Stimulation of endothelial cell proliferation by FGF-2 in the presence of fibrinogen requires αvβ3

We have shown previously that fibrin(ogen) binding potentiates the capacity of fibroblast growth factor 2 (FGF-2) to stimulate endothelial cell (EC) proliferation. We have now investigated the receptor requirement for EC proliferation by fibrinogen-bound FGF-2. ECs were cultured with 25 ng/mL FGF-2 with or without 10 microg/mL fibrinogen, and proliferation was measured as (3)H-thymidine incorporation. Proliferation was increased 2.4 +/- 0.5-fold over medium alone with FGF-2 and increased significantly more to 4.0 +/- 0.7-fold with fibrinogen and FGF-2 (P < .005). Addition of 7E3 or LM609, antibodies to alpha(v)beta(3), inhibited EC proliferation with fibrinogen-bound FGF-2 by 80% +/- 8% (P < .001) or 67% +/- 14% (P < .002), respectively, to levels significantly less than that observed with FGF-2 alone (P < .001). Neither LM609 nor 7E3 exhibited any inhibition of activity with FGF-2 alone. Peptide GRGDS caused dose-dependent inhibition of proliferation by fibrinogen-bound FGF-2 of 31% +/- 8%, 45% +/- 9%, and 68% +/- 11% at 0.25, 0.5, and 1 mM, respectively. Coimmunoprecipitation and immunofluorescence studies demonstrated a direct specific association between alpha(v)beta(3) and FGF receptor 1 (FGFR1) in ECs and fibroblasts when exposed to both FGF-2 and fibrinogen but not with vitronectin. We conclude that fibrinogen binding of FGF-2 enhances EC proliferation through the coordinated effects of colocalized alpha(v)beta(3) and FGFR1.

[1]  A. Sahni,et al.  Identification of a binding site on human FGF-2 for fibrinogen. , 2004, Blood.

[2]  A. Sahni,et al.  FGF‐2 but not FGF‐1 binds fibrin and supports prolonged endothelial cell growth , 2003, Journal of thrombosis and haemostasis : JTH.

[3]  B. Lowell,et al.  Adipose tissue mass can be regulated through the vasculature , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[4]  Salman Rahman,et al.  Novel Vascular Endothelial Growth Factor Binding Domains of Fibronectin Enhance Vascular Endothelial Growth Factor Biological Activity , 2002, Circulation research.

[5]  P. Dell’Era,et al.  Biological activity of substrate-bound basic fibroblast growth factor (FGF2): recruitment of FGF receptor-1 in endothelial cell adhesion contacts , 2002, Oncogene.

[6]  B. Eliceiri.  This Review is part of a thematic series on Integrins, which includes the following articles: Integrins and the Myocardium Functional Consequences of Integrin Gene Mutations in Mice Integrins in Vascular Development Integrin and Growth Factor Receptor Crosstalk , 2001 .

[7]  F. Isik,et al.  Integrin activation is required for VEGF and FGF receptor protein presence on human microvascular endothelial cells , 2001, Molecular and Cellular Biochemistry.

[8]  A. Sahni,et al.  Vascular endothelial growth factor binds to fibrinogen and fibrin and stimulates endothelial cell proliferation. , 2000, Blood.

[9]  J. Schlessinger,et al.  Crystal structure of a ternary FGF-FGFR-heparin complex reveals a dual role for heparin in FGFR binding and dimerization. , 2000, Molecular cell.

[10]  A. Sahni,et al.  Fibrinogen and Fibrin Protect Fibroblast Growth Factor-2 from Proteolytic Degradation , 2000, Thrombosis and Haemostasis.

[11]  S. Hubbard,et al.  Structural Basis for FGF Receptor Dimerization and Activation , 1999, Cell.

[12]  A. Sahni,et al.  Potentiation of Endothelial Cell Proliferation by Fibrin(ogen)-bound Fibroblast Growth Factor-2* , 1999, The Journal of Biological Chemistry.

