May 2007
Volume 48, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2007
Inhibition of Corneal Epithelial Cell Production of Fibrotic Mediator TGF-beta2 by Basement Membrane-Like Extracellular Matrix
Author Affiliations & Notes
  • M. Fini
    University of Miami Miller School of Medicine, Miami, Florida
    Bascom Palmer Eye Institute,
  • A. J. LaGier
    University of Miami Miller School of Medicine, Miami, Florida
    Bascom Palmer Eye Institute,
  • S. Yoo
    University of Miami Miller School of Medicine, Miami, Florida
    Bascom Palmer Eye Institute,
  • E. C. Alfonso
    University of Miami Miller School of Medicine, Miami, Florida
    Bascom Palmer Eye Institute,
  • S. A. Meiners
    Pharmacology, Robert Wood Johnson Medical School, UMDMJ, Piscataway, New Jersey
  • V. Lemmon
    University of Miami Miller School of Medicine, Miami, Florida
    Miami Project to Cure Paralysis,
  • Footnotes
    Commercial Relationships M. Fini, None; A.J. LaGier, None; S. Yoo, None; E.C. Alfonso, None; S.A. Meiners, None; V. Lemmon, None.
  • Footnotes
    Support NIH Grant EY09828 and EY014801; Walter G. Ross Foundation; Research to Prevent Blindness, Inc.; RPB Senior Scientific Investigator Award; Walter G. Ross Foundation
Investigative Ophthalmology & Visual Science May 2007, Vol.48, 3499. doi:
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    • Get Citation

      M. Fini, A. J. LaGier, S. Yoo, E. C. Alfonso, S. A. Meiners, V. Lemmon; Inhibition of Corneal Epithelial Cell Production of Fibrotic Mediator TGF-beta2 by Basement Membrane-Like Extracellular Matrix. Invest. Ophthalmol. Vis. Sci. 2007;48(13):3499.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose:: Transforming Growth Factor-beta2 (TGF-beta2) is a major epithelial mediator of fibrotic marker expression. Production of TGF-beta2 protein is reduced by plating on matrigel, a basement membrane-like extracellular matrix extract. The goal of the current study was to further understand the mechanism of matrigel regulation.

Methods:: We used a human epithelial cell culture model for these studies. Chemical panel screening was done in multi-well plate format using cells transfected with a beta-lactamase reporter gene construct driven by the TGF-beta2 promoter.

Results:: TGF-beta2 production is inhibited by matrigel at the level of mRNA accumulation and activity of the TGF-beta2 gene transcriptional promoter. The effects of matrigel cannot be explained by growth factor contaminants, reduction in heparin sulfate proteoglycans has no effect, and pure forms of the major ECM components laminin and collagen IV can not alone reproduce the effect. Experiments also failed to implicate inhibition of a constitutive TGF-beta2 autocrine feedback loop. Thus we went on to consider hypotheses that incorporated a requirement for complexity. Matrigel caused a reduction of ezrin, a member of the ezrin/ radixin/ moesin (ERM) family, which plays a role in establishing polarity of epithelial cells in tissues through the Rho signaling pathway and the actin cytoskelton. A set of know chemical compounds was screened for the capacity to inhibit TGF-beta2 promoter activity, yielding two hits - one an inhibiter of actin polymerization and another an inhibitor of the PDGF receptor tyrosine kinase.

Conclusions:: These findings indicate that matrigel inhibits TGF-beta2 gene expression and point to a mechanism requiring the complexity of matrigel composition and structure. Ezrin and the actin cytoskelton, Rho kinase, and the PDGF receptor tyrosine kinase are implicated in this mechanism. Chemical compounds that affect this pathway might be used as pharmacologic modulators to improve the quality of wound healing and refractive surgical outcomes in the cornea.

Keywords: cornea: epithelium • growth factors/growth factor receptors 
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