Functional Ephrin-B2 Expression for Promotive Interaction Between Arterial and Venous Vessels in Postnatal Neovascularization

Background—Ephrin-B2, one of the transmembrane ligands, is a genetic marker of arterial endothelial cells (ECs) at embryonic stages and is essential for cardiovascular development, but its roles in ischemic cardiovascular disease are not well understood. In this study, we focused on the function of ephrin-B2 in postnatal neovascularization. Methods and Results—We found that ephrin-B2 is exclusively expressed and significantly upregulated in the arterial vasculature after the initial angiogenic responses in tissue ischemia. Upregulation of ephrin-B2 is also observed in EC cordlike formation in vitro. Interestingly, ephrin-B2 expression on ECs was enhanced by promotive angiogenic growth factors, such as vascular endothelial growth factor, basic fibroblast growth factor, and hepatocyte growth factor, whereas it was attenuated by angiopoietin-1, a factor for blood vessel maturation. Moreover, an ephrin-B2–rich environment was shown to induce neovascularization mainly through venous angiogenesis in an in vivo cornea micropocket assay. Conclusions—Our study indicates that the ephrin-B2 ligand is likely to have functional expression on angiogenic arterial ECs and induce a subsequent promotive effect on venous vessels during postnatal neovascularization.

[1]  Chad A. Cowan,et al.  Ephrin-B2 reverse signaling is required for axon pathfinding and cardiac valve formation but not early vascular development. , 2004, Developmental biology.

[2]  Guoyao Wu,et al.  Eph B4 Receptor Signaling Mediates Endothelial Cell Migration and Proliferation via the Phosphatidylinositol 3-Kinase Pathway* , 2002, The Journal of Biological Chemistry.

[3]  G. Yancopoulos,et al.  EphB ligand, ephrinB2, suppresses the VEGF‐ and angiopoietin‐1‐induced Ras/mitogen‐activated protein kinase pathway in venous endothelial cells , 2002, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[4]  B. Weinstein,et al.  sonic hedgehog and vascular endothelial growth factor act upstream of the Notch pathway during arterial endothelial differentiation. , 2002, Developmental cell.

[5]  David J. Anderson,et al.  Sensory Nerves Determine the Pattern of Arterial Differentiation and Blood Vessel Branching in the Skin , 2002, Cell.

[6]  X. Q. Zhang,et al.  Stromal cells expressing ephrin-B2 promote the growth and sprouting of ephrin-B2(+) endothelial cells. , 2001, Blood.

[7]  G. Yancopoulos,et al.  Ephrin-B2 selectively marks arterial vessels and neovascularization sites in the adult, with expression in both endothelial and smooth-muscle cells. , 2001, Developmental biology.

[8]  G Garcia-Cardena,et al.  Expression of ephrinB2 identifies a stable genetic difference between arterial and venous vascular smooth muscle as well as endothelial cells, and marks subsets of microvessels at sites of adult neovascularization. , 2001, Developmental biology.

[9]  R. D'Amato,et al.  Genetic heterogeneity of angiogenesis in mice , 2000, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[10]  A. Brändli,et al.  The receptor tyrosine kinase EphB4 and ephrin-B ligands restrict angiogenic growth of embryonic veins in Xenopus laevis. , 2000, Development.

[11]  I. Daar,et al.  Fibroblast Growth Factor Receptor-Mediated Rescue of x-Ephrin B1-Induced Cell Dissociation in XenopusEmbryos , 2000, Molecular and Cellular Biology.

[12]  Thomas N. Sato,et al.  Leakage-resistant blood vessels in mice transgenically overexpressing angiopoietin-1. , 1999, Science.

[13]  D. Anderson,et al.  Symmetrical mutant phenotypes of the receptor EphB4 and its specific transmembrane ligand ephrin-B2 in cardiovascular development. , 1999, Molecular cell.

[14]  Qiling Xu,et al.  Eph receptors and ephrins restrict cell intermingling and communication , 1999, Nature.

[15]  Y. Taniyama,et al.  Therapeutic angiogenesis induced by human recombinant hepatocyte growth factor in rabbit hind limb ischemia model as cytokine supplement therapy. , 1999, Hypertension.

[16]  R. Klein,et al.  Eph receptors and ephrins: effectors of morphogenesis. , 1999, Development.

[17]  G. Yancopoulos,et al.  Growth factors acting via endothelial cell-specific receptor tyrosine kinases: VEGFs, angiopoietins, and ephrins in vascular development. , 1999, Genes & development.

[18]  A. Lane,et al.  Surface densities of ephrin‐B1 determine EphB1‐coupled activation of cell attachment through αvβ3 and α5β1 integrins , 1999 .

[19]  F. Diella,et al.  Roles of ephrinB ligands and EphB receptors in cardiovascular development: demarcation of arterial/venous domains, vascular morphogenesis, and sprouting angiogenesis. , 1999, Genes & development.