[13]  L. Silengo,et al.  Integrins induce activation of EGF receptor: role in MAP kinase induction and adhesion‐dependent cell survival , 1998, The EMBO journal.

[14]  A. Sahni,et al.  Binding of Basic Fibroblast Growth Factor to Fibrinogen and Fibrin* , 1998, The Journal of Biological Chemistry.

[15]  G. Garcı́a-Cardeña,et al.  The synergistic activity of alphavbeta3 integrin and PDGF receptor increases cell migration. , 1998, Journal of cell science.

[16]  Marco Presta,et al.  αvβ3 Integrin Mediates the Cell-adhesive Capacity and Biological Activity of Basic Fibroblast Growth Factor (FGF-2) in Cultured Endothelial Cells , 1997 .

[17]  E. Ruoslahti,et al.  αvβ3 integrin associates with activated insulin and PDGFβ receptors and potentiates the biological activity of PDGF , 1997 .

[18]  D. Birnbaum,et al.  Murine FGF-12 and FGF-13: expression in embryonic nervous system, connective tissue and heart , 1997, Mechanisms of Development.

[19]  M. Pepper,et al.  Manipulating angiogenesis. From basic science to the bedside. , 1997, Arteriosclerosis, thrombosis, and vascular biology.

[20]  E. Plow,et al.  Fibrinogen Is a Ligand for Integrin α5β1 on Endothelial Cells* , 1997, The Journal of Biological Chemistry.

[21]  K. Yamada,et al.  Integrins can collaborate with growth factors for phosphorylation of receptor tyrosine kinases and MAP kinase activation: roles of integrin aggregation and occupancy of receptors , 1996, The Journal of cell biology.

[22]  K. Rubin,et al.  Stimulation of beta1 integrins on fibroblasts induces PDGF independent tyrosine phosphorylation of PDGF beta-receptors , 1996, The Journal of cell biology.

[23]  I. Lax,et al.  Regulation of growth factor activation by proteoglycans: What is the role of the low affinity receptors? , 1995, Cell.

[24]  D E Ingber,et al.  Convergence of integrin and growth factor receptor signaling pathways within the focal adhesion complex. , 1995, Molecular biology of the cell.

[25]  K. Okada,et al.  Effects of fibrin on the secretion of plasminogen activator inhibitor-1 from endothelial cells and on protein kinase C. , 1995, Life sciences.

[26]  G. Gillespie,et al.  Vascular endothelial growth factor in human glioma cell lines: induced secretion by EGF, PDGF-BB, and bFGF. , 1995, Journal of Neurosurgery.

[27]  M. Presta,et al.  Transcriptional and posttranscriptional regulation of urokinase‐type plasminogen activator expression in endothelial cells by basic fibroblast growth factor , 1995, Journal of cellular physiology.

[28]  N. Ling,et al.  Identification of soluble forms of the fibroblast growth factor receptor in blood. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[29]  D. Rifkin,et al.  Basic fibroblast growth factor modulates integrin expression in microvascular endothelial cells. , 1993, Molecular biology of the cell.

[30]  L. Languino,et al.  Fibrinogen mediates leukocyte adhesion to vascular endothelium through an ICAM-1-dependent pathway , 1993, Cell.

[31]  P. Hall,et al.  Overview of the fibroblast growth factor and receptor families: complexity, functional diversity, and implications for future cardiovascular research. , 1993, Cardiovascular research.

[32]  A. Logan,et al.  Angiogenesis , 1993, The Lancet.

[33]  M. Siegman,et al.  Elimination of smooth muscle cells in experimental restenosis: targeting of fibroblast growth factor receptors. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[34]  E. Dejana,et al.  Human endothelial cells express integrin receptors on the luminal aspect of their membrane. , 1992, Blood.

[35]  C. Francis,et al.  Endothelial cell spreading on fibrin requires fibrinopeptide B cleavage and amino acid residues 15-42 of the beta chain. , 1992, The Journal of clinical investigation.

