Measuring the Effect of Histone Deacetylase Inhibitors (HDACi) on the Secretion and Activity of Alpha-1 Antitrypsin

  • Chao Wang
  • Marion Bouchecareilh
  • William E. BalchEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1639)


Alpha-1 antitrypsin deficiency (AATD) is a protein conformational disease with the most common cause being the Z-variant mutation in alpha-1 antitrypsin (Z-AAT). The misfolded conformation triggered by the Z-variant disrupts cellular proteostasis (protein folding) systems and fails to meet the endoplasmic reticulum (ER) export metrics, leading to decreased circulating AAT and deficient antiprotease activity in the plasma and lung. Here, we describe the methods for measuring the secretion and neutrophil elastase (NE) inhibition activity of AAT/Z-AAT, as well as the response to histone deacetylase inhibitor (HDACi), a major proteostasis modifier that impacts the secretion and function of AATD from the liver to plasma. These methods provide a platform for further therapeutic development of proteostasis regulators for AATD.

Key words

Alpha-1 antitrypsin deficiency Cellular proteostasis Histone deacetylase inhibitor Proteostasis regulators 



This work was supported by grants from National Institute of Health HL095524 for WEB. CW is supported by a postdoctoral research fellowship from Alpha-1 Foundation.


  1. 1.
    Ghouse R, Chu A, Wang Y, Perlmutter DH (2014) Mysteries of alpha1-antitrypsin deficiency: emerging therapeutic strategies for a challenging disease. Dis Model Mech 7:411–419CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Balch WE, Morimoto RI, Dillin A, Kelly JW (2008) Adapting proteostasis for disease intervention. Science 319:916–919CrossRefPubMedGoogle Scholar
  3. 3.
    Bouchecareilh M, Conkright JJ, Balch WE (2010) Proteostasis strategies for restoring alpha1-antitrypsin deficiency. Proc Am Thorac Soc 7:415–422CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Bouchecareilh M, Balch WE (2011) Proteostasis: a new therapeutic paradigm for pulmonary disease. Proc Am Thorac Soc 8:189–195CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Bouchecareilh M, Balch WE (2012) Proteostasis, an emerging therapeutic paradigm for managing inflammatory airway stress disease. Curr Mol Med 12:815–826CrossRefPubMedGoogle Scholar
  6. 6.
    Powers ET, Balch WE (2013) Diversity in the origins of proteostasis networks–a driver for protein function in evolution. Nat Rev Mol Cell Biol 14:237–248CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Gooptu B, Dickens JA, Lomas DA (2014) The molecular and cellular pathology of alpha(1)-antitrypsin deficiency. Trends Mol Med 20:116–127CrossRefPubMedGoogle Scholar
  8. 8.
    Teckman JH (2013) Liver disease in alpha-1 antitrypsin deficiency: current understanding and future therapy. COPD 10(Suppl 1):35–43CrossRefPubMedGoogle Scholar
  9. 9.
    Brebner JA, Stockley RA (2013) Recent advances in alpha-1-antitrypsin deficiency-related lung disease. Expert Rev Respir Med 7:213–229; quiz 230CrossRefPubMedGoogle Scholar
  10. 10.
    Kopito RR, Ron D (2000) Conformational disease. Nat Cell Biol 2:E207–E209CrossRefPubMedGoogle Scholar
  11. 11.
    Nyon MP, Gooptu B (2014) Therapeutic targeting of misfolding and conformational change in alpha1-antitrypsin deficiency. Future Med Chem 6:1047–1065CrossRefPubMedGoogle Scholar
  12. 12.
    Wang C, Balch WE (2015) Managing the adaptive proteostatic landscape: restoring resilience in alpha-1 antitrypsin deficiency. In: Sandhaus RA, Wanner A (eds) Alpha-1 antitrypsin: role in health and disease. Springer, Cham, Switzerland. (in press)Google Scholar
  13. 13.
    Hutt DM, Balch WE (2013) Expanding proteostasis by membrane trafficking networks. Cold Spring Harb Perspect Med 3:1–21Google Scholar
  14. 14.
    Kovacs JJ, Murphy PJ, Gaillard S, Zhao X, Wu JT, Nicchitta CV, Yoshida M, Toft DO, Pratt WB, Yao TP (2005) HDAC6 regulates Hsp90 acetylation and chaperone-dependent activation of glucocorticoid receptor. Mol Cell 18:601–607CrossRefPubMedGoogle Scholar
  15. 15.
