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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. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
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.
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.
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.
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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