Plaque rupture is are 2 within EVGN network and consists of 5 workpackages (WP).

The specific objectives of the workpackages WP7 and WP12 that use the more advanced genomics/proteomics technologies are:

1. To perform analyses of datasets of partners 5, 13, 14 using the bioinformatics platform- developed during the past 2 years, including Genomatix Bibliosphere, gene set enrichment analysis (GSEA) and Compendium Module analysis, and implemented by Dr Oscar Volger, the EVGN-sponsored bioinformatician at AMC, Amsterdam.

2. To validate a number of candidates from the 750 validated microarray expression profiles for endothelial cells, smooth muscle cells, monocytes/macrophages/foam cells and T cells from different origins in vivo (healthy and lesion vascular versus inflamed non vascular and normal tissue) and ex vivo (culture models) source.

3. To