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Reduced histone deacetylase 7 activity restores function to misfolded CFTR in cystic fibrosis

Nature Chemical Biology volume 6, pages 25–33 (2010)Cite this article

Abstract

Chemical modulation of histone deacetylase (HDAC) activity by HDAC inhibitors (HDACi) is an increasingly important approach for modifying the etiology of human disease. Loss-of-function diseases arise as a consequence of protein misfolding and degradation, which lead to system failures. The ΔF508 mutation in cystic fibrosis transmembrane conductance regulator (CFTR) results in the absence of the cell surface chloride channel and a loss of airway hydration, leading to the premature lung failure and reduced lifespan responsible for cystic fibrosis. We now show that the HDACi suberoylanilide hydroxamic acid (SAHA) restores surface channel activity in human primary airway epithelia to levels that are 28% of those of wild-type CFTR. Biological silencing of all known class I and II HDACs reveals that HDAC7 plays a central role in restoration of ΔF508 function. We suggest that the tunable capacity of HDACs can be manipulated by chemical biology to counter the onset of cystic fibrosis and other human misfolding disorders.

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Figure 1: HDAC inhibitor treatment increases ΔF508 CFTR expression and trafficking.
Figure 2: Chronic low-dose SAHA treatment increases ΔF508 CFTR stability and trafficking.
Figure 3: HDAC inhibitor treatment activates ΔF508 channel activity.
Figure 4: Silencing of HDAC7 increases ΔF508 CFTR expression, trafficking and activity.
Figure 5: SAHA rescues ΔF508 CFTR activity in primary human bronchial epithelial cells.
Figure 6: Mechanism of HDAC7-mediated ΔF508 multitarget pathway correction.

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Acknowledgements

We thank J.P. Clancy (University of Alabama, Birmingham) for the CFBE41o- expressing ΔF508 and the corrected CFBE41o- (WT-HBE41o-) expressing wild-type CFTR. This work was supported by US National Institutes of Health (NIH) grants HL79442 and GM42336 and aid from the Cystic Fibrosis Consortium to W.E.B.; NIH grant NS055781 to J.M.G.; NIH grants DK68196 and DK72506 and aid from the Cystic Fibrosis Consortium to R.A.F.; NIH grant UR98647 and aid from the Cystic Fibrosis Consortium to E.J.S.; NIH grant AG03197 to J.W.K.; NIH grant DK23567 to J.R.R.; NIH grant AG78594 to G.M.; and NIH grant DK075302 and aid from the Canadian Institutes of Health Research to G.L.L. D.M.H. has received support by fellowships from the Canadian Cystic Fibrosis Foundation and the Canadian Institutes of Health Research; D.H. was supported by a fellowship from the Friedreich's Ataxia Research Alliance. T.O. was supported by a fellowship from the Canadian Cystic Fibrosis Foundation.

Author information

Author notes

  1. Darren M Hutt and David Herman: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Cell Biology, The Scripps Research Institute, La Jolla, California, USA

    Darren M Hutt, Jeanne Matteson, Wendy Kellner, John R Yates III & William E Balch

  2. Department of Molecular Biology, The Scripps Research Institute, La Jolla, California, USA

    David Herman, Ben Hoch, Joel M Gottesfeld & William E Balch

  3. Razavi Newman Center for Bioinformatics, Salk Institute for Biological Studies, La Jolla, California, USA

    Ana P C Rodrigues & Gerard Manning

  4. Department of Cell Biology and Physiology, Allergy and Critical Care Medicine, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA

    Sabrina Noel & Raymond A Frizzell

  5. Division of Pulmonary, Allergy and Critical Care Medicine, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA

    Joseph M Pilewski

  6. Department of Chemistry, The Scripps Research Institute, La Jolla, California, USA

    Jeffery W Kelly

  7. Skaggs Institute of Chemical Biology, The Scripps Research Institute, La Jolla, California, USA

    Jeffery W Kelly

  8. Molecular Biophysics, University of Texas Southwestern Medical Center, Dallas, Texas, USA

    Andre Schmidt & Philip J Thomas

  9. Department of Biochemistry and Molecular Biology, Oregon Health and Sciences University, Portland, Oregon, USA

    Yoshihiro Matsumura & William R Skach

  10. Department of Cell and Developmental Biology, University of North Carolina, Chapel Hill, North Carolina, USA

    Martina Gentzsch

  11. Department of Biochemistry and Biophysics, University of North Carolina, Chapel Hill, North Carolina, USA

    John R Riordan

  12. Department of Cell Biology and Physiology, University of Alabama at Birmingham, Birmingham, Alabama, USA

    Eric J Sorscher

  13. Department of Physiology, McGill University, Montreal, Quebec, Canada

    Tsukasa Okiyoneda & Gergely L Lukacs

  14. Department of Chemical Physiology, The Scripps Research Institute, La Jolla, California, USA

    John R Yates III & William E Balch

  15. The Institute for Childhood and Neglected Diseases, The Scripps Research Institute, La Jolla, California, USA

    William E Balch

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Contributions

The manuscript was written by D.M.H. and W.E.B. D.M.H. from the W.E.B. laboratory was responsible for all western blot assays, iodide efflux data and analyses in CFBE41o- cells. W.K. assisted D.M.H. from the Balch laboratory in providing technical support for assays. D.H. from the J.M.G. laboratory performed all qRT-PCR analysis of CFTR and HDAC expression. A.P.C.R. from the G.M. laboratory performed the microarray analysis. S.N. from the R.A.F. laboratory performed all short-circuit current and western analyses in primary DF/DF-HBE cells, which were obtained through a collaboration with J.M.P. B.H. from the W.E.B. laboratory performed the ChIP analysis. A.S. from the P.J.T. laboratory performed the thermal melt curve analysis of CFTR-NBD1. Y.M. from the W.R.S. laboratory performed the in vitro translation analysis. M.G. performed the cell surface expression of DF508-CFTRextope in primary HBE cells. T.O. from the G.L.L. laboratory performed the cell surface stability analysis. E.J.S. performed the short-circuit analysis of TSA in CFBE41o- cells. J.R.R. performed the cell surface analysis. J.M. from the W.E.B. laboratory performed all pulse chase analyses. J.W.K. laboratory tested the effect of SAHA on L444P-glucoceribrosidase. W.E.B. directed all aspects of the project. R.A.F., P.J.T., E.J.S., W.R.S., J.R.R., M.G., J.M.G., G.L.L., J.R.Y. and G.M. supervised specific aspects of the project. W.E.B., R.A.F., P.J.T., E.J.S., W.R.S., J.R.R., M.G., J.M.G., G.L.L., J.R.Y. and G.M. provided financial support.

Corresponding author

Correspondence to William E Balch.

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Competing interests

J.W.K. is a founder and W.E.B. is a scientific advisor for Proteostasis Therapeutics Inc. of Cambridge, Massachusetts, USA.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–11, Supplementary Table 1 and Supplementary Methods (PDF 8171 kb)

Supplementary Table 2

Microarray data for CFBE41o- cells following HDAC1 and 7 silencing. (XLS 4843 kb)

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Hutt, D., Herman, D., Rodrigues, A. et al. Reduced histone deacetylase 7 activity restores function to misfolded CFTR in cystic fibrosis. Nat Chem Biol 6, 25–33 (2010). https://doi.org/10.1038/nchembio.275

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