[20]  R. Huganir,et al.  PDZ Proteins Bind, Cluster, and Synaptically Colocalize with Eph Receptors and Their Ephrin Ligands , 1998, Neuron.

[21]  G. Yancopoulos,et al.  Critical role of the TIE2 endothelial cell receptor in the development of definitive hematopoiesis. , 1998, Immunity.

[22]  J. Isner,et al.  Tie2 receptor ligands, angiopoietin-1 and angiopoietin-2, modulate VEGF-induced postnatal neovascularization. , 1998, Circulation research.

[23]  J. Isner,et al.  Mouse model of angiogenesis. , 1998, The American journal of pathology.

[24]  David J. Anderson,et al.  Molecular Distinction and Angiogenic Interaction between Embryonic Arteries and Veins Revealed by ephrin-B2 and Its Receptor Eph-B4 , 1998, Cell.

[25]  J. Folkman,et al.  Vasculogenesis, Angiogenesis, and Growth Factors: Ephrins Enter the Fray at the Border , 1998, Cell.

[26]  A. Lane,et al.  Eph receptors discriminate specific ligand oligomers to determine alternative signaling complexes, attachment, and assembly responses. , 1998, Genes & development.

[27]  P. Huang,et al.  Nitric oxide synthase modulates angiogenesis in response to tissue ischemia. , 1998, The Journal of clinical investigation.

[28]  Douglas Hanahan,et al.  Signaling Vascular Morphogenesis and Maintenance , 1997, Science.

[29]  Thomas N. Sato,et al.  Angiopoietin-2, a natural antagonist for Tie2 that disrupts in vivo angiogenesis. , 1997, Science.

[30]  W. Risau,et al.  Mechanisms of angiogenesis , 1997, Nature.

[31]  E. Pasquale,et al.  Tyrosine Phosphorylation of Transmembrane Ligands for Eph Receptors , 1997, Science.

[32]  David J. Anderson,et al.  Eph Family Transmembrane Ligands Can Mediate Repulsive Guidance of Trunk Neural Crest Migration and Motor Axon Outgrowth , 1997, Neuron.

[33]  J. Ware,et al.  Angiogenesis in ischemic heart disease , 1997, Nature Medicine.

[34]  Pamela F. Jones,et al.  Requisite Role of Angiopoietin-1, a Ligand for the TIE2 Receptor, during Embryonic Angiogenesis , 1996, Cell.

[35]  T. Pawson,et al.  Bidirectional signalling through the EPH-family receptor Nuk and its transmembrane ligands , 1996, Nature.

[36]  Takayuki Asahara,et al.  Clinical evidence of angiogenesis after arterial gene transfer of phVEGF165 in patient with ischaemic limb , 1996, The Lancet.

[37]  J. Folkman,et al.  A model of angiogenesis in the mouse cornea. , 1996, Investigative ophthalmology & visual science.

[38]  Lieve Moons,et al.  Abnormal blood vessel development and lethality in embryos lacking a single VEGF allele , 1996, Nature.

[39]  Kenneth J. Hillan,et al.  Heterozygous embryonic lethality induced by targeted inactivation of the VEGF gene , 1996, Nature.

[40]  J. Rossant,et al.  The receptor tyrosine kinase TIE is required for integrity and survival of vascular endothelial cells. , 1995, The EMBO journal.

[41]  J. Flanagan,et al.  ELF-2, a new member of the Eph ligand family, is segmentally expressed in mouse embryos in the region of the hindbrain and newly forming somites , 1995, Molecular and cellular biology.

[42]  Janet Rossant,et al.  Failure of blood-island formation and vasculogenesis in Flk-1-deficient mice , 1995, Nature.

[43]  Thomas N. Sato,et al.  Distinct roles of the receptor tyrosine kinases Tie-1 and Tie-2 in blood vessel formation , 1995, Nature.

[44]  J. Rossant,et al.  Role of the Flt-1 receptor tyrosine kinase in regulating the assembly of vascular endothelium , 1995, Nature.

[45]  K. Alitalo,et al.  Vascularization of the mouse embryo: A study of flk‐1, tek, tie, and vascular endothelial growth factor expression during development , 1995, Developmental dynamics : an official publication of the American Association of Anatomists.

[46]  A. Goddard,et al.  Molecular cloning of a ligand for the EPH-related receptor protein-tyrosine kinase Htk. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[47]  W Grossman,et al.  Basic fibroblast growth factor improves myocardial function in chronically ischemic porcine hearts. , 1994, The Journal of clinical investigation.

[48]  E. Brogi,et al.  Therapeutic angiogenesis. A single intraarterial bolus of vascular endothelial growth factor augments revascularization in a rabbit ischemic hind limb model. , 1994, The Journal of clinical investigation.

[49]  J G Flanagan,et al.  The ephrins and Eph receptors in neural development. , 1998, Annual review of neuroscience.

[50]  J. Folkman.  Angiogenesis in cancer, vascular, rheumatoid and other disease , 1995, Nature Medicine.