Antagonistic Anti-urokinase Plasminogen Activator Receptor (uPAR) Antibodies Significantly Inhibit uPAR-mediated Cellular Signaling and Migration

Department of Pharmaceutical Chemistry, University of California, San Francisco, California 94158-2517, USA.
Journal of Biological Chemistry (Impact Factor: 4.57). 08/2010; 285(35):26878-88. DOI: 10.1074/jbc.M109.077677
Source: PubMed


Interactions between urokinase plasminogen activator receptor (uPAR) and its various ligands regulate tumor growth, invasion, and metastasis. Antibodies that bind specific uPAR epitopes may disrupt these interactions, thereby inhibiting these processes. Using a highly diverse and naïve human fragment of the antigen binding (Fab) phage display library, we identified 12 unique human Fabs that bind uPAR. Two of these antibodies compete against urokinase plasminogen activator (uPA) for uPAR binding, whereas a third competes with beta1 integrins for uPAR binding. These competitive antibodies inhibit uPAR-dependent cell signaling and invasion in the non-small cell lung cancer cell line, H1299. Additionally, the integrin-blocking antibody abrogates uPAR/beta1 integrin-mediated H1299 cell adhesion to fibronectin and vitronectin. This antibody and one of the uPAR/uPA antagonist antibodies shows a significant combined effect in inhibiting cell invasion through Matrigel/Collagen I or Collagen I matrices. Our results indicate that these antagonistic antibodies have potential for the detection and treatment of uPAR-expressing tumors.

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    • "Cell Line Studies The clonogenic survival and matrigel invasion assays were conducted as previously described 19, 20. For the qPCR analysis RNA was prepared from each cell line (~ 2 x 106 cells/cell line) using an RNEasy kit (Qiagen). "
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