Advertisement

Molecular Medicine

, Volume 19, Issue 1, pp 79–87 | Cite as

Preclinical Studies in the mdx Mouse Model of Duchenne Muscular Dystrophy with the Histone Deacetylase Inhibitor Givinostat

  • Silvia Consalvi
  • Chiara Mozzetta
  • Paolo Bettica
  • Massimiliano Germani
  • Francesco Fiorentini
  • Francesca Del Bene
  • Maurizio Rocchetti
  • Flavio Leoni
  • Valmen Monzani
  • Paolo Mascagni
  • Pier Lorenzo Puri
  • Valentina Saccone
Research Article

Abstract

Previous work has established the existence of dystrophin-nitric oxide (NO) signaling to histone deacetylases (HDACs) that is deregulated in dystrophic muscles. As such, pharmacological interventions that target HDACs (that is, HDAC inhibitors) are of potential therapeutic interest for the treatment of muscular dystrophies. In this study, we explored the effectiveness of long-term treatment with different doses of the HDAC inhibitor givinostat in mdx mice—the mouse model of Duchenne muscular dystrophy (DMD). This study identified an efficacy for recovering functional and histological parameters within a window between 5 and 10 mg/kg/d of givinostat, with evident reduction of the beneficial effects with 1 mg/kg/d dosage. The long-term (3.5 months) exposure of 1.5-month-old mdx mice to optimal concentrations of givinostat promoted the formation of muscles with increased cross-sectional area and reduced fibrotic scars and fatty infiltration, leading to an overall improvement of endurance performance in treadmill tests and increased membrane stability. Interestingly, a reduced inflammatory infiltrate was observed in muscles of mdx mice exposed to 5 and 10 mg/kg/d of givinostat. A parallel pharmacokinetic/pharmacodynamic analysis confirmed the relationship between the effective doses of givinostat and the drug distribution in muscles and blood of treated mice. These findings provide the preclinical basis for an immediate translation of givinostat into clinical studies with DMD patients.

Notes

Acknowledgments

PL Puri is an Associate Investigator of Sanford Children’s Health Research Center. This work has been supported by the following grants to PL Puri: R01AR052779 and P30 AR061303 from the National Institute of Health/National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS), MDA, AFM, FILAS and EPIGEN. This work has benefited from research funding from the European Community’s Seventh Framework Programme in the project FP7-Health 2009 ENDOSTEM 241440 (Activation of vasculature associated stem cells and muscle stem cells for the repair and maintenance of muscle tissue). C Mozzetta was supported by an AFM fellowship. We thank A Sandri (Plaisant) for the excellent assistant in treating and monitoring in vivo functions of mdx mice.

