Regular Article
Peptides Encoded by Exon 6 of VEGF Inhibit Endothelial Cell Biological Responses and Angiogenesis Induced by VEGF

https://doi.org/10.1006/bbrc.2001.4761Get rights and content

Abstract

VEGF induces pathological angiogenesis and is an important target for the development of novel antiangiogenic molecules. In this study, we tested synthetic peptides based on the sequence of VEGF189 for their ability to inhibit VEGF receptor binding and biological responses. We identified 12-amino acid peptides derived from exon 6 that inhibited VEGF binding to HUVECs, VEGF-stimulated ERK activation, and prostacyclin production. These peptides inhibited VEGF-induced mitogenesis, migration, and VEGF-dependent survival of endothelial cells, but caused no increase in apoptosis in the absence of VEGF. Exon 6-encoded peptides also caused a marked inhibition of VEGF-induced angiogenesis in vitro. Studies of effects of peptides on cross-linking of VEGF to its receptors and on binding of VEGF to porcine aortic endothelial cells expressing either KDR or neuropilin-1 showed that exon 6-encoded peptides effectively blocked the interaction of VEGF with both receptors. Exon 6-derived peptides caused release of bFGF from endothelial cells but inhibited bFGF-dependent ERK activation, cell proliferation and angiogenesis. Our findings indicate that VEGF exon 6-encoded peptides inhibit VEGF-induced angiogenesis, at least in part through inhibition of VEGF binding to KDR. In addition, exon 6-encoded peptides are also effective inhibitors of bFGF-mediated angiogenesis.

References (40)

  • C. Wheeler-Jones et al.

    Vascular endothelial growth factor stimulates prostacyclin production and activation of cytosolic phospholipase A2 in endothelial cells via p42/p44 mitogen-activated protein kinase

    FEBS Lett.

    (1997)
  • H. Abedi et al.

    Vascular endothelial growth factor stimulates tyrosine phosphorylation and recruitment to new focal adhesions of focal adhesion kinase and paxillin in endothelial cells

    J. Biol. Chem.

    (1997)
  • B.A. Keyt et al.

    Identification of vascular endothelial growth factor determinants for binding KDR and FLT-1 receptors. Generation of receptor-selective VEGF variants by site-directed mutagenesis

    J. Biol. Chem.

    (1996)
  • S. Soker et al.

    Characterization of novel vascular endothelial growth factor (VEGF) receptors on tumor cells that bind VEGF165 via its exon 7-encoded domain

    J. Biol. Chem.

    (1996)
  • F. Jonca et al.

    Cell release of bioactive fibroblast growth factor 2 by exon 6-encoded sequence of vascular endothelial growth factor

    J. Biol. Chem.

    (1997)
  • J. Waltenberger et al.

    Different signal transduction properties of KDR and Flt1, two receptors for vascular endothelial growth factor

    J. Biol. Chem.

    (1994)
  • S. Soker et al.

    Inhibition of vascular endothelial growth factor (VEGF)-induced endothelial cell proliferation by a peptide corresponding to the exon 7-encoded domain of VEGF165

    J. Biol. Chem.

    (1997)
  • R. Volk et al.

    The role of syndecan cytoplasmic domain in basic fibroblast growth factor-dependent signal transduction

    J. Biol. Chem.

    (1999)
  • G. Neufeld et al.

    Vascular endothelial growth factor (VEGF) and its receptors

    FASEB J.

    (1999)
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    Abbreviations used: bFGF, basic fibroblast growth factor; DSS, disuccinimidyl suberate; ERK1,2, extracellular signal-regulated kinases 1 and 2; HUVEC, human umbilical vein endothelial cells; NP-1, neuropilin-1; PAE/KDR, porcine aortic endothelial cells expressing KDR; PAE/NP-1, porcine aortic endothelial cells expressing NP-1; PGI2, prostacyclin; SDS–PAGE, sodium dodecyl sulphate polyacrylamide gel electrophoresis; VEGF, vascular endothelial growth factor.

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    To whom correspondence may be addressed. Haiyan Jia: Fax: 020-7679-6212. E-mail: [email protected].. Ian Zachary: Fax: 020-7679-6212. E-mail: [email protected]

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