Dasatinib affects focal adhesion and myosin regulation to inhibit matrix contraction by Muller cells

作者：Tsukahara Rintaro; Umazume Kazuhiko; Yamakawa Naoyuki; McDonald Kevin; Kaplan Henry J; Tamiya Shigeo^*

来源：Experimental Eye Research, 2015, 139: 90-96.

DOI：10.1016/j.exer.2015.07.019

• Muller cells
• Epiretinal membrane
• Contraction
• Dasatinib
• Focal adhesion
• Stress fibers

Epiretinal membrane (ERM) contraction is associated with a variety of ocular diseases that cause macular dysfunction. Trans-differentiated Muller cells have been identified in ERMs, and have been implicated to be involved in the contractile process. In this study, we tested the effect of dasatinib, an FDA-approved tyrosine kinase inhibitor, on matrix contraction caused by Muller cells, and examined molecular mechanism of action. Type I collagen matrix contraction assays were used to examine the effect of drugs on matrix contraction by trans-differentiated Muller cells. Fluophore-conjugated phalloidin was used for the detection of actin cytoskeleton, and Western-blot analyses were carried out to examine protein expression and phosphorylation status. Dasatinib inhibited collagen matrix contraction by transdifferentiated Muller cells that was associated with decreased cell spreading and reduction of actomyosin stress fibers. Concomitantly, dasatinib-treated Muller cells had reduced phosphorylation of Src family kinase, paxillin, as well as myosin II light chain. Specific inhibitors of Rho/ROCK and myosin II confirmed the critical role played by this pathway in Muller cell contraction. Our data demonstrate that dasatinib significantly reduced matrix contraction by Muller cells via inhibition of focal adhesion, as well as actomyosin contraction.

• 出版日期2015-10