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(American Journal of Pathology. 1998;153:381-394.)
© 1998 American Society for Investigative Pathology


Regular Articles

Vascular Endothelial Growth Factor-C (VEGF-C/VEGF-2) Promotes Angiogenesis in the Setting of Tissue Ischemia

Bernhard Witzenbichler* , Takayuki Asahara* , Toyoaki Murohara* , Marcy Silver* , Ioakim Spyridopoulos* , Meredith Magner* , Nicole Principe* , Marianne Kearney* , Jing-Shan Hu{dagger} and Jeffrey M. Isner*

From the Departments of Medicine (Cardiology) and Biomedical Research,* St. Elizabeth's Medical Center of Boston, Tufts University School of Medicine, Boston, Massachusetts, and the Department of Protein Therapeutics,{dagger} Human Genome Sciences, Inc., Rockville, Maryland

Recently, vascular endothelial growth factor-C (VEGF-C or VEGF-2) was described as a specific ligand for the endothelial receptor tyrosine kinases VEGFR-2 and VEGFR-3. In vivo data, limited to constitutive overexpression in transgenic mice, have been interpreted as evidence that the growth-promoting effects of VEGF-C are restricted to development of the lymphatic vasculature. The current studies were designed to test the hypothesis that constitutive expression of VEGF-C in adult animals promotes angiogenesis. In vitro, VEGF-C exhibited a dose-dependent mitogenic and chemotactic effect on endothelial cells, particularly for microvascular endothelial cells (72% and 95% potency, respectively, compared with VEGF-A/VEGF-1). VEGF-C stimulated release of nitric oxide from endothelial cells and increased vascular permeability in the Miles assay; the latter effect was attenuated by pretreatment with the nitric oxide synthase inhibitor N{omega}-nitro-L-arginine methyl ester. Both VEGFR-2 and VEGFR-3 receptors were shown to be expressed in human saphenous vein and internal mammary artery. The potential for VEGF-C to promote angiogenesis in vivo was then tested in a rabbit ischemic hindlimb model. Ten days after ligation of the external iliac artery, VEGF-C was administered as naked plasmid DNA (pcVEGF-C; 500 µg) from the polymer coating of an angioplasty balloon (n = 8 each) or as recombinant human protein (rhVEGF-C; 500 µg) by direct intra-arterial infusion. Physiological and anatomical assessments of angiogenesis 30 days later showed evidence of therapeutic angiogenesis for both pcVEGF-C and rhVEGF-C. Hindlimb blood pressure ratio (ischemic/normal) after pcVEGF-C increased to 0.83 ± 0.03 after pcVEGF-C versus 0.59 ± 0.04 (P < 0.005) in pGSVLacZ controls and to 0.76 ± 0.04 after rhVEGF-C versus 0.58 ± 0.03 (P < 0.01) in control rabbits receiving rabbit serum albumin. Doppler-derived iliac flow reserve was 2.7 ± 0.1 versus 2.0 ± 0.2 (P < 0.05) for pcVEGF-C versus LacZ controls and 2.9 ± 0.3 versus 2.1 ± 0.2 (P < 0.05) for rhVEGF-C versus albumin controls. Neovascularity was documented by angiography in vivo (angiographic scores: 0.85 ± 0.05 versus 0.51 ± 0.02 (P < 0.001) for plasmid DNA and 0.74 ± 0.08 versus 0.53 ± 0.03 (P < 0.05) for protein), and capillary density (per mm2) was measured at necropsy (252 ± 12 versus 183 ± 10 (P < 0.005) for plasmid DNA and 229 ± 20 versus 164 ± 20 (P < 0.05) for protein). In contrast to the results of gene targeting experiments, constitutive expression of VEGF-C in adult animals promotes angiogenesis in the setting of limb ischemia. VEGF-C and its receptors thus constitute an apparently redundant pathway for postnatal angiogenesis and may represent an alternative to VEGF-A for strategies of therapeutic angiogenesis in patients with limb and/or myocardial ischemia.





