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Spiruchostatin A inhibits proliferation and differentiation of fibroblasts from patients with pulmonary fibrosis

Spiruchostatin A inhibits proliferation and differentiation of fibroblasts from patients with pulmonary fibrosis
Spiruchostatin A inhibits proliferation and differentiation of fibroblasts from patients with pulmonary fibrosis
Idiopathic pulmonary fibrosis (IPF) is a progressive scarring disorder characterized by the proliferation of interstitial fibroblasts and the deposition of extracellular matrix causing impaired gas exchange. Spiruchostatin A (SpA) is a histone deacetylase inhibitor (HDI) with selectivity toward Class I enzymes, which distinguishes it from other nonspecific HDIs that are reported to inhibit (myo)fibroblast proliferation and differentiation. Because the selectivity of HDIs may be important clinically, we postulated that SpA inhibits the proliferation and differentiation of IPF fibroblasts. Primary fibroblasts were grown from lung biopsy explants obtained from patients with IPF or from normal control subjects, using two-dimensional or three-dimensional culture models. The effect of SpA on fibroproliferation in serum-containing medium ± transforming growth factor (TGF)–?1 was quantified by methylene blue binding. The acetylation of histone H3, the expression of the cell-cycle inhibitor p21waf1, and the myofibroblast markers ?–smooth muscle actin (?-SMA) and collagens I and III were determined by Western blotting, quantitative RT-PCR, immunofluorescent staining, or colorimetry. SpA inhibited the proliferation of IPF or normal fibroblasts in a time-dependent and concentration-dependent manner (concentration required to achieve 50% inhibition = 3.8 ± 0.4 nM versus 7.8 ± 0.2 nM, respectively; P < 0.05), with little cytotoxicity. Western blot analyses revealed that SpA caused a concentration-dependent increase in histone H3 acetylation, paralleling its antiproliferative effect. SpA also increased p21waf1 expression, suggesting that direct cell-cycle regulation was the mechanism of inhibiting proliferation. Although treatment with TGF-?1 induced myofibroblast differentiation associated with increased expression of ?-SMA, collagen I and collagen III and soluble collagen release, these responses were potently inhibited by SpA. These data support the concept that bicyclic tetrapeptide HDIs merit further investigation as potential treatments for IPF.
1044-1549
687-694
Davies, Elizabeth R.
aff6b3f7-91e0-4fd3-9554-a9389024b63b
Haitchi, Hans Michael
6efc50e5-4dbe-414c-8f8b-b261ce1fc5c4
Thatcher, Thomas H.
b4138c92-d1b3-41a8-9991-3374f68ac516
Sime, Patricia J.
d96733e9-a51c-4dc0-90c0-203e550827d1
Kottmann, R. Matthew
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Ganesan, Arasu
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Packham, Graham
fdabe56f-2c58-469c-aadf-38878f233394
O'Reilly, Katherine M.A.
6c323cd0-0c68-4913-9666-7e4f655b816f
Davies, Donna E.
7de8fdc7-3640-4e3a-aa91-d0e03f990c38
Davies, Elizabeth R.
aff6b3f7-91e0-4fd3-9554-a9389024b63b
Haitchi, Hans Michael
6efc50e5-4dbe-414c-8f8b-b261ce1fc5c4
Thatcher, Thomas H.
b4138c92-d1b3-41a8-9991-3374f68ac516
Sime, Patricia J.
d96733e9-a51c-4dc0-90c0-203e550827d1
Kottmann, R. Matthew
f6c7ebf6-80d2-489a-b150-98fe9cf5e2ea
Ganesan, Arasu
b089a6eb-86e6-4f65-878d-4766dfdeb6b7
Packham, Graham
fdabe56f-2c58-469c-aadf-38878f233394
O'Reilly, Katherine M.A.
6c323cd0-0c68-4913-9666-7e4f655b816f
Davies, Donna E.
7de8fdc7-3640-4e3a-aa91-d0e03f990c38

Davies, Elizabeth R., Haitchi, Hans Michael, Thatcher, Thomas H., Sime, Patricia J., Kottmann, R. Matthew, Ganesan, Arasu, Packham, Graham, O'Reilly, Katherine M.A. and Davies, Donna E. (2012) Spiruchostatin A inhibits proliferation and differentiation of fibroblasts from patients with pulmonary fibrosis. American Journal of Respiratory Cell and Molecular Biology, 46 (5), 687-694. (doi:10.1165/rcmb.2011-0040OC). (PMID:22246864)

Record type: Article

Abstract

Idiopathic pulmonary fibrosis (IPF) is a progressive scarring disorder characterized by the proliferation of interstitial fibroblasts and the deposition of extracellular matrix causing impaired gas exchange. Spiruchostatin A (SpA) is a histone deacetylase inhibitor (HDI) with selectivity toward Class I enzymes, which distinguishes it from other nonspecific HDIs that are reported to inhibit (myo)fibroblast proliferation and differentiation. Because the selectivity of HDIs may be important clinically, we postulated that SpA inhibits the proliferation and differentiation of IPF fibroblasts. Primary fibroblasts were grown from lung biopsy explants obtained from patients with IPF or from normal control subjects, using two-dimensional or three-dimensional culture models. The effect of SpA on fibroproliferation in serum-containing medium ± transforming growth factor (TGF)–?1 was quantified by methylene blue binding. The acetylation of histone H3, the expression of the cell-cycle inhibitor p21waf1, and the myofibroblast markers ?–smooth muscle actin (?-SMA) and collagens I and III were determined by Western blotting, quantitative RT-PCR, immunofluorescent staining, or colorimetry. SpA inhibited the proliferation of IPF or normal fibroblasts in a time-dependent and concentration-dependent manner (concentration required to achieve 50% inhibition = 3.8 ± 0.4 nM versus 7.8 ± 0.2 nM, respectively; P < 0.05), with little cytotoxicity. Western blot analyses revealed that SpA caused a concentration-dependent increase in histone H3 acetylation, paralleling its antiproliferative effect. SpA also increased p21waf1 expression, suggesting that direct cell-cycle regulation was the mechanism of inhibiting proliferation. Although treatment with TGF-?1 induced myofibroblast differentiation associated with increased expression of ?-SMA, collagen I and collagen III and soluble collagen release, these responses were potently inhibited by SpA. These data support the concept that bicyclic tetrapeptide HDIs merit further investigation as potential treatments for IPF.

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More information

e-pub ahead of print date: January 2012
Published date: May 2012
Organisations: Clinical & Experimental Sciences

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Local EPrints ID: 341591
URI: http://eprints.soton.ac.uk/id/eprint/341591
ISSN: 1044-1549
PURE UUID: 11188c8e-bc9b-4a37-b7ae-e0bbf790a3e4
ORCID for Elizabeth R. Davies: ORCID iD orcid.org/0000-0002-8629-8324
ORCID for Graham Packham: ORCID iD orcid.org/0000-0002-9232-5691
ORCID for Donna E. Davies: ORCID iD orcid.org/0000-0002-5117-2991

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Date deposited: 30 Jul 2012 10:29
Last modified: 23 Jul 2022 02:01

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Contributors

Author: Hans Michael Haitchi
Author: Thomas H. Thatcher
Author: Patricia J. Sime
Author: R. Matthew Kottmann
Author: Arasu Ganesan
Author: Graham Packham ORCID iD
Author: Katherine M.A. O'Reilly
Author: Donna E. Davies ORCID iD

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