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TGF-ß2–Induced Cell Surface Tissue Transglutaminase Increases Adhesion and Migration of RPE Cells on Fibronectin through the Gelatin-Binding Domain

From the Department of Ophthalmology, Ludwig-Maximilians-University, Munich, Germany.

PURPOSE. Migration and adhesion of dislocated retinal pigment epithelial (RPE) cells to a fibronectin-rich extracellular matrix is an initial step in proliferative vitreoretinopathy (PVR). In the present study, the functional role of cell surface tissue transglutaminase (tTG) in adhesion and migration of RPE cells on fibronectin (Fn) and collagen type I (Col I) after stimulation with TGF-ß2 was investigated.

METHODS. Cultured human RPE cells were treated with 1.0 ng/mL TGF-ß2 for 24 hours. Cell surface tTG expression was determined by cell fraction analysis. Attachment on Col I, full-length Fn, and its 45-kDa gelatin-binding and 110-kDa cell-binding fragment was measured with an MTT assay. Migration of RPE cells was measured by a Boyden chamber assay, and cell spreading was determined. Experiments were performed in the presence or absence of anti-tTG antibodies and anti-integrin 5 and ß1 antibodies.

RESULTS. TGF-ß2 markedly induced expression of cell-surface tTG on RPE cells and increased attachment and migration on Fn and Col I. Blocking cell surface tTG inhibited attachment, migration, and spreading on Fn and its 45-kDa gelatin-binding fragment, whereas no effect was seen on Col I and the 110-kDa cell-binding Fn fragment. In contrast, blocking of integrin 5 and ß1 suppressed adhesion and migration on full-length Fn and the 110-kDa Fn fragment.

CONCLUSIONS. These data demonstrate that TGF-ß2 increases expression of cell surface tTG, which in turn strengthens adhesion, migration, and spreading of RPE cells on Fn through the 45-kDa gelatin-binding Fn fragment. At the onset of PVR, this mechanism may help RPE cells to attach and migrate on Fn-containing matrices.

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