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Journals The FASEB Journal November 2009

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Extracellular cadherin repeat domains EC1 and EC5 of T-cadherin are essential for its ability to stimulate angiogenic behavior of endothelial cells

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T-cadherin (T-cad) promotes survival, proliferation, and migration of endothelial cells and induces angiogenesis. We aimed to identify domains of T-cad functionally relevant to its effects on endothelial cell behavior. To specifically target the functional properties of the 5 cadherin repeat domains (EC1–EC5) of T-cad, endothelial cells were transduced with lentivectors containing specific T-cad-domain-deletion mutant constructs ( I, II, III, IV, V). Empty (E) lentivector-transduced cells served as control. Similarly to overexpression of native T-cad, cells expressing II, III, or IV displayed elevated levels of p-Akt and p-GSK3β and increased proliferation rates (for II, III) vs. E. I- and V-transduced cells exhibited reduced levels of p-Akt and p-GSK3β and retarded growth rates vs. E. Stimulatory effects of native T-cad overexpression on Akt and GSK3β phosphorylation were dose dependently inhibited by coexpression of I or V. Subsequent functional analyses compared only I-, II-, and V-mutant constructs with E as a negative control. Unlike II cells, I and V cells failed to exhibit homophilic ligation and deadhesion responses on a substratum of T-cad protein. In the wound assay, migration was increased for II cells but impaired for I and V cells. In endothelial cell-spheroid assay, angiogenic sprouting was augmented for II cells but inhibited for I and V cells. We conclude that EC1 and EC5 domains of T-cad are essential for its proangiogenic effects. I and V constructs may serve as dominant-negative mutants and as potential tools targeting excessive angiogenesis.—Joshi, M. B., Kyriakakis, E., Pfaff, D., Rupp, K., Philippova, M., Erne, P., Resink, T. J. Extracellular cadherin repeat domains EC1 and EC5 of T-cadherin are essential for its ability to stimulate angiogenic behavior of endothelial cells.

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Article Details
Joshi, Manjunath B.; Kyriakakis, Emmanouil; Pfaff, Dennis; Rupp, Katharina; Philippova, Maria; Erne, Paul; Resink, Therese J.
The FASEB Journal , Volume 23 (11): 4011
Fed of American Socs for Experimental Biology – Nov 1, 2009
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