[36]  D. Rifkin,et al.  Expression of the urokinase receptor in vascular endothelial cells is stimulated by basic fibroblast growth factor , 1991, The Journal of cell biology.

[37]  M. Jaye,et al.  Cloning and expression of two distinct high‐affinity receptors cross‐reacting with acidic and basic fibroblast growth factors. , 1990, The EMBO journal.

[38]  L. Orci,et al.  Transforming growth factor-beta 1 modulates basic fibroblast growth factor-induced proteolytic and angiogenic properties of endothelial cells in vitro , 1990, The Journal of cell biology.

[39]  Y. Shimada,et al.  Apoptosis of vascular endothelial cells by fibroblast growth factor deprivation. , 1990, Biochemical and biophysical research communications.

[40]  J W Smith,et al.  Integrin (alpha v beta 3)-ligand interaction. Identification of a heterodimeric RGD binding site on the vitronectin receptor. , 1990, The Journal of biological chemistry.

[41]  D. Cheresh,et al.  Recognition of distinct adhesive sites on fibrinogen by related integrins on platelets and endothelial cells , 1989, Cell.

[42]  R. Mesa-Tejada,et al.  Immunohistochemical characterization of fibrin(ogen)-related antigens in human tissues using monoclonal antibodies. , 1989, Laboratory investigation; a journal of technical methods and pathology.

[43]  D. Rifkin,et al.  Alpha 2-macroglobulin is a binding protein for basic fibroblast growth factor. , 1989, The Journal of biological chemistry.

[44]  R. Mesa-Tejada,et al.  Identification and Distribution of Fibrinogen, Fibrin, and Fibrin(ogen) Products in Atherosclerosis: Use of Monoclonal Antibodies , 1989, Arteriosclerosis.

[45]  D. Rifkin,et al.  Interaction of heparin with human basic fibroblast growth factor: Protection of the angiogenic protein from proteolytic degradation by a glycosaminoglycan , 1989, Journal of cellular physiology.

[46]  M. Presta,et al.  Purification from a human hepatoma cell line of a basic fibroblast growth factor-like molecule that stimulates capillary endothelial cell plasminogen activator production, DNA synthesis, and migration , 1986, Molecular and cellular biology.

[47]  M. Presta,et al.  Purification of a factor from human placenta that stimulates capillary endothelial cell protease production, DNA synthesis, and migration. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[48]  V. Terranova,et al.  Human endothelial cells are chemotactic to endothelial cell growth factor and heparin , 1985, The Journal of cell biology.

[49]  J. Denekamp,et al.  Endothelial proliferation in tumours and normal tissues: continuous labelling studies. , 1984, British Journal of Cancer.

[50]  E. Plow,et al.  Fibrinogen is a ligand for integrin alpha5beta1 on endothelial cells. , 1997, The Journal of biological chemistry.

[51]  D. Rifkin,et al.  Fibroblast growth factors as angiogenesis factors: new insights into their mechanism of action. , 1997, EXS.

[52]  P. Dell’Era,et al.  alphavbeta3 integrin mediates the cell-adhesive capacity and biological activity of basic fibroblast growth factor (FGF-2) in cultured endothelial cells. , 1997, Molecular biology of the cell.

[53]  E. Ruoslahti,et al.  Alphavbeta3 integrin associates with activated insulin and PDGFbeta receptors and potentiates the biological activity of PDGF. , 1997, The EMBO journal.

[54]  D. Moscatelli,et al.  The FGF family of growth factors and oncogenes. , 1992, Advances in cancer research.

[55]  A. Bini.  Identification and distribution of fibrinogen, fibrin and fibrin (ogen) degradation products in atherosclerosis , 1989 .

[56]  A. Mantovani,et al.  Interaction between fibrinogen and cultured endothelial cells. Induction of migration and specific binding. , 1985, The Journal of clinical investigation.