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Mechanisms of Tumor Cell Interaction With Vascular Cells

B. Felding-Habermann, T.J. Kunicki, R. Billetta,^* S. Ferrone,^** G.H. Ryu,^*** Z.M. Ruggeri

^* University of California, San Diego, CA
^** New York Medical College, Valhalla, NY
^*** Bureau of Medical and Radiation Health, Seoul, Korea

Hematogenous metastasis requires attachment of tumor cells to the vessel wall during blood flow. Using an in vitro perfusion system, we found that tumor cell arrest involved interaction between tumor cells and activated platelets via ß₃ integrins and cross-linking plasma proteins. Recognition between ß₃ integrins and their ligands involves the Arg-Gly-Asp (RGD) ligand motif. Therefore, we analyzed critical flanking sequences surrounding the RGD site and the conformational constraints of the RGD domain that are crucial in determining the ligand specificity for tumor cell integrin _vß₃ and platelet integrin _IIbß₃. This information will be important for the development of agents that inhibit interactions between tumor cells and platelets.

Another receptor of potential importance in adhesion of tumor cells to vascular cells is intracellular adhesion molecule-1. We found a clear correlation between expression of this molecule and tumor progression in melanoma patients, indicating a role for the molecule in adhesion of melanoma cells during the metastatic cascade. Because of the clinical significance of the expression of integrin _vß₃ and intracellular adhesion molecule-1 in melanoma lesions, these receptors are targets for future studies on melanoma cell arrest and extravasation. For this purpose, we have developed an experimental system that enables us to analyze both tumor cell attachment to the endothelium and transendothelial migration of tumor cells during blood flow.

Critical parameters in this in vitro system are stimulation of the endothelium and its presentation of a shear-resistant three-dimensional cushion that enable us to analyze transmigrated cells. We will use three-dimensional reconstruction of confocal images acquired during the perfusion to detect and quantify attached and penetrated cells. This technique will enable us to study vascular cells and their receptors and ligands that are involved in adhesive and invasive tumor cell interaction with the vessel wall and that affect the rate of hematogenous tumor metastasis.

PUBLICATIONS

Celikel, R., Madhusudan, Varughese, K.I., Shima, M., Yoshioka, A., Ware, J., Ruggeri, Z.M. Crystal structure of NMC-4 Fab anti-von Willebrand factor A1 domain. Blood Cells Mol. Dis. 23:123, 1997.

Federici, A.B., Mannucci, P.M., Stabile, F., Canciani, M.T., Di Rocco, N., Miyata, S., Ware, J., Ruggeri, Z.M. A type 2B von Willebrand disease mutation (Ile⁵⁴⁶Val) associated with an unusual phenotype. Thromb. Haemost. 78:1132, 1997.

Hayashi, T., Ware, J., Niiya, K., Sakuragawa, N. Isolated recombinant domain of von Willebrand factor displaying increased sensitivity to ristocetin Am. J. Hematol. 52:248, 1996.

Kunicki, T.J, Annis, D.S, Felding-Habermann, B. Molecular determinants of Arg-Gly-Asp ligand specificity for ß₃ integrins. J. Biol. Chem. 272:4103, 1997.

Lanza, P., Felding-Habermann, B., Ruggeri, Z.M, Zanetti, M., Billetta, R. RGD conformationally-constrained in an antibody loop interacts selectively with the _vß₃ integrin on tumor cells. Blood Cells Mol. Dis. 23:230, 1997.

Natali, P.G., Hamby, C.V., Felding-Habermann, B., Liang, B., Nicotra, M.R., Di Filippo, F., Giannarelli, D., Temponi, M., Ferrone, S. Clinical significance of _vß₃ integrin and intercellular adhesion molecule-1 expression in cutaneous malignant melanoma lesions. Cancer Res. 57:1554, 1997.

Pareti, F.I., Cattaneo, M., Carpinelli, L., Zighetti, M.L., Bressi, C., Mannucci, P.M., Ruggeri, Z.M. Evaluation of the abnormal platelet function in von Willebrand disease by the blood filtration test. Thromb. Haemost. 75:460, 1996.

Ware, J., Hashimoto, Y., Zieger, B., Russell, A. Controlling elements of platelet glycoprotein Ib expression. C. R. Acad. Sci. 319:811, 1996.

Zieger, B., Hashimoto, Y., Ware., J. Alternative expression of platelet glycoprotein Ibß mRNA from an adjacent 5´ gene with an imperfect polyadenylation signal sequence. J. Clin. Invest. 99:520, 1997.

Zieger, B., Ware, J. Cloning and deduced amino acid sequence of human nicotinamide nucleotide transhydrogenase. DNA Seq. 7:369, 1997.