Abstract
Background
During vein graft adaptation to the arterial circulation, vascular endothelial growth
factor (VEGF) A expression transiently increases before becoming downregulated; however,
the role of VEGF-A in venous remodeling is not clear. In addition, although VEGF-A
stimulates angiogenesis and determines arterial identity in nascent arterial endothelial
cells (EC), the role of VEGF-A in regulating identity in adult venous EC is also not
clear.
Materials and methods
EC, wild type (EphB4+/+) or heterozygous knockout (EphB4+/−), were stimulated with
VEGF-A (0–100 ng/mL) and examined with quantitative polymerase chain reaction and
western blotting.
Results
VEGF-A (100 ng/mL) inhibited expression of EphB4 and stimulated expression of delta-like
ligand 4 (dll4) but did not stimulate either notch or EphrinB2 expression in adult
venous EC. Pretreatment with VEGF receptor 2–neutralizing antibody abolished VEGF-stimulated
downregulation of EphB4 but not the upregulation of dll4. Pretreatment with PD98059
or wortmannin showed that VEGF-A downregulation of EphB4 and upregulation of dll4
are mitogen-activated protein kinase kinase and extracellular signal–regulated kinase
dependent but phosphatidylinositol 3 kinase–Akt independent. Compared with VEGF-induced
EphB4 downregulation and dll4 upregulation in control EC, reduced EphB4 signaling
in EphB4+/− EC showed even further downregulation of EphB4 and upregulation of dll4.
Conclusions
Despite the genetic programming of arterial and venous EC fate, VEGF-A can repress
venous identity in adult venous EC without induction of arterial identity. These changes
in adult EC in vitro recapitulate the changes in identity described during vein graft adaptation to the
arterial environment in vivo.
Keywords
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References
- Biochemical (functional) adaptation of “arterialized” vein grafts.Ann Surg. 1986; 203: 339
- The natural history of endothelial structure and function in arterialized vein grafts.J Vasc Surg. 1986; 3: 204
- Mechanisms of vein graft adaptation to the arterial circulation: insights into the neointimal algorithm and management strategies.Circ J. 2010; 74: 1501
- Control of blood vessel identity: from embryo to adult.Ann Vasc Dis. 2008; 1: 28
- Eph-B4 prevents venous adaptive remodeling in the adult arterial environment.J Exp Med. 2011; 208: 561
- Molecular distinction and angiogenic interaction between embryonic arteries and veins revealed by ephrin-B2 and its receptor Eph-B4.Cell. 1998; 93: 741
- Ephrin-B2 selectively marks arterial vessels and neovascularization sites in the adult, with expression in both endothelial and smooth-muscle cells.Dev Biol. 2001; 230: 151
- Venous identity is lost but arterial identity is not gained during vein graft adaptation.Arterioscler Thromb Vasc Biol. 2007; 27: 1562
- Abnormal blood vessel development and lethality in embryos lacking a single VEGF allele.Nature. 1996; 380: 435
- Heterozygous embryonic lethality induced by targeted inactivation of the VEGF gene.Nature. 1996; 380: 439
- Functional ephrin-B2 expression for promotive interaction between arterial and venous vessels in postnatal neovascularization.Circulation. 2005; 111: 2210
- Suppression of Notch signalling by the COUP-TFII transcription factor regulates vein identity.Nature. 2005; 435: 98
- Specification of arterial, venous, and lymphatic endothelial cells during embryonic development.Histol Histopathol. 2010; 25: 637
- Sonic hedgehog and vascular endothelial growth factor act up-stream of the Notch pathway during arterial endothelial differentiation.Dev Cell. 2002; 3: 127
- ERK1/2-Akt1 crosstalk regulates arteriogenesis in mice and zebrafish.J Clin Invest. 2010; 120: 1217
- Reduction of vein graft intimal hyperplasia and preservation of endothelium-dependent relaxation by topical vascular endothelial growth factor.J Vasc Surg. 1998; 27: 167
