Inhibition of angiogenesis by growth factor receptor bound protein 2-Src homology 2 domain bound antagonists.
Growth factor receptor bound protein 2 (Grb2) is an intracellular adaptor protein that participates in the signal transduction cascades of several angiogenic factors, including hepatocyte growth factor, basic fibroblast growth factor, and vascular endothelial growth factor. We described previously the potent blockade of hepatocyte growth factor-stimulated cell motility, matrix invasion, and epithelial tubulogenesis by synthetic Grb2-Src homology 2 (SH2) domain binding antagonists. Here, we show that these binding antagonists block basic morphogenetic events required for angiogenesis, including hepatocyte growth factor-, vascular endothelial growth factor-, and basic fibroblast growth factor-stimulated endothelial cell proliferation and migration, as well as phorbol 12-myristate 13-acetate-stimulated endothelial cell migration and matrix invasion. The Grb2-SH2 domain binding antagonists also impair angiogenesis in vitro, as shown by the inhibition of cord formation by macrovascular endothelial cells on Matrigel. We further show that a representative compound inhibits angiogenesis in vivo as measured using a chick chorioallantoic membrane assay. These results suggest that Grb2 is an important mediator of key proangiogenic events, with potential application to pathologic conditions where neovascularization contributes to disease progression. In particular, the well-characterized role of Grb2 in signaling cell cycle progression together with our present findings suggests that Grb2-SH2 domain binding antagonists have the potential to act as anticancer drugs that target both tumor and vascular cell compartments.
Cellular and Molecular Biology, National Cancer Institute, Building 10, Room 2B47, 9000 Rockville Pike, Bethesda, MD 20892-1501, USA.
Jesus V
JV
Soriano
Ningfei
N
Liu
Yang
Y
Gao
Zhu-Jun
ZJ
Yao
Toshio
T
Ishibashi
Charles
C
Underhill
Terrence R
TR
Burke
Donald P
DP
Bottaro
D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism / Q000502:physiology*; D000818:Animals; D002460:Cell Line; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D002478:Cells, Cultured; D002642:Chick Embryo; D003094:Collagen / Q000494:pharmacology; D018450:Disease Progression; D004338:Drug Combinations; D004730:Endothelium, Vascular / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism; D051380:GRB2 Adaptor Protein; D006133:Growth Substances / Q000378:metabolism*; D006801:Humans; D007797:Laminin / Q000494:pharmacology; D008956:Models, Chemical; D009389:Neovascularization, Pathologic; D011485:Protein Binding; D011509:Proteoglycans / Q000494:pharmacology; D015398:Signal Transduction; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D018909:src Homology Domains
D048868:Adaptor Proteins, Signal Transducing; D004338:Drug Combinations; D051380:GRB2 Adaptor Protein; C495948:GRB2 protein, human; D006133:Growth Substances; D007797:Laminin; D011509:Proteoglycans; D016222:Fibroblast Growth Factor 2; C059485:matrigel; D003094:Collagen; D013755:Tetradecanoylphorbol Acetate