Tissue factor-thrombin signaling enhances the fibrotic activity of myofibroblasts in systemic sclerosis through up-regulation of endothelin receptor A

Akrivi Chrysanthopoulou; Ioannis Mitroulis; Konstantinos Kambas; Panagiotis Skendros; Ioannis Kourtzelis; Stergios Vradelis; George Kolios; Spyros Aslanidis; Michael Doumas; Konstantinos Ritis

doi:10.1002/art.30586

Tissue factor-thrombin signaling enhances the fibrotic activity of myofibroblasts in systemic sclerosis through up-regulation of endothelin receptor A

Akrivi Chrysanthopoulou, Ioannis Mitroulis, Konstantinos Kambas, Panagiotis Skendros, Ioannis Kourtzelis, Stergios Vradelis, George Kolios, Spyros Aslanidis, Michael Doumas, Konstantinos Ritis

The Hull York Medical School

Research output: Contribution to journal › Article › peer-review

Abstract

OBJECTIVE: The extrinsic coagulation cascade is involved in the fibrotic process, via thrombin-dependent induction of CCN2 (connective tissue growth factor) expression. Given the previously reported activation of the coagulation system in systemic sclerosis (SSc), we undertook the present study to investigate the involvement of cross-talk between the tissue factor (TF)-thrombin axis and endothelin 1 (ET-1) signaling in the fibrotic activity of SSc.

METHODS: Human colonic myofibroblasts (HCMFs) from 6 patients with SSc and gastrointestinal symptoms and from 6 control subjects were isolated and cultured under various conditions. Messenger RNA and protein levels of TF, CCN2, and endothelin receptor A (ET(A) ) were investigated. Collagen production and migratory activity of HCMFs were further assessed.

RESULTS: HCMFs from SSc patients demonstrated increased basal CCN2 production, collagen deposition, and migration rate, in a thrombin-dependent manner. Increased TF expression was also observed in SSc HCMFs. Subsequent activation of the extrinsic coagulation system resulted in thrombin-dependent enhancement of ET(A) expression. ET(A) overexpression led to further increases in both TF expression and fibrotic activity in HCMFs. Moreover, inhibition of ET-1 signaling by bosentan abolished the TF-mediated fibrotic capacity of HCMFs.

CONCLUSION: Tissue factor-thrombin signaling is involved in the increased fibrotic activity of HCMFs from patients with SSc. Moreover, the up-regulation of ET(A) expression by thrombin and the effect of ET-1 in the induction of TF expression indicate an amplification loop for enhanced collagen deposition. Therapeutic interventions targeting the extrinsic coagulation system or ET-1 signaling may provide clinical benefit by breaking this vicious circle.

Original language	English
Pages (from-to)	3586-97
Number of pages	12
Journal	Arthritis and rheumatism
Volume	63
Issue number	11
DOIs	https://doi.org/10.1002/art.30586
Publication status	Published - Nov 2011

Bibliographical note

Keywords

Cell Movement
Cell Proliferation
Collagen Type I/metabolism
Colon/metabolism
Connective Tissue Growth Factor/metabolism
Fibrosis
Humans
Myofibroblasts/metabolism
Receptor, Endothelin A/metabolism
Receptor, PAR-1/metabolism
Scleroderma, Systemic/metabolism
Signal Transduction
Thrombin/metabolism
Thromboplastin/metabolism
Up-Regulation

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Cite this

Chrysanthopoulou, A., Mitroulis, I., Kambas, K., Skendros, P., Kourtzelis, I., Vradelis, S., Kolios, G., Aslanidis, S., Doumas, M., & Ritis, K. (2011). Tissue factor-thrombin signaling enhances the fibrotic activity of myofibroblasts in systemic sclerosis through up-regulation of endothelin receptor A. Arthritis and rheumatism, 63(11), 3586-97. https://doi.org/10.1002/art.30586

