The Impact of Hydroxylase Inhibition on TGF-β1 Induced Fibrosis Associated with Inflammatory Bowel Disease

Mario C Manresa, Murtaza Tambuwala, Miguel Cavadas, Alex Cheong, Eoin Cummins, Cormac Taylor

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

Fibrosis is a complication of chronic inflammatory disorders such as inflammatory bowel disease (IBD). Fibrosis is responsible for 75% of surgical interventions in IBD. Pharmacological inhibition of O2 sensing hydroxylases ameliorates inflammation in models of IBD. We hypothesized that hydroxylase inhibition may also impact fibrosis in IBD. To test this, the effect of the hydroxylase inhibitor DMOG was investigated. Mice exposed to 2.5% dextran sodium sulphate in drinking water for 5 days and allowed to recover for 14 days, developed colon fibrosis reflected by an altered pattern of submucosal collagen deposition, increased fibroblast infiltration and higher expression of Smad3. Mice treated with DMOG exhibited lower submucosal collagen accumulation, reduced fibroblast infiltration and less Smad3 positive staining. An in vitro model was then used to investigate the mechanism of the anti-fibrotic action of DMOG. Fibroblasts were treated with TGF-β1, key mediator of fibrosis. DMOG was given to the cells prior to TGF-β1, and α-smooth muscle actin (α-SMA) and collagen-I were analyzed as markers of fibrosis. TGF-β1 induced phenotypic changes associated to higher expression of collagen-I and α-SMA. Both markers were reduced in DMOG treated cells, suggesting an inhibition of TGF-β1 mediated fibroblast activation. We further investigated the impact of DMOG upon TGF-β1 signalling. DMOG did not affect Smad2/3 (canonical) TGF dependent phosphorylation. In contrast, it reduced TGF-β1 dependent ERK (non canonical) phosphorylation. In conclusion, hydroxylase inhibition reduces fibrosis in vitro and in vivo, at least in part, through the inhibition of non canonical TGF-β signalling.
LanguageEnglish
Title of host publicationUnknown Host Publication
Number of pages1
Volume29
Publication statusPublished - 10 Apr 2015
EventHypoxia: From Basic Mechanisms to Therapeutics -
Duration: 10 Apr 2015 → …

Conference

ConferenceHypoxia: From Basic Mechanisms to Therapeutics
Period10/04/15 → …

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Mixed Function Oxygenases
Inflammatory Bowel Diseases
Fibrosis
Collagen
Fibroblasts
Smooth Muscle
Actins
Phosphorylation
Dextran Sulfate
Transforming Growth Factor alpha
Drinking Water
Transforming Growth Factor beta
Colon
Pharmacology
Staining and Labeling
Inflammation

Keywords

  • Fibrosis
  • Inflammatory bowel disease
  • Hydroxylases

Cite this

Manresa, M. C., Tambuwala, M., Cavadas, M., Cheong, A., Cummins, E., & Taylor, C. (2015). The Impact of Hydroxylase Inhibition on TGF-β1 Induced Fibrosis Associated with Inflammatory Bowel Disease. In Unknown Host Publication (Vol. 29)
Manresa, Mario C ; Tambuwala, Murtaza ; Cavadas, Miguel ; Cheong, Alex ; Cummins, Eoin ; Taylor, Cormac. / The Impact of Hydroxylase Inhibition on TGF-β1 Induced Fibrosis Associated with Inflammatory Bowel Disease. Unknown Host Publication. Vol. 29 2015.
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abstract = "Fibrosis is a complication of chronic inflammatory disorders such as inflammatory bowel disease (IBD). Fibrosis is responsible for 75{\%} of surgical interventions in IBD. Pharmacological inhibition of O2 sensing hydroxylases ameliorates inflammation in models of IBD. We hypothesized that hydroxylase inhibition may also impact fibrosis in IBD. To test this, the effect of the hydroxylase inhibitor DMOG was investigated. Mice exposed to 2.5{\%} dextran sodium sulphate in drinking water for 5 days and allowed to recover for 14 days, developed colon fibrosis reflected by an altered pattern of submucosal collagen deposition, increased fibroblast infiltration and higher expression of Smad3. Mice treated with DMOG exhibited lower submucosal collagen accumulation, reduced fibroblast infiltration and less Smad3 positive staining. An in vitro model was then used to investigate the mechanism of the anti-fibrotic action of DMOG. Fibroblasts were treated with TGF-β1, key mediator of fibrosis. DMOG was given to the cells prior to TGF-β1, and α-smooth muscle actin (α-SMA) and collagen-I were analyzed as markers of fibrosis. TGF-β1 induced phenotypic changes associated to higher expression of collagen-I and α-SMA. Both markers were reduced in DMOG treated cells, suggesting an inhibition of TGF-β1 mediated fibroblast activation. We further investigated the impact of DMOG upon TGF-β1 signalling. DMOG did not affect Smad2/3 (canonical) TGF dependent phosphorylation. In contrast, it reduced TGF-β1 dependent ERK (non canonical) phosphorylation. In conclusion, hydroxylase inhibition reduces fibrosis in vitro and in vivo, at least in part, through the inhibition of non canonical TGF-β signalling.",
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Manresa, MC, Tambuwala, M, Cavadas, M, Cheong, A, Cummins, E & Taylor, C 2015, The Impact of Hydroxylase Inhibition on TGF-β1 Induced Fibrosis Associated with Inflammatory Bowel Disease. in Unknown Host Publication. vol. 29, Hypoxia: From Basic Mechanisms to Therapeutics, 10/04/15.

