Involvement of CTGF in TGF-β1–Stimulation of Myofibroblast Differentiation and Collagen Matrix Contraction in the Presence of Mechanical Stress

Garrett, Qian; Khaw, PT; Blalock, Timothy D.; Schultz, Gregory S.; Grotendorst, Gary R.; Daniels, Julie T.

doi:10.1167/iovs.03-0660

Cited by 124 publications

(111 citation statements)

References 38 publications

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“…[51][52][53][54][55][56] TGF-b1 can induce fibroblast-to-myofibroblast differentiation. 12,[57][58][59] In 2008, Wells and Discher determined that the matrix stiffness was related with TGFb1-induced myofibroblast differentiation, 60 in which they showed that higher tension could increase the release of active TGF-b1, resulting in myofibroblast differentiation. In the present study, we observed that TGF-b1 induced myofibroblast differentiation in standard monolayer culture condition at both the gene and protein levels.…”

Section: D Effects Of Tgf-b1mentioning

confidence: 99%

Response of Fibroblasts to Transforming Growth Factor-β1 on Two-Dimensional and in Three-Dimensional Hyaluronan Hydrogels

Chen

Thibeault

2012

Tissue Engineering Part A

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Transforming growth factor-b1 (TGF-b1), an important cytokine with multiple functions, is secreted during wound healing. Previous studies have utilized two-dimensional (2D) cell culture to elucidate the functions of TGF-b1; however, 2D culture does not represent the complex three-dimensional (3D) in vivo environment. Using a synthetic hyaluronan (HA) extracellular matrix (ECM) hydrogel, we investigated the effect of TGF-b1 on fibroblasts cultured in three conditions-on tissue culture polystyrene (TCP), on HA (2D), and in HA (3D). After TGF-b1 treatment (0.1 to 20 ng/mL), morphological features and ECM regulation were analyzed by immunocytochemistry, Western blot, quantitative polymerase chain reaction, and zymogram assays. On TCP, cells showed the typical spindle shape with strong alpha smooth muscle actin (a-SMA) staining of cytoplasmic myofilaments along the cell axes after TGF-b1 treatment; on HA (2D), spindle-shape cells showed little a-SMA staining; in HA (3D), cells were smaller and rounded with less a-SMA deposition. The a-SMA gene and protein expression on TCP were significantly upregulated by TGF-b1, but TGF-b1 did not induce a-SMA expression in the presence of HA (both 2D and 3D). 3D HA culture significantly downregulated collagen I, III, and fibronectin expression, increased matrix metalloproteinase 1 and 2 (MMP1/MMP2) activity, upregulated MMP1 mRNA and downregulated TIMP3 mRNA expression. This study suggested that exogenous HA, particularly in 3D culture, appears to suppress ECM production, enhances ECM degradation and remodeling, and inhibits myofibroblast differentiation without decreasing TGF-b receptor expression.

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Section: D Effects Of Tgf-b1mentioning

confidence: 99%

Response of Fibroblasts to Transforming Growth Factor-β1 on Two-Dimensional and in Three-Dimensional Hyaluronan Hydrogels

Chen

Thibeault

2012

Tissue Engineering Part A

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“…29 To generate mechanical stress in the collagen matrix, after polymerization of the lattice mixture, the lattices remained attached to the culture well and were cultured in FCM for 24 hours to allow the development of mechanical tension. At 24 hours, the lattice was fixed with 4% paraformaldehyde, then completely detached with a pipette tip.…”

Section: Immunodetection Of Myofibroblast Differentiation In Mechanicmentioning

confidence: 99%

Profibrotic Phenotype of Conjunctival Fibroblasts from Mucous Membrane Pemphigoid

