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Calcium-pump inhibitors induce functional surface expression of ΔF508-CFTR protein in cystic fibrosis epithelial cells

Nature Medicine volume 8, pages 485–492 (2002)Cite this article

Abstract

The most common mutation in cystic fibrosis, ΔF508, results in a cystic fibrosis transmembrane conductance regulator (CFTR) protein that is retained in the endoplasmic reticulum (ER). Retention is dependent upon chaperone proteins, many of which require Ca++ for optimal activity. Interfering with chaperone activity by depleting ER Ca++ stores might allow functional ΔF508-CFTR to reach the cell surface. We exposed several cystic fibrosis cell lines to the ER Ca++ pump inhibitor thapsigargin and evaluated surface expression of ΔF508-CFTR. Treatment released ER-retained ΔF508-CFTR to the plasma membrane, where it functioned effectively as a Cl− channel. Treatment with aerosolized calcium-pump inhibitors reversed the nasal epithelial potential defect observed in a mouse model of ΔF508-CFTR expression. Thus, ER calcium-pump inhibitors represent a potential target for correcting the cystic fibrosis defect.

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Figure 1: CFTR chloride channel activity in CF-affected epithelial cells in control conditions or after treatment with thapsigargin.
Figure 2: Effects of thapsigargin treatment on plasma membrane and transepithelial Cl− permeability.
Figure 3: Immunolocalization of ΔF508-CFTR in untreated and thapsigargin-treated CF epithelial cells.
Figure 4: Chaperone expression and distribution in thapsigargin-treated cells.
Figure 5: NPD measurements in thapsigargin-treated and untreated ΔF508 mutant mice.

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Acknowledgements

We thank M. Drumm for ΔF508-CFTR mice; W. Guggino, M. Blaustein, P. Aronson, J. Burger, G. Fyfe, G. Giebisch, G. Haddad, P. De Camilli, K. Bottomly, R. Lifton and members of the Caplan lab group for suggestions and readings of the manuscript; S.A. Mentone for help with electron microscopy; V. Rajendran and M.W. Nason for technical support; and M. Kashgarian for assistance in evaluating histopathologic specimens. This work was supported by the Alyward Family/Pitney Bowes Gift Fund, Panacea Pharmaceuticals (M.E.E. and M.J.C.) and by NIH grants DK53428 (to M.E.E.), DK50230 (to J.G.), HD32573 (to J.G.), GM42136 (to M.J.C.) and DK17433 (to J.G. and M.J.C.).

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Author notes

  1. Judith Glöckner-Pagel and Catherine A. Ambrose: J.G.-P. and C.A.A. contributed equally to this study.

Authors and Affiliations

  1. Department of Pediatrics, Yale University School of Medicine, New Haven, Connecticut, USA

    Marie E. Egan, Catherine A. Ambrose, Paula A. Cahill, Lamiko Pappoe & Naomi Balamuth

  2. Department of Surgery, Yale University School of Medicine, New Haven, Connecticut, USA

    John Geibel

  3. Department of Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, Connecticut, USA

    Marie E. Egan, Judith Glöckner-Pagel, Edward Cho, Susan Canny, Carsten A. Wagner, John Geibel & Michael J. Caplan

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Correspondence to Michael J. Caplan.

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Competing interests

These studies were supported to a small extent by a sponsored research agreement between Yale University and Panacea Pharmaceuticals, a small biotech firm that has licensed this technology from Yale University.

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Egan, M., Glöckner-Pagel, J., Ambrose, C. et al. Calcium-pump inhibitors induce functional surface expression of ΔF508-CFTR protein in cystic fibrosis epithelial cells. Nat Med 8, 485–492 (2002). https://doi.org/10.1038/nm0502-485

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