- Akt1/protein kinase Bα is critical for ischemic and VEGF-mediated angiogenesis.J Clin Invest. 2005; 115: 2119
- EphB ligand, ephrinB2, suppresses the VEGF- and angiopoietin-1-induced Ras/mitogen-activated protein kinase pathway in venous endothelial cells.FASEB J. 2002; 16: 1126
- The role of the vascular endothelial growth factor-delta-like 4 ligand/Notch4-ephrin B2 cascade in tumor vessel remodeling and endothelial cell functions.Cancer Res. 2006; 66: 8501
- Shear stress increases expression of the arterial endothelial marker ephrinB2 in murine ES cells via the VEGF-Notch signaling pathways.Arterioscler Thromb Vasc Biol. 2009; 29: 2125
- Gene expression changes evoked in a venous segment exposed to arterial flow.J Vasc Surg. 2006; 44: 863
- An overview of the Notch signalling pathway.Semin Cell Dev Biol. 2003; 14: 113
- Notch signaling is required for arterial-venous differentiation during embryonic vascular development.Development. 2001; 128: 3675
- Vascular patterning defects associated with expression of activated Notch4 in embryonic endothelium.Proc Natl Acad Sci USA. 2001; 98: 5643
- High affinity VEGF binding and developmental expression suggests FIk-l as a major regulator of angiogenesis and vasculogenesis.Cell. 1993; 72: 835
- Flt-1 lacking the tyrosine kinase domain is sufficient for normal development and angiogenesis in mice.Proc Natl Acad Sci U S A. 1998; 95: 9349
- VEGF receptor 2 endocytic trafficking regulates arterial morphogenesis.Dev Cell. 2010; 18: 713
- Artery/vein specification is governed by opposing phosphatidylinositol-3 kinase and MAP kinase/ERK signaling.Curr Biol. 2006; 16: 1366
- Symmetrical mutant phenotypes of the receptor EphB4 and its specific transmembrane ligand ephrin-B2 in cardiovascular development.Mol Cell. 1999; 4: 403
- Up-regulation of the Notch ligand delta-like 4 inhibits VEGF-induced endothelial cell function.Blood. 2006; 107: 931
- Arterial-venous specification during development.Circ Res. 2009; 104: 576
- Notch-mediated restoration of regenerative potential to aged muscle.Science. 2003; 302: 1575
- Age-related Notch-4 quiescence is associated with altered wall remodeling during vein graft adaptation.J Surg Res. 2011; 171: e149
- Vascular endothelial growth factor receptor-2: structure, function, intracellular signalling and therapeutic inhibition.Cell Signal. 2007; 19: 2003
- Differential expression of neuropilin-1 and neuropilin-2 in arteries and veins.Mech Dev. 2001; 109: 115
- Neuropilin-2 is a receptor for the vascular endothelial growth factor (VEGF) forms VEGF-145 and VEGF-165.J Biol Chem. 2000; 275: 29922
- Targeting neuropilin-1 to inhibit VEGF signaling in cancer: comparison of therapeutic approaches.PLoS Comput Biol. 2006; 2: e180
- Neuropilin-2: a new molecular target for antiangiogenic and antitumor strategies.J Natl Cancer Inst. 2008; 100: 81
- Neuropilins in neoplasms: expression, regulation, and function.Exp Cell Res. 2006; 312: 584
- MAPping out arteries and veins.Sci STKE. 2006; 355: pe39
- Shear stress activates p60src-Ras-MAPK signaling pathways in vascular endothelial cells.Arterioscler Thromb Vasc Biol. 1998; 18: 227
- MAPKs (ERK1/2, p38) and AKT can be phosphorylated by shear stress independently of platelet endothelial cell adhesion molecule-1 (CD31) in vascular endothelial cells.J Biol Chem. 2005; 280: 11185
- The range of adaptation by collateral vessels after femoral artery occlusion.Circ Res. 2006; 99: 656
- Evidence of a role for phosphatidylinositol 3-kinase activation in the blocking of apoptosis by polyomavirus middle T antigen.J Virol. 1998; 72: 3221
Article info
Publication history
Published online: February 04, 2013
Accepted:
January 3,
2013
Received in revised form:
December 18,
2012
Received:
November 7,
2012
Identification
Copyright
Published by Elsevier Inc.