Saw

Schmidt

Offiah

et al. 2011

The American Journal of Pathology

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Ocular mucous membrane pemphigoid is an immunobullous disease in which excessive conjunctival fibrosis causes blindness, and the pathogenesis of scarring is incompletely understood. To establish whether profibrotic fibroblasts with an altered phenotype exist in ocular mucous membrane pemphigoid, we compared the functional characteristics of pemphigoid conjunctival fibroblasts to normal conjunctival fibroblasts with respect to cell division; migration; collagen contraction; matrix metalloproteinase, secretion of collagen and chemokines; and myofibroblast differentiation. We found that pemphigoid fibroblasts showed increased cell division (P ‫؍‬ 0.01), increased migration in serum-free medium (72 ؎ 18 migrated cells versus 33 ؎ 11, P ‫؍‬ 0.04), increased collagen contraction in the presence of 10 ng/ml tumor necrosis factor-␣, increased collagen type I secretion (P ‫؍‬ 0.03), increased secretion of matrix metalloproteinase-3 (P ‫؍‬ 0.03), and increased secretion of eotaxin in response to interleukin-13 (P ‫؍‬ 0.04). Differences between pemphigoid and normal conjunctival fibroblasts with respect to collagen contraction and MMP secretion in the presence of interleukin-13 were also observed. Together, these findings indicate that pemphigoid conjunctival fibroblasts have a profibrotic phenotype that is maintained in vitro. No differences between pemphigoid fibroblasts obtained from acutely inflamed versus clinically uninflamed conjunctiva were observed. Developing effective antifibrotic therapies will require understanding of the mechanisms that both induce and maintain the profibrotic phenotype.

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“…Biological properties of CTGF include involvement in cell proliferation, adhesion, chemotaxis, and extracellular matrix production in a variety of cell types such as fibroblasts, mesangial cells, endothelial cells, and chondrocytes (15,43,60). Since CTGF appears to be a critical factor in the development of tubulointerstitial fibrosis, acting as a downstream mediator of TGF-␤ 1 signaling (20), and is differentially regulated during EMT (17,18,24,37), understanding the mechanisms controlling CTGF expression is of paramount importance in defining the role of this molecule in renal disease.…”

mentioning

confidence: 99%

Hypoxic induction ofCtgfis directly mediated by Hif-1

Higgins

Biju²,

Akai³

et al. 2004

American Journal of Physiology-Renal Physiology

255

196

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CTGF plays a significant role in the development of renal fibrosis by mediating the fibrotic effects of transforming growth factor (TGF)-β1and has been shown to be hypoxia inducible in human breast cancer cells. It has been suggested that hypoxia is an important underlying cause for the development of renal fibrosis through the modulation of profibrotic genes. One of the key mediators of the cell's response to lowered oxygen environments is hypoxia-inducible-factor-1 (HIF-1), a basic helix-loop-helix transcription factor, which enables cells to adapt to hypoxia by regulating the expression of genes involved in increasing oxygen availability ( VEGF, erythropoietin) and enhancing glucose uptake and metabolism ( Glut-1, PGK). In this paper, we have used primary tubular epithelial cell cultures from a tetracycline-inducible- Hif- 1α knockout murine model to further elucidate the role of Hif-1 in the hypoxic-induction of Ctgf expression. We show that hypoxia response elements present upstream of Ctgf enable direct interaction of Hif-1 transcription factor with the Ctgf promoter, resulting in increased transcription of Ctgf mRNA. Cells deficient in Hif- 1α were incapable of inducing Ctgf mRNA in response to hypoxia, suggesting an absolute requirement of Hif-1. Furthermore, the observed Hif-1-mediated hypoxic stimulation of Ctgf expression was found to occur independently of TGF-β1signaling. Our findings have important implications for a number of fibrotic disorders in which hypoxia, CTGF, and TGF-β1are involved, including renal, dermal, hepatic, and pulmonary fibrosis.

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Involvement of CTGF in TGF-β1–Stimulation of Myofibroblast Differentiation and Collagen Matrix Contraction in the Presence of Mechanical Stress

Cited by 124 publications

References 38 publications

Response of Fibroblasts to Transforming Growth Factor-β1 on Two-Dimensional and in Three-Dimensional Hyaluronan Hydrogels

Response of Fibroblasts to Transforming Growth Factor-β1 on Two-Dimensional and in Three-Dimensional Hyaluronan Hydrogels

Profibrotic Phenotype of Conjunctival Fibroblasts from Mucous Membrane Pemphigoid

Hypoxic induction ofCtgfis directly mediated by Hif-1

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