@article{cc9edabb150e4f4fa092962e287fa11e,

title = "Tissue factor-thrombin signaling enhances the fibrotic activity of myofibroblasts in systemic sclerosis through up-regulation of endothelin receptor A",

abstract = "OBJECTIVE: The extrinsic coagulation cascade is involved in the fibrotic process, via thrombin-dependent induction of CCN2 (connective tissue growth factor) expression. Given the previously reported activation of the coagulation system in systemic sclerosis (SSc), we undertook the present study to investigate the involvement of cross-talk between the tissue factor (TF)-thrombin axis and endothelin 1 (ET-1) signaling in the fibrotic activity of SSc.METHODS: Human colonic myofibroblasts (HCMFs) from 6 patients with SSc and gastrointestinal symptoms and from 6 control subjects were isolated and cultured under various conditions. Messenger RNA and protein levels of TF, CCN2, and endothelin receptor A (ET(A) ) were investigated. Collagen production and migratory activity of HCMFs were further assessed.RESULTS: HCMFs from SSc patients demonstrated increased basal CCN2 production, collagen deposition, and migration rate, in a thrombin-dependent manner. Increased TF expression was also observed in SSc HCMFs. Subsequent activation of the extrinsic coagulation system resulted in thrombin-dependent enhancement of ET(A) expression. ET(A) overexpression led to further increases in both TF expression and fibrotic activity in HCMFs. Moreover, inhibition of ET-1 signaling by bosentan abolished the TF-mediated fibrotic capacity of HCMFs.CONCLUSION: Tissue factor-thrombin signaling is involved in the increased fibrotic activity of HCMFs from patients with SSc. Moreover, the up-regulation of ET(A) expression by thrombin and the effect of ET-1 in the induction of TF expression indicate an amplification loop for enhanced collagen deposition. Therapeutic interventions targeting the extrinsic coagulation system or ET-1 signaling may provide clinical benefit by breaking this vicious circle.",

keywords = "Cell Movement, Cell Proliferation, Collagen Type I/metabolism, Colon/metabolism, Connective Tissue Growth Factor/metabolism, Fibrosis, Humans, Myofibroblasts/metabolism, Receptor, Endothelin A/metabolism, Receptor, PAR-1/metabolism, Scleroderma, Systemic/metabolism, Signal Transduction, Thrombin/metabolism, Thromboplastin/metabolism, Up-Regulation",

author = "Akrivi Chrysanthopoulou and Ioannis Mitroulis and Konstantinos Kambas and Panagiotis Skendros and Ioannis Kourtzelis and Stergios Vradelis and George Kolios and Spyros Aslanidis and Michael Doumas and Konstantinos Ritis",

note = "Copyright {\textcopyright} 2011 by the American College of Rheumatology.",

year = "2011",

month = nov,

doi = "10.1002/art.30586",

language = "English",

volume = "63",

pages = "3586--97",

journal = "Arthritis and rheumatism",

issn = "0004-3591",

publisher = "Wiley-Blackwell",

number = "11",

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Chrysanthopoulou, A, Mitroulis, I, Kambas, K, Skendros, P, Kourtzelis, I, Vradelis, S, Kolios, G, Aslanidis, S, Doumas, M & Ritis, K 2011, 'Tissue factor-thrombin signaling enhances the fibrotic activity of myofibroblasts in systemic sclerosis through up-regulation of endothelin receptor A', Arthritis and rheumatism, vol. 63, no. 11, pp. 3586-97. https://doi.org/10.1002/art.30586

TY - JOUR

T1 - Tissue factor-thrombin signaling enhances the fibrotic activity of myofibroblasts in systemic sclerosis through up-regulation of endothelin receptor A

