Selective inhibition of vascular endothelial growth factor receptor-2 (VEGXR-2) identifies a central role for VEGFR-2 in human aortic endothelial cell responses to VEGF

被引:34
作者
Endo, A [1 ]
Fukuhara, S [1 ]
Masuda, M [1 ]
Ohmori, T [1 ]
Mochizuki, N [1 ]
机构
[1] Natl Cardiovasc Ctr, Inst Res, Dept Struct Anal, Suita, Osaka 5658565, Japan
关键词
VEGF receptor; inhibitor; angiogenesis;
D O I
10.1081/RRS-120025567
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Vascular endothelial growth factor receptors (VEGFR) are considered essential for angiogenesis. The VEGFR-family proteins consist of VEGFR-1/Flt-1, VEGFR-2/ KDR/Flk- 1, and VEGFR-3/Flt-4. Among these, VEGFR-2 is thought to be principally responsible for angiogenesis. However, the precise role of VEGFRs1-3 in endothelial cell biology and angiogenesis remains unclear due in part to the lack of VEGFR-specific inhibitors. We used the newly described, highly selective anilinoquinazoline inhibitor of VEGFR-2 tyrosine kinase, ZM323881 (5-[[7-(benzyloxy) quinazolin-4- yl]amino]-4-fluoro-2-methylphenol), to explore the role of VEGFR-2 in endothelial cell function. Consistent with its reported effects on VEGFR-2 [IC(50) < 2 nM], ZM323881 inhibited activation of VEGFR-2, but not of VEGFR-1, epidermal growth factor receptor (EGFR), platelet-derived growth factor receptor (PDGFR), or hepatocyte growth factor (HGF) receptor., We studied the effects of VEGF on human aortic endothelial cells (HAECs), which express VEGFR-1 and VEGFR-2, but not VEGFR-3, in the absence or presence of ZM323881. Inhibition of VEGFR-2 blocked activation of extracellular regulated-kinase, p38, Akt, and endothelial nitric oxide synthetase (eNOS) by VEGF, but did not inhibit p38 activation by the VEGFR-1-specific ligand, placental growth factor (PIGF). Inhibition of VEGFR-2 also perturbed VEGF-induced membrane extension, cell migration, and tube formation by HAECs. Vascular endothelial growth factor receptor-2 inhibition also reversed VEGF-stimulated phosphorylation of CrkII and its Src homology 2 (SH2)-binding protein p130(Cas) which are known to play a pivotal role in regulating endothelial cell migration. Inhibition of VEGFR-2 thus blocked all VEGF-induced endothelial cellular responses tested, supporting that the catalytic activity of VEGFR-2 is critical for VEGF signaling and/or that VEGFR-2 may function in a heterodimer with VEGFR-1 in human vascular endothelial cells.
引用
收藏
页码:239 / 254
页数:16
相关论文
共 39 条
[1]   Role of PIGF in the intra- and intermolecular cross talk between the VEGF receptors Flt1 and Flk1 [J].
Autiero, M ;
Waltenberger, J ;
Communi, D ;
Kranz, A ;
Moons, L ;
Lambrechts, D ;
Kroll, J ;
Plaisance, S ;
De Mol, M ;
Bono, F ;
Kliche, S ;
Fellbrich, G ;
Ballmer-Hofer, K ;
Maglione, D ;
Mayr-Beyrle, U ;
Dewerchin, M ;
Dombrowski, S ;
Stanimirovic, D ;
Van Hummelen, P ;
Dehio, C ;
Hicklin, DJ ;
Persico, G ;
Herbert, JM ;
Communi, D ;
Shibuya, M ;
Collen, D ;
Conway, EM ;
Carmeliet, P .
NATURE MEDICINE, 2003, 9 (07) :936-943
[2]   Ras and Rho GTPases: A family reunion [J].
Bar-Sagi, D ;
Hall, A .
CELL, 2000, 103 (02) :227-238
[3]   Vascular endothelial growth factor effect on endothelial cell proliferation, migration, and platelet-activating factor synthesis is Flk-1-dependent [J].
Bernatchez, PN ;
Soker, S ;
Sirois, MG .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (43) :31047-31054
[4]   Rho GTPases and their effector proteins [J].
Bishop, AL ;
Hall, A .
BIOCHEMICAL JOURNAL, 2000, 348 (02) :241-255
[5]   Synergism between vascular endothelial growth factor and placental growth factor contributes to angiogenesis and plasma extravasation in pathological conditions [J].
Carmeliet, P ;
Moons, L ;
Luttun, A ;
Vincenti, V ;
Compernolle, V ;
De Mol, M ;
Wu, Y ;
Bon, F ;
Devy, L ;
Beck, H ;
Scholz, D ;
Acker, T ;
DiPalma, T ;
Dewerchin, M ;
Noel, A ;
Stalmans, I ;
Barra, A ;
Blacher, S ;
Vandendriessche, T ;
Ponten, A ;
Eriksson, U ;
Plate, KH ;
Foidart, JM ;
Schaper, W ;
Charnock-Jones, DS ;
Hicklin, DJ ;
Herbert, JM ;
Collen, D ;
Persico, MG .
NATURE MEDICINE, 2001, 7 (05) :575-583
[6]   Mammalian MAP kinase signalling cascades [J].
Chang, LF ;
Karin, M .
NATURE, 2001, 410 (6824) :37-40
[7]   Sphingosine 1-phosphate induces membrane ruffling and increases motility of human umbilical vein endothelial cells via vascular endothelial growth factor receptor and CrkII [J].
Endo, A ;
Nagashima, KI ;
Kurose, H ;
Mochizuki, S ;
Matsuda, M ;
Mochizuki, N .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2002, 277 (26) :23747-23754
[8]   The biology of vascular endothelial growth factor [J].
Ferrara, N ;
DavisSmyth, T .
ENDOCRINE REVIEWS, 1997, 18 (01) :4-25
[9]   ROLE OF THE FLT-1 RECEPTOR TYROSINE KINASE IN REGULATING THE ASSEMBLY OF VASCULAR ENDOTHELIUM [J].
FONG, GH ;
ROSSANT, J ;
GERTSENSTEIN, M ;
BREITMAN, ML .
NATURE, 1995, 376 (6535) :66-70
[10]   Vascular endothelial growth factor regulates endothelial cell survival through the phosphatidylinositol 3′-kinase Akt signal transduction pathway -: Requirement for Flk-1/KDR activation [J].
Gerber, HP ;
McMurtrey, A ;
Kowalski, J ;
Yan, MH ;
Keyt, BA ;
Dixit, V ;
Ferrara, N .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1998, 273 (46) :30336-30343