September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
PYK2 plays a role in matrix contraction by RPE-derived cells
Author Affiliations & Notes
  • Shigeo Tamiya
    Ophthalmology and Visual Sciences, University of Louisville, Louisville, Kentucky, United States
  • Kevin McDonald
    Ophthalmology and Visual Sciences, University of Louisville, Louisville, Kentucky, United States
  • Henry J Kaplan
    Ophthalmology and Visual Sciences, University of Louisville, Louisville, Kentucky, United States
  • Footnotes
    Commercial Relationships   Shigeo Tamiya, None; Kevin McDonald, None; Henry Kaplan, None
  • Footnotes
    Support  DoD/USAMRAA BAA W81XWH-15-1-0298; Research to Prevent Blindness, New York, NY
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 246. doi:
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    • Get Citation

      Shigeo Tamiya, Kevin McDonald, Henry J Kaplan; PYK2 plays a role in matrix contraction by RPE-derived cells. Invest. Ophthalmol. Vis. Sci. 2016;57(12):246.

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      © 2017 Association for Research in Vision and Ophthalmology.

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Abstract

Purpose : Contractile fibroblastic/myofibroblastic cells derived from retinal pigment epithelial (RPE) cells have been implicated in the fibrotic complications of proliferative vitreoretinopathy. We have previously demonstrated that matrix contraction by RPE-derived cells can be inhibited by dasatinib, an FDA-approved cancer medication that inhibits multiple tyrosine kinases. The purpose of this project was to identify molecular targets affected by dasatinib that contribute to matrix contraction.

Methods : Primary cultured porcine RPE cells were used between passages 3 to 5. Cells were cultured for 3 days in 25% vitreous fluid supplemented DMEM in the presence or absence of tyrosine kinase inhibitors for the final 24 hours. Western blot analyses were used to determine the phosphorylation status of phospho-tyrosine proteins. A type I collagen contraction assay was utilized to examine matrix contraction by cells.

Results : Dasatinib reduced tyrosine phosphorylation of multiple proteins. Interestingly, while FAK auto-phosphorylation on Tyr397 was unaffected, auto-phosphorylation on Tyr402 of PYK2, also known as FAK2, was significantly reduced. In agreement with the Western blot data, PF431396, a dual inhibitor of PYK2 and FAK, significantly reduced matrix contraction while PF573228, an inhibitor of FAK but not PYK2, was without effect.

Conclusions : The inhibitory effect of dasatinib on matrix contraction by RPE-derived cells is, at least in part, due to the prevention of PYK2 activation, which plays a role in the contractile process.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

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