    Marinova Z, Ren M, Wendland JR, Leng Y, Liang MH, Yasuda S, Leeds P, Chuang DM (2009) Valproic acid induces functional heat-shock protein 70 via Class I histone deacetylase inhibition in cortical neurons: a potential role of Sp1 acetylation. J Neurochem 111:976–987CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Hageman J, Rujano MA, van Waarde MA, Kakkar V, Dirks RP, Govorukhina N, Oosterveld-Hut HM, Lubsen NH, Kampinga HH (2010) A DNAJB chaperone subfamily with HDAC-dependent activities suppresses toxic protein aggregation. Mol Cell 37:355–369CrossRefPubMedGoogle Scholar
  17. 17.
    Rao R, Nalluri S, Kolhe R, Yang Y, Fiskus W, Chen J, Ha K, Buckley KM, Balusu R, Coothankandaswamy V, Joshi A, Atadja P, Bhalla KN (2010) Treatment with panobinostat induces glucose-regulated protein 78 acetylation and endoplasmic reticulum stress in breast cancer cells. Mol Cancer Ther 9:942–952CrossRefPubMedGoogle Scholar
  18. 18.
    Westerheide SD, Anckar J, Stevens SM Jr, Sistonen L, Morimoto RI (2009) Stress-inducible regulation of heat shock factor 1 by the deacetylase SIRT1. Science 323:1063–1066CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Wagner SA, Beli P, Weinert BT, Nielsen ML, Cox J, Mann M, Choudhary C (2011) A proteome-wide, quantitative survey of in vivo ubiquitylation sites reveals widespread regulatory roles. Mol Cell Proteomics 10:M111 013284CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Janke C, Bulinski JC (2011) Post-translational regulation of the microtubule cytoskeleton: mechanisms and functions. Nat Rev Mol Cell Biol 12:773–786CrossRefPubMedGoogle Scholar
  21. 21.
    Hutt DM, Herman D, Rodrigues AP, Noel S, Pilewski JM, Matteson J, Hoch B, Kellner W, Kelly JW, Schmidt A, Thomas PJ, Matsumura Y, Skach WR, Gentzsch M, Riordan JR, Sorscher EJ, Okiyoneda T, Yates JR 3rd, Lukacs GL, Frizzell RA, Manning G, Gottesfeld JM, Balch WE (2010) Reduced histone deacetylase 7 activity restores function to misfolded CFTR in cystic fibrosis. Nat Chem Biol 6:25–33CrossRefPubMedGoogle Scholar
  22. 22.
    Lu J, Yang C, Chen M, Ye DY, Lonser RR, Brady RO, Zhuang Z (2011) Histone deacetylase inhibitors prevent the degradation and restore the activity of glucocerebrosidase in Gaucher disease. Proc Natl Acad Sci U S A 108:21200–21205CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Moresi V, Williams AH, Meadows E, Flynn JM, Potthoff MJ, McAnally J, Shelton JM, Backs J, Klein WH, Richardson JA, Bassel-Duby R, Olson EN (2010) Myogenin and class II HDACs control neurogenic muscle atrophy by inducing E3 ubiquitin ligases. Cell 143:35–45CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Pipalia NH, Cosner CC, Huang A, Chatterjee A, Bourbon P, Farley N, Helquist P, Wiest O, Maxfield FR (2011) Histone deacetylase inhibitor treatment dramatically reduces cholesterol accumulation in Niemann-Pick type C1 mutant human fibroblasts. Proc Natl Acad Sci U S A 108:5620–5625CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Coppede F (2014) The potential of epigenetic therapies in neurodegenerative diseases. Front Genet 5:220PubMedPubMedCentralGoogle Scholar
  26. 26.
    Bouchecareilh M, Hutt DM, Szajner P, Flotte TR, Balch WE (2012) Histone deacetylase inhibitor (HDACi) suberoylanilide hydroxamic acid (SAHA)-mediated correction of alpha1-antitrypsin deficiency. J Biol Chem 287:38265–38278CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Hebert DN, Lamriben L, Powers ET, Kelly JW (2014) The intrinsic and extrinsic effects of N-linked glycans on glycoproteostasis. Nat Chem Biol 10:902–910CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Gettins PG (2002) Serpin structure, mechanism, and function. Chem Rev 102:4751–4804CrossRefPubMedGoogle Scholar
  29. 29.
    Hidvegi T, Ewing M, Hale P, Dippold C, Beckett C, Kemp C, Maurice N, Mukherjee A, Goldbach C, Watkins S, Michalopoulos G, Perlmutter DH (2010) An autophagy-enhancing drug promotes degradation of mutant alpha1-antitrypsin Z and reduces hepatic fibrosis. Science 329:229–232CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Chao Wang
    • 1
  • Marion Bouchecareilh
    • 2
  • William E. Balch
    • 1
    Email author
  1. 1.Department of Molecular Medicine, The Skaggs Institute for Chemical BiologyThe Scripps Research InstituteLa JollaUSA
  2. 2.Institut de Biochimie et Génétique Cellulaires, CNRS UMR 5095Université de BordeauxBordeauxFrance

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