The Impact of Hydroxylase Inhibition on TGF-β1 Induced Fibrosis Associated with Inflammatory Bowel Disease. / Manresa, Mario C; Tambuwala, Murtaza; Cavadas, Miguel; Cheong, Alex; Cummins, Eoin; Taylor, Cormac.

Unknown Host Publication. Vol. 29 2015.

Research output: Chapter in Book/Report/Conference proceedingConference contribution

TY - GEN

T1 - The Impact of Hydroxylase Inhibition on TGF-β1 Induced Fibrosis Associated with Inflammatory Bowel Disease

AU - Manresa, Mario C

AU - Tambuwala, Murtaza

AU - Cavadas, Miguel

AU - Cheong, Alex

AU - Cummins, Eoin

AU - Taylor, Cormac

PY - 2015/4/10

Y1 - 2015/4/10

N2 - Fibrosis is a complication of chronic inflammatory disorders such as inflammatory bowel disease (IBD). Fibrosis is responsible for 75% of surgical interventions in IBD. Pharmacological inhibition of O2 sensing hydroxylases ameliorates inflammation in models of IBD. We hypothesized that hydroxylase inhibition may also impact fibrosis in IBD. To test this, the effect of the hydroxylase inhibitor DMOG was investigated. Mice exposed to 2.5% dextran sodium sulphate in drinking water for 5 days and allowed to recover for 14 days, developed colon fibrosis reflected by an altered pattern of submucosal collagen deposition, increased fibroblast infiltration and higher expression of Smad3. Mice treated with DMOG exhibited lower submucosal collagen accumulation, reduced fibroblast infiltration and less Smad3 positive staining. An in vitro model was then used to investigate the mechanism of the anti-fibrotic action of DMOG. Fibroblasts were treated with TGF-β1, key mediator of fibrosis. DMOG was given to the cells prior to TGF-β1, and α-smooth muscle actin (α-SMA) and collagen-I were analyzed as markers of fibrosis. TGF-β1 induced phenotypic changes associated to higher expression of collagen-I and α-SMA. Both markers were reduced in DMOG treated cells, suggesting an inhibition of TGF-β1 mediated fibroblast activation. We further investigated the impact of DMOG upon TGF-β1 signalling. DMOG did not affect Smad2/3 (canonical) TGF dependent phosphorylation. In contrast, it reduced TGF-β1 dependent ERK (non canonical) phosphorylation. In conclusion, hydroxylase inhibition reduces fibrosis in vitro and in vivo, at least in part, through the inhibition of non canonical TGF-β signalling.

AB - Fibrosis is a complication of chronic inflammatory disorders such as inflammatory bowel disease (IBD). Fibrosis is responsible for 75% of surgical interventions in IBD. Pharmacological inhibition of O2 sensing hydroxylases ameliorates inflammation in models of IBD. We hypothesized that hydroxylase inhibition may also impact fibrosis in IBD. To test this, the effect of the hydroxylase inhibitor DMOG was investigated. Mice exposed to 2.5% dextran sodium sulphate in drinking water for 5 days and allowed to recover for 14 days, developed colon fibrosis reflected by an altered pattern of submucosal collagen deposition, increased fibroblast infiltration and higher expression of Smad3. Mice treated with DMOG exhibited lower submucosal collagen accumulation, reduced fibroblast infiltration and less Smad3 positive staining. An in vitro model was then used to investigate the mechanism of the anti-fibrotic action of DMOG. Fibroblasts were treated with TGF-β1, key mediator of fibrosis. DMOG was given to the cells prior to TGF-β1, and α-smooth muscle actin (α-SMA) and collagen-I were analyzed as markers of fibrosis. TGF-β1 induced phenotypic changes associated to higher expression of collagen-I and α-SMA. Both markers were reduced in DMOG treated cells, suggesting an inhibition of TGF-β1 mediated fibroblast activation. We further investigated the impact of DMOG upon TGF-β1 signalling. DMOG did not affect Smad2/3 (canonical) TGF dependent phosphorylation. In contrast, it reduced TGF-β1 dependent ERK (non canonical) phosphorylation. In conclusion, hydroxylase inhibition reduces fibrosis in vitro and in vivo, at least in part, through the inhibition of non canonical TGF-β signalling.

KW - Fibrosis

KW - Inflammatory bowel disease

KW - Hydroxylases

M3 - Conference contribution

VL - 29

BT - Unknown Host Publication

ER -

Manresa MC, Tambuwala M, Cavadas M, Cheong A, Cummins E, Taylor C. The Impact of Hydroxylase Inhibition on TGF-β1 Induced Fibrosis Associated with Inflammatory Bowel Disease. In Unknown Host Publication. Vol. 29. 2015