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A. Saaristo, T. A. Partanen, J. Arola, L. Jussila, M. Hytonen, A. Makitie, S. Vento, A. Kaipainen, H. Malmberg, and K. Alitalo
Vascular Endothelial Growth Factor-C and Its Receptor VEGFR-3 in the Nasal Mucosa and in Nasopharyngeal Tumors
Am. J. Pathol., July 1, 2000; 157(1): 7 - 14.
[Abstract] [Full Text] [PDF]


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Arterioscler. Thromb. Vasc. Bio.Home page
I. Zachary, A. Mathur, S. Yla-Herttuala, and J. Martin
Vascular Protection : A Novel Nonangiogenic Cardiovascular Role for Vascular Endothelial Growth Factor
Arterioscler. Thromb. Vasc. Biol., June 1, 2000; 20(6): 1512 - 1520.
[Abstract] [Full Text] [PDF]


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Circ. Res.Home page
A. Lymboussaki, B. Olofsson, U. Eriksson, and K. Alitalo
Vascular Endothelial Growth Factor (VEGF) and VEGF-C Show Overlapping Binding Sites in Embryonic Endothelia and Distinct Sites in Differentiated Adult Endothelia
Circ. Res., November 26, 1999; 85(11): 992 - 999.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
S. A. Stacker, K. Stenvers, C. Caesar, A. Vitali, T. Domagala, E. Nice, S. Roufail, R. J. Simpson, R. Moritz, T. Karpanen, et al.
Biosynthesis of Vascular Endothelial Growth Factor-D Involves Proteolytic Processing Which Generates Non-covalent Homodimers
J. Biol. Chem., November 5, 1999; 274(45): 32127 - 32136.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
S. Marchio, L. Primo, M. Pagano, G. Palestro, A. Albini, T. Veikkola, I. Cascone, K. Alitalo, and F. Bussolino
Vascular Endothelial Growth Factor-C Stimulates the Migration and Proliferation of Kaposi's Sarcoma Cells
J. Biol. Chem., September 24, 1999; 274(39): 27617 - 27622.
[Abstract] [Full Text] [PDF]


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Circ. Res.Home page
Y. Shizukuda, S. Tang, R. Yokota, and J. A. Ware
Vascular Endothelial Growth Factor–Induced Endothelial Cell Migration and Proliferation Depend on a Nitric Oxide–Mediated Decrease in Protein Kinase C{delta} Activity
Circ. Res., August 6, 1999; 85(3): 247 - 256.
[Abstract] [Full Text] [PDF]


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Cardiovasc ResHome page
J. R Kersten, P. S Pagel, W. M Chilian, and D. C Warltier
Multifactorial basis for coronary collateralization: a complex adaptive response to ischemia
Cardiovasc Res, July 1, 1999; 43(1): 44 - 57.
[Abstract] [Full Text] [PDF]


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Am. J. Pathol.Home page
R. Valtola, P. Salven, P. Heikkila, J. Taipale, H. Joensuu, M. Rehn, T. Pihlajaniemi, H. Weich, R. deWaal, and K. Alitalo
VEGFR-3 and Its Ligand VEGF-C Are Associated with Angiogenesis in Breast Cancer
Am. J. Pathol., May 1, 1999; 154(5): 1381 - 1390.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
M. E. Baldwin, B. Catimel, E. C. Nice, S. Roufail, N. E. Hall, K. L. Stenvers, M. J. Karkkainen, K. Alitalo, S. A. Stacker, and M. G. Achen
The Specificity of Receptor Binding by Vascular Endothelial Growth Factor-D Is Different in Mouse and Man
J. Biol. Chem., May 25, 2001; 276(22): 19166 - 19171.
[Abstract] [Full Text] [PDF]


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Physiol. GenomicsHome page
S. C. FRANCIS, M. K. RAIZADA, A. A. MANGI, L. G. MELO, V. J. DZAU, P. R. VALE, J. M. ISNER, D. W. LOSORDO, J. CHAO, M. J. KATOVICH, et al.
Genetic targeting for cardiovascular therapeutics: are we near the summit or just beginning the climb?
Physiol Genomics, December 21, 2001; 7(2): 79 - 94.
[Abstract] [Full Text] [PDF]




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