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Regulation of a disintegrin and metalloprotease-33 expression by transforming growth factor-?

Regulation of a disintegrin and metalloprotease-33 expression by transforming growth factor-?
Regulation of a disintegrin and metalloprotease-33 expression by transforming growth factor-?
The asthma susceptibility gene, ADAM33, is selectively expressed in mesenchymal cells and the activity of soluble ADAM33 has been linked to angiogenesis and airway remodeling. TGF-? is a profibrogenic growth factor whose expression is increased in asthma and recent studies show that it enhances shedding of soluble ADAM33. In this study, we hypothesized that TGF-? also affects ADAM33 expression in bronchial fibroblasts in asthma. Primary fibroblasts were grown from bronchial biopsies from normal and asthmatic donors and treated with TGF-?(2) to induce myofibroblast differentiation. ADAM33 expression was assessed using quantitative RT-PCR and Western blotting. To examine the mechanisms whereby TGF-?(2) affected ADAM33 expression, quantitative methylation-sensitive PCR, chromatin immunoprecipitation and nuclear accessibility assays were conducted on the ADAM33 promoter. We found that TGF-?(2) caused a time- and concentration-dependent reduction in ADAM33 mRNA expression in normal and asthmatic fibroblasts, affecting levels of splice variants similarly. TGF-?(2) also induced ADAM33 protein turnover and appearance of a cell associated C-terminal fragment. TGF-?(2) down-regulated ADAM33 mRNA expression by causing chromatin condensation around the ADAM33 promoter with deacetylation of histone H3, demethylation of H3 on lysine-4, and hypermethylation of H3 on lysine-9. However, the methylation status of the ADAM33 promoter did not change. Together these data suggest that TGF-?(2) suppresses expression of ADAM33 mRNA in normal or asthmatic fibroblasts. This occurs by altering chromatin structure, rather than by gene silencing through DNA methylation as in epithelial cells. This may provide a mechanism to finely regulate levels of ADAM33 expression in fibroblasts and may self-limit TGF-?(2)-induced ectodomain shedding of ADAM33.
a disintegrin and metalloprotease-33, myofibroblast, transforming growth factor-?, histone modification
1044-1549
633-640
Yang, Youwen
88415c3b-4613-4730-be20-8423ccae345a
Wicks, James
6b178ed1-5c0c-448c-97f1-5c505d1e530a
Haitchi, Hans Michael
68dadb29-305d-4236-884f-e9c93f4d78fe
Powell, Robert M.
884d6594-3f50-4be5-9516-d64b29dad63d
Manuyakorn, Wiparat
8b59f414-dce0-4af5-93fd-93884e36a416
Howarth, Peter H.
ff19c8c4-86b0-4a88-8f76-b3d87f142a21
Holgate, Stephen T.
2e7c17a9-6796-436e-8772-1fe6d2ac5edc
Davies, Donna E.
7de8fdc7-3640-4e3a-aa91-d0e03f990c38
Yang, Youwen
88415c3b-4613-4730-be20-8423ccae345a
Wicks, James
6b178ed1-5c0c-448c-97f1-5c505d1e530a
Haitchi, Hans Michael
68dadb29-305d-4236-884f-e9c93f4d78fe
Powell, Robert M.
884d6594-3f50-4be5-9516-d64b29dad63d
Manuyakorn, Wiparat
8b59f414-dce0-4af5-93fd-93884e36a416
Howarth, Peter H.
ff19c8c4-86b0-4a88-8f76-b3d87f142a21
Holgate, Stephen T.
2e7c17a9-6796-436e-8772-1fe6d2ac5edc
Davies, Donna E.
7de8fdc7-3640-4e3a-aa91-d0e03f990c38

Yang, Youwen, Wicks, James, Haitchi, Hans Michael, Powell, Robert M., Manuyakorn, Wiparat, Howarth, Peter H., Holgate, Stephen T. and Davies, Donna E. (2012) Regulation of a disintegrin and metalloprotease-33 expression by transforming growth factor-? American Journal of Respiratory Cell and Molecular Biology, 46 (5), 633-640. (doi:10.1165/rcmb.2011-0030OC). (PMID:22227561)

Record type: Article

Abstract

The asthma susceptibility gene, ADAM33, is selectively expressed in mesenchymal cells and the activity of soluble ADAM33 has been linked to angiogenesis and airway remodeling. TGF-? is a profibrogenic growth factor whose expression is increased in asthma and recent studies show that it enhances shedding of soluble ADAM33. In this study, we hypothesized that TGF-? also affects ADAM33 expression in bronchial fibroblasts in asthma. Primary fibroblasts were grown from bronchial biopsies from normal and asthmatic donors and treated with TGF-?(2) to induce myofibroblast differentiation. ADAM33 expression was assessed using quantitative RT-PCR and Western blotting. To examine the mechanisms whereby TGF-?(2) affected ADAM33 expression, quantitative methylation-sensitive PCR, chromatin immunoprecipitation and nuclear accessibility assays were conducted on the ADAM33 promoter. We found that TGF-?(2) caused a time- and concentration-dependent reduction in ADAM33 mRNA expression in normal and asthmatic fibroblasts, affecting levels of splice variants similarly. TGF-?(2) also induced ADAM33 protein turnover and appearance of a cell associated C-terminal fragment. TGF-?(2) down-regulated ADAM33 mRNA expression by causing chromatin condensation around the ADAM33 promoter with deacetylation of histone H3, demethylation of H3 on lysine-4, and hypermethylation of H3 on lysine-9. However, the methylation status of the ADAM33 promoter did not change. Together these data suggest that TGF-?(2) suppresses expression of ADAM33 mRNA in normal or asthmatic fibroblasts. This occurs by altering chromatin structure, rather than by gene silencing through DNA methylation as in epithelial cells. This may provide a mechanism to finely regulate levels of ADAM33 expression in fibroblasts and may self-limit TGF-?(2)-induced ectodomain shedding of ADAM33.

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More information

e-pub ahead of print date: 6 January 2012
Published date: May 2012
Keywords: a disintegrin and metalloprotease-33, myofibroblast, transforming growth factor-?, histone modification
Organisations: Clinical & Experimental Sciences

Identifiers

Local EPrints ID: 337451
URI: http://eprints.soton.ac.uk/id/eprint/337451
ISSN: 1044-1549
PURE UUID: ce813c39-eedc-479c-89b7-e28efc2b6786
ORCID for Hans Michael Haitchi: ORCID iD orcid.org/0000-0001-8603-302X
ORCID for Donna E. Davies: ORCID iD orcid.org/0000-0002-5117-2991

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Date deposited: 26 Apr 2012 08:56
Last modified: 15 Mar 2024 03:14

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Contributors

Author: Youwen Yang
Author: James Wicks
Author: Robert M. Powell
Author: Wiparat Manuyakorn
Author: Donna E. Davies ORCID iD

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