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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,
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
-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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T. A. PARTANEN, J. AROLA, A. SAARISTO, L. JUSSILA, A. ORA, M. MIETTINEN, S. A. STACKER, M. G. ACHEN, and K. ALITALO VEGF-C and VEGF-D expression in neuroendocrine cells and their receptor, VEGFR-3, in fenestrated blood vessels in human tissues FASEB J, October 1, 2000; 14(13): 2087 - 2096. [Abstract] [Full Text]
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