RGD-dependent binding of TP508 to integrin αvβ3 mediates cell adhesion and induction of nitric oxide

Dmitry N. Derkach; Subhagya A. Wadekar; Kim B. Perkins; Emma Rousseau; Catherine M. Dreiza; Joyce Cheung-Flynn; Heidi C. Ramos; Tatiana P. Ugarova; Michael R. Sheller

doi:10.1160/TH09-07-0447

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 2010; 104(01): 172-182
DOI: 10.1160/TH09-07-0447

New Technologies, Diagnostic Tools and Drugs

Schattauer GmbH

RGD-dependent binding of TP508 to integrin α_vβ₃ mediates cell adhesion and induction of nitric oxide

Dmitry N. Derkach^*

¹Capstone Therapeutics, Tempe, Arizona, USA

,

Subhagya A. Wadekar^*

¹Capstone Therapeutics, Tempe, Arizona, USA

,

Kim B. Perkins

¹Capstone Therapeutics, Tempe, Arizona, USA

,

Emma Rousseau

¹Capstone Therapeutics, Tempe, Arizona, USA

,

Catherine M. Dreiza

¹Capstone Therapeutics, Tempe, Arizona, USA

²Applied Sciences and Mathematics, Arizona State University, Tempe, Arizona, USA

,

Joyce Cheung-Flynn

¹Capstone Therapeutics, Tempe, Arizona, USA

,

Heidi C. Ramos

¹Capstone Therapeutics, Tempe, Arizona, USA

,

Tatiana P. Ugarova

³Center for Metabolic Biology, Arizona State University, Tempe, Arizona, USA

,

Michael R. Sheller

¹Capstone Therapeutics, Tempe, Arizona, USA

› Institutsangaben

Abstract

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Summary

P508, a 23-amino acid RGD-containing synthetic peptide representing residues 508 to 530 of human prothrombin, mitigates the effects of endothelial dysfunction in ischaemic reperfusion injury. The objective of this study was to investigate whether TP508 binds to members of the integrin family of transmembrane receptors leading to nitric oxide synthesis. Immobilised TP508 supported adhesion of endothelial cells and α_vβ₃-expressing human embryonic kidney cells in a dose- and RGD-dependent manner. Soluble TP508 also inhibited cell adhesion to immobilised fibrinogen. The involvement of α_vβ₃ was verified with function-blocking antibodies and surface plasmon resonance studies. Adhesion of the cells to immobilised TP508 resulted in an induction of phosphorylated FAK and ERK1/2. In endothelial cells, TP508 treatment resulted in an induction of nitric oxide that could be inhibited by LM609, an α_vβ₃-specific, function-blocking monoclonal antibody. Finally, TP508 treatment of isolated rat aorta segments enhanced carbachol-induced vasorelaxation. These results suggest that TP508 elicits a potentially therapeutic effect through an RGD-dependent interaction with integrin α_vβ₃.

Keywords

TP508 - eNOS - nitric - oxide - endothelial - dysfunction - integrin

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Referenzen

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