September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
The role of FAK in fibrotic matrix contraction by dedifferentiated Müller cells
Author Affiliations & Notes
  • Rintaro Tsukahara
    Ophthalmology, Tokyo Med Univ, Ibaraki Med Ctr, Inashikigun, Ibaraki, Japan
  • Kazuhiko Umazume
    Ophthalmology, Tokyo Med Univ, Shinjuku, Tokyo, Japan
  • Naoyuki Yamakawa
    Ophthalmology, Tokyo Med Univ, Shinjuku, Tokyo, Japan
  • Takuya Iwasaki
    Ophthalmology, Tokyo Med Univ, Ibaraki Med Ctr, Inashikigun, Ibaraki, Japan
  • Henry J Kaplan
    Ophthalmology and Visual Sciences, University of Louisville, Louisville, Kentucky, United States
  • Hiroshi Goto
    Ophthalmology, Tokyo Med Univ, Shinjuku, Tokyo, Japan
  • Shigeo Tamiya
    Ophthalmology and Visual Sciences, University of Louisville, Louisville, Kentucky, United States
    Biochemistry and Molecular Biology, University of Louisville, Louisville, Kentucky, United States
  • Footnotes
    Commercial Relationships   Rintaro Tsukahara, None; Kazuhiko Umazume, None; Naoyuki Yamakawa, None; Takuya Iwasaki, None; Henry Kaplan, None; Hiroshi Goto, None; Shigeo Tamiya, None
  • Footnotes
    Support  DoD/USAMRAA DMRDP-ARATDA DM090475; Unrestricted institutional grant from Research to Prevent Blindness, NY, NY
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 4195. doi:
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    • Get Citation

      Rintaro Tsukahara, Kazuhiko Umazume, Naoyuki Yamakawa, Takuya Iwasaki, Henry J Kaplan, Hiroshi Goto, Shigeo Tamiya; The role of FAK in fibrotic matrix contraction by dedifferentiated Müller cells. Invest. Ophthalmol. Vis. Sci. 2016;57(12):4195.

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

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Abstract

Purpose : We have previously reported the inhibitory effect of dasatinib on fibrotic matrix contraction using both in vitro and in vivo models (Umazume K, et al IOVS 2013, Tsukahara R, et al EER 2015). Dasatinib was developed as a dual inhibitor of SFK and ABL tyrosine kinases but has also been reported to inhibit other tyrosine kinases including focal adhesion kinase (FAK). In this study, we examined the effect of FAK inhibition on matrix contraction by dedifferentiated Müller cells, which has been implicated in several ocular fibrotic complications.

Methods : Müller cells were isolated from porcine eyes using a papain/DNase kit, and used between passages 3-6. Cells were cultured for 3 days in 25% vitreous fluid supplemented DMEM in the presence or absence of dasatinib or PF573228 (FAK inhibitor). Expression of active (phosphorylated) and total FAK protein expression was examined by Western blot analyses. A type I collagen matrix contraction assays were used to examine matrix contraction.

Results : Dedifferentiated Müller cells expressed active FAK. PF573228 treatment significantly reduced active FAK expression and inhibited matrix contraction. When added early (on day 0) prior to cell clustering and pseudo-ERM formation, the inhibitory effect on matrix contraction was comparable to dasatinib. On the other hand, addition of PF573228 after cell clustering (on day 2) resulted in a suppressive effect that was still significant but much diminished in comparison to dasatinib.

Conclusions : FAK is involved in matrix contraction by differentiated Müller cells, with the main role early on prior to and/or during cell clustering and ERM formation.

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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