1. Vascular endothelial growth factor selectively targets boronated dendrimers to tumor vasculature.
- Author
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Backer MV, Gaynutdinov TI, Patel V, Bandyopadhyaya AK, Thirumamagal BT, Tjarks W, Barth RF, Claffey K, and Backer JM
- Subjects
- Animals, Boron Compounds chemistry, Boron Neutron Capture Therapy, Cell Proliferation drug effects, Female, Humans, Mammary Neoplasms, Experimental metabolism, Mice, Mice, Inbred BALB C, Neovascularization, Pathologic metabolism, Polyamines chemistry, Receptors, Vascular Endothelial Growth Factor metabolism, Recombinant Fusion Proteins administration & dosage, Recombinant Fusion Proteins pharmacology, Tumor Cells, Cultured, Vascular Endothelial Growth Factor A administration & dosage, Boron Compounds pharmacokinetics, Endothelium, Vascular drug effects, Mammary Neoplasms, Experimental blood supply, Neovascularization, Pathologic drug therapy, Polyamines pharmacokinetics, Vascular Endothelial Growth Factor A pharmacology
- Abstract
Tumor neovasculature is a potential but, until very recently, unexplored target for boron neutron capture therapy (BNCT) of cancer. In the present report, we describe the construction of a vascular endothelial growth factor (VEGF)-containing bioconjugate that potentially could be used to target up-regulated VEGF receptors (VEGFR), which are overexpressed on tumor neovasculature. A fifth-generation polyamidoamine dendrimer containing 128 reactive amino groups was reacted with 105 to 110 decaborate molecules to produce a macromolecule with 1,050 to 1,100 boron atoms per dendrimer. This was conjugated to thiol groups of VEGF at a 4:1 molar ratio using the heterobifunctional reagent sulfo-LC-SPDP. In addition, the boronated dendrimer was tagged with a near-IR Cy5 dye to allow for near-IR fluorescent imaging of the bioconjugate in vitro and in vivo. As would be predicted, the resulting VEGF-BD/Cy5 bioconjugate was not cytotoxic to HEK293 cells engineered to express 2.5 x 10(6) VEGFR-2 per cell. Furthermore, it showed binding and activation of VEGFR-2 comparable with that of native VEGF. Internalization of VEGF-BD/Cy5 by PAE cells expressing 2.5 x 10(5) VEGFR-2 per cell was inhibited by excess VEGF, indicating a VEGFR-2-mediated mechanism of uptake. Near-IR fluorescent imaging of 4T1 mouse breast carcinoma revealed selective accumulation of VEGF-BD/Cy5, but not BD/Cy5, particularly at the tumor periphery where angiogenesis was most active. Accumulation of VEGF-BD/Cy5 in 4T1 breast carcinoma was diminished in mice pretreated with a toxin-VEGF fusion protein that selectively killed VEGFR-2-overexpressing endothelial cells. Our data lay the groundwork for future studies using the VEGF-BD/Cy5 bioconjugate as a targeting agent for BNCT of tumor neovasculature.
- Published
- 2005
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