AU - Chrysanthopoulou, Akrivi

AU - Mitroulis, Ioannis

AU - Kambas, Konstantinos

AU - Skendros, Panagiotis

AU - Kourtzelis, Ioannis

AU - Vradelis, Stergios

AU - Kolios, George

AU - Aslanidis, Spyros

AU - Doumas, Michael

AU - Ritis, Konstantinos

PY - 2011/11

Y1 - 2011/11

N2 - OBJECTIVE: The extrinsic coagulation cascade is involved in the fibrotic process, via thrombin-dependent induction of CCN2 (connective tissue growth factor) expression. Given the previously reported activation of the coagulation system in systemic sclerosis (SSc), we undertook the present study to investigate the involvement of cross-talk between the tissue factor (TF)-thrombin axis and endothelin 1 (ET-1) signaling in the fibrotic activity of SSc.METHODS: Human colonic myofibroblasts (HCMFs) from 6 patients with SSc and gastrointestinal symptoms and from 6 control subjects were isolated and cultured under various conditions. Messenger RNA and protein levels of TF, CCN2, and endothelin receptor A (ET(A) ) were investigated. Collagen production and migratory activity of HCMFs were further assessed.RESULTS: HCMFs from SSc patients demonstrated increased basal CCN2 production, collagen deposition, and migration rate, in a thrombin-dependent manner. Increased TF expression was also observed in SSc HCMFs. Subsequent activation of the extrinsic coagulation system resulted in thrombin-dependent enhancement of ET(A) expression. ET(A) overexpression led to further increases in both TF expression and fibrotic activity in HCMFs. Moreover, inhibition of ET-1 signaling by bosentan abolished the TF-mediated fibrotic capacity of HCMFs.CONCLUSION: Tissue factor-thrombin signaling is involved in the increased fibrotic activity of HCMFs from patients with SSc. Moreover, the up-regulation of ET(A) expression by thrombin and the effect of ET-1 in the induction of TF expression indicate an amplification loop for enhanced collagen deposition. Therapeutic interventions targeting the extrinsic coagulation system or ET-1 signaling may provide clinical benefit by breaking this vicious circle.

AB - OBJECTIVE: The extrinsic coagulation cascade is involved in the fibrotic process, via thrombin-dependent induction of CCN2 (connective tissue growth factor) expression. Given the previously reported activation of the coagulation system in systemic sclerosis (SSc), we undertook the present study to investigate the involvement of cross-talk between the tissue factor (TF)-thrombin axis and endothelin 1 (ET-1) signaling in the fibrotic activity of SSc.METHODS: Human colonic myofibroblasts (HCMFs) from 6 patients with SSc and gastrointestinal symptoms and from 6 control subjects were isolated and cultured under various conditions. Messenger RNA and protein levels of TF, CCN2, and endothelin receptor A (ET(A) ) were investigated. Collagen production and migratory activity of HCMFs were further assessed.RESULTS: HCMFs from SSc patients demonstrated increased basal CCN2 production, collagen deposition, and migration rate, in a thrombin-dependent manner. Increased TF expression was also observed in SSc HCMFs. Subsequent activation of the extrinsic coagulation system resulted in thrombin-dependent enhancement of ET(A) expression. ET(A) overexpression led to further increases in both TF expression and fibrotic activity in HCMFs. Moreover, inhibition of ET-1 signaling by bosentan abolished the TF-mediated fibrotic capacity of HCMFs.CONCLUSION: Tissue factor-thrombin signaling is involved in the increased fibrotic activity of HCMFs from patients with SSc. Moreover, the up-regulation of ET(A) expression by thrombin and the effect of ET-1 in the induction of TF expression indicate an amplification loop for enhanced collagen deposition. Therapeutic interventions targeting the extrinsic coagulation system or ET-1 signaling may provide clinical benefit by breaking this vicious circle.

KW - Cell Movement

KW - Cell Proliferation

KW - Collagen Type I/metabolism

KW - Colon/metabolism

KW - Connective Tissue Growth Factor/metabolism

KW - Fibrosis

KW - Humans

KW - Myofibroblasts/metabolism

KW - Receptor, Endothelin A/metabolism

KW - Receptor, PAR-1/metabolism

KW - Scleroderma, Systemic/metabolism

KW - Signal Transduction

KW - Thrombin/metabolism

KW - Thromboplastin/metabolism

KW - Up-Regulation

U2 - 10.1002/art.30586

DO - 10.1002/art.30586

M3 - Article

C2 - 21834070

SN - 0004-3591

VL - 63

SP - 3586

EP - 3597

JO - Arthritis and rheumatism

JF - Arthritis and rheumatism

IS - 11

ER -