and/or CFTR potentiators, drugs that increase channel gating, to reach ∼25% of the chloride conductance observed in non-CF cells. Biochemical studies indicate that PDE4s are anchored to CFTR and mediate a local regulation of channel function. Taken together, our results implicate PDE4 as an important determinant of CFTR activity in airway epithelia, and support the use of PDE4 inhibitors to potentiate the therapeutic benefits of CFTR correctors and potentiators.

AB - Cystic fibrosis (CF) is caused by mutations in the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR) that impair its expression and/or chloride channel function. Here, we provide evidence that type 4 cyclic nucleotide phosphodiesterases (PDE4s) are critical regulators of the cAMP/PKA-dependent activation of CFTR in primary human bronchial epithelial cells. In non-CF cells, PDE4 inhibition increased CFTR activity under basal conditions (ΔISC 7.1 μA/cm2)) and after isoproterenol stimulation (increased ΔISC from 13.9 to 21.0 μA/cm2) and slowed the return of stimulated CFTR activity to basal levels by >3-fold. In cells homozygous for ΔF508-CFTR, the most common mutation found in CF, PDE4 inhibition alone produced minimal channel activation. However, PDE4 inhibition strongly amplified the effects of CFTR correctors, drugs that increase expression and membrane localization of CFTR, and/or CFTR potentiators, drugs that increase channel gating, to reach ∼25% of the chloride conductance observed in non-CF cells. Biochemical studies indicate that PDE4s are anchored to CFTR and mediate a local regulation of channel function. Taken together, our results implicate PDE4 as an important determinant of CFTR activity in airway epithelia, and support the use of PDE4 inhibitors to potentiate the therapeutic benefits of CFTR correctors and potentiators.

UR - http://www.scopus.com/inward/record.url?scp=84897073798&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84897073798&partnerID=8YFLogxK

U2 - 10.1096/fj.13-240861

DO - 10.1096/fj.13-240861

M3 - Article

VL - 28

SP - 791

EP - 801

JO - FASEB Journal

JF - FASEB Journal

SN - 0892-6638

IS - 2

ER -