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Volume 2
     

Therapeutic strategies using VEGF substitutes and gene therapies to maintain the integrity of the arterial wall (VEGF Strategies)

   
Project

QLK3-2002-01955

Cell factory area

3.1.3

EU Contribution

1,713,720 Euro

Duration

36 months

Type

RS

Starting date

01 December 2002

Keywords
arterioprotective therapies
artherosclerosis
endothelial cells
intimal hyperplasia
vasculoprotective functions
ABSTRACT

The partners have proposed a new hypothesis whereby vascular endothelial growth factor (VEGF) is arterioprotective in intimal hyperplasia and atherosclerosis. That cell signalling of the VEGF family, which underlines the production of NO and PGI2, is the basis of the arterioprotective effect, will be elucidated. Peptides will be synthesised and used as probes and as substitutes for the VEGF receptors in functional studies. The peptides will be the basis of future cytoprotective medicines. Rabbit and mice models of VEGF protective functions will be used. Novel VEGF- and PlGF-induced genes will be discovered leading to novel therapies.

Transgenic mouse models lacking or expressing mutant forms of the VEGF family of genes will be used to evaluate the vasculoprotective role in intimal hyperplasia and atherosclerosis. This project will elucidate the mechanisms involved in arterioprotection by VEGF and produce novel arterioprotective therapeutics.

OBJECTIVES
  • evaluate the effect of VEGF-A, -B, -D, -E, PlGF, and novel peptide VEGF substitutes on intimal thickening and restenosis in rabbit carotid arteries and aorta.
  • delineate the intracellular signalling pathways underlying VEGF-induced NO synthesis, PGI2 production and VEGF-dependent cell survival and leukocyte adhesion to endothelium.
  • define the receptors mediating the vascular protective functions of VEGF.
  • evaluate the contribution of other cellular mechanisms, including the novel possibility that VEGF may have direct effects on vascular smooth muscle cells.
  • study requirements for binding to and activation of the VEGF receptors.
  • characterisation of expression of VEGF-regulated genes as potential novel protective therapies.
  • expression of VEGF-regulated genes in vivo.
  • functional analysis of VEGF-regulated genes.
  • through novel chemistry, generate leads for a VEGF agonist and receptor activator.
DESCRIPTION OF THE WORK

The signalling pathways of VEGF-A, -B, -C, -D, -E and PlGF will be analysed along with the receptors involved. The role of PKC and FAK in VEGF-mediated NO and PGI2 production in endothelial cells will be studied using anti-sense oligonucleotides to specific RNAs of isoforms of PKC. Constitutively active and dominant negative PKC and FAK constructs will be produced by us for this purpose. Specific receptors mediating arterioprotection will be assessed using VEGF subtypes and cell lines expressing receptors for specific VEGF subtypes.

Using DNA screening and Northern blot analysis, VEGF stimulated gene expression in endothelial cells will be studied. The cellular mechanisms of action of selected upregulated genes will be delineated.

Novel chemistry will produce peptides which will identify receptor binding domains and requirement for receptor dimerisation to initiate the cell signalling cascade.

The agonist properties of these peptides will be tested in vitro. VEGF receptor tyrosine kinase (KDR) activators will be identified.

Transgenic mouse models will be used to assess the roles of the VEGF family and PlGF in atherosclerosis and intimal hyperplasia.

The arterioprotective properties of the novel genes identified and of peptides and KDR activators will be assessed in appropriate in vivo models. In particular, the effect of overexpression of the identified genes will be assessed in the "collar model" of intimal hyperplasia, including an assessment in vivo of whether the specific effect involves NO.

A full range of animal models will be used to assess and select genes and peptides.

DELIVERABLES
  • comprehensive understanding of cellular mechanisms mediating VEGF vasculoprotective functions in vitro and in in vivo.
  • understanding of the role of specific VEGF receptors in vasculoprotection.
  • we will develop new therapeutics that are effective in controlling disease in animal models of intimal hyperplasia and atherosclerosis.
CONSORTIUM
COORDINATOR
 
Prof. John Martin
Department of Medicine, BHF Laboratories
University College London
5 University Street,
London WC1E 6JJ, United Kingdom
Tel: +44-20-7679 7339
Fax: +44-20-7679 6379
john.martin@ucl.ac.uk
 
PARTNERS
 
Prof. Peter Carmeliet
Center for Transgene Technology and Gene Therapy
Flanders Interuniversity Institute for Biotechnology (VIB)
Campus Gasthuisberg O&N
Herestraat 49,
3000 Leuven, Belgium
Tel: +32-9-244 6611
Fax: +32-9-244 6610
Rudy.dekeyser@vib.be

Prof. Seppo Ylš-Herttuala
A.I. Virtanen Institute
University of Kuopio
Neulaniementie 2,
70211 Kuopio, Finland
Tel: +358-17-162 075
Fax: +358-17-163 751
Seppo.Ylaherttuala@uku.fi

Dr Johannes Waltenberger
Department of Internal Medicine II
Ulm University Medical Center
University of Ulm
Robert-Koch-Str.8,
89081 Ulm, Germany
Tel: +49-731-500 31158
Fax: +49-731-500 27019
johannes.waltenberger@medizin.uni-ulm.de

Dr David Selwood
Ark Therapeutics Ltd
1 Fitzroy Square
London W1T 6DE, United Kingdom
Tel: +44-20-7679 6716
Fax: +44-20-7679 6799
d.selwood@ucl.ac.uk