References

  1. 1.
    Dalkilic I, Kunkel LM. (2003) Muscular dystrophies: genes to pathogenesis. Curr Opin. Genet. Dev. 13:231–8.CrossRefPubMedGoogle Scholar
  2. 2.
    Hoffman EP, Brown RH Jr, Kunkel LM. (1987) Dystrophin: the protein product of the Duchenne muscular dystrophy locus. Cell. 51:919–28.CrossRefPubMedGoogle Scholar
  3. 3.
    Mendell JR, Boué DR, Martin PT. (2006) The congenital muscular dystrophies: recent advances and molecular insights. Pediatr. Dev. Pathol. 9:427–43.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Muntoni F, Fisher I, Morgan JE, Abraham D. (2002) Steroids in Duchenne muscular dystrophy: from clinical trials to genomic research. Neuromuscul. Disord. 12(Suppl 1):S162–5.CrossRefPubMedGoogle Scholar
  5. 5.
    Consalvi S, et al. (2011) Histone deacetylase inhibitors in the treatment of muscular dystrophies: epigenetic drugs for genetic diseases. Mol. Med. 17:457–65.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Minetti GC, et al. (2006) Functional and morphological recovery of dystrophic muscles in mice treated with deacetylase inhibitors. Nat. Med. 12:1147–50.CrossRefGoogle Scholar
  7. 7.
    Colussi C, et al. (2008) HDAC2 blockade by nitric oxide and histone deacetylase inhibitors reveals a common target in Duchenne muscular dystrophy treatment. Proc. Natl. Acad. Sci. U. S. A. 105:19183–7.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Iezzi S, et al. (2004) Deacetylase inhibitors increase muscle cell size by promoting myoblast recruitment and fusion through induction of follistatin. Dev. Cell. 6:673–84.CrossRefPubMedGoogle Scholar
  9. 9.
    Dinarello CA, Fossati G, Mascagni P. (2011) Histone deacetylase inhibitors for treating a spectrum of diseases not related to cancer. Mol. Med. 17:333–52.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Colussi C, et al. (2010) Proteomic profile of differentially expressed plasma proteins from dystrophic mice and following suberoylanilide hydroxamic acid treatment. Proteomics Clin. Appl. 4:71–83.CrossRefGoogle Scholar
  11. 11.
    Vojinovic J, et al. (2011) Safety and efficacy of an oral histone deacetylase inhibitor in systemic-onset juvenile idiopathic arthritis. Arthritis Rheum. 63:1452–8.CrossRefGoogle Scholar
  12. 12.
    Vojinovic J, Damjanov N. (2011) HDAC inhibition in rheumatoid arthritis and juvenile idiopathic arthritis. Mol. Med. 17:397–403.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Committee for the Update of the Guide for the Care and Use of Laboratory Animals, Institute for Laboratory Animal Research, Division on Earth and Life Studies, National Research Council of the National Academies. (2011) Guide for the Care and Use of Laboratory Animals. 8th edition. Washington (DC): National Academies Press. [cited 2013 Apr 23]. Available from: https://doi.org/oacu.od.nih.gov/regs/Google Scholar
  14. 14.
    Iezzi S, Cossu G, Nervi C, Sartorelli V, Puri PL. (2002) Stage-specific modulation of skeletal myo-genesis by inhibitors of nuclear deacetylases. Proc. Natl. Acad. Sci. U. S. A. 99:7757–62.CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Furlan A, et al. (2011) Pharmacokinetics, safety and inducible cytokine responses during a phase 1 trial of the oral histone deacetylase inhibitor ITF2357 (givinostat). Mol. Med. 17:353–62.CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Grounds MD, et al. (2008) Towards developing standard operating procedures for pre-clinical testing in the mdx mouse model of Duchenne muscular dystrophy. Neurobiol. Dis. 31:1–19.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Vidal B, et al. (2008) Fibrinogen drives dystrophic muscle fibrosis via a TGFbeta/alternative macrophage activation pathway. Genes Dev. 22:1747–52.CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Barone FC, et al. (1991) Polymorphonuclear leukocyte infiltration into cerebral focal ischemic tissue: myeloperoxidase activity assay and histologic verification. J. Neurosci. Res. 29:336–45.CrossRefPubMedGoogle Scholar
  19. 19.
    Zhang ZG, Chopp M. (1997) Measurement of myeloperoxidase immunoreactive cells in ische-mic brain after transient middle cerebral artery occlusion in the rat. Neurosci. Res. Commun. 20:85–91.CrossRefGoogle Scholar
  20. 20.
    Leoni F, et al. (2005) The histone deacetylase inhibitor ITF2357 reduces production of pro-inflammatory cytokines in vitro and systemic inflammation in vivo. Mol. Med. 11:1–15.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Hamer PW, McGeachie JM, Davies MJ, Grounds MD (2002). Evans Blue dye as an in vivo marker of myofibre damage: optimising parameters for detecting initial myofibre membrane permeability. J. Anat. 200:69–79.CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Mozzetta C, Minetti G, Puri PL. (2009) Regenerative pharmacology in the treatment of genetic diseases: the paradigm of muscular dystrophy. Int. J. Biochem. Cell Biol. 41:701–10.CrossRefGoogle Scholar
  23. 23.
    Evans NP, Misyak SA, Robertson JL, Bassaganya-Riera J, Grange RW. (2009) Dysregulated intracellular signaling and inflammatory gene expression during initial disease onset in Duchenne muscular dystrophy. Am. J. Phys. Med. Rehabil. 88:502–22.CrossRefPubMedGoogle Scholar
  24. 24.
    Brunelli S, et al. (2007) Nitric oxide release combined with nonsteroidal antiinflammatory activity prevents muscular dystrophy pathology and enhances stem cell therapy. Proc. Natl. Acad. Sci. U. S. A. 104:264–9.CrossRefGoogle Scholar

Copyright information

© The Author(s) 2013

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, and provide a link to the Creative Commons license. You do not have permission under this license to share adapted material derived from this article or parts of it.

The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.

To view a copy of this license, visit (https://doi.org/creativecommons.org/licenses/by-nc-nd/4.0/)

Authors and Affiliations

  • Silvia Consalvi
    • 1
  • Chiara Mozzetta
    • 1
  • Paolo Bettica
    • 2
  • Massimiliano Germani
    • 3
  • Francesco Fiorentini
    • 3
  • Francesca Del Bene
    • 3
  • Maurizio Rocchetti
  • Flavio Leoni
    • 2
  • Valmen Monzani
    • 2
  • Paolo Mascagni
    • 2
  • Pier Lorenzo Puri
    • 1
    • 4
  • Valentina Saccone
    • 1
  1. 1.IRCCS Fondazione Santa LuciaRomeItaly
  2. 2.Italfarmaco SpAMilanItaly
  3. 3.Accelera SpAMilanItaly
  4. 4.Sanford-Burnham Medical Research InstituteSanford Children’s Health Research CenterLa JollaUSA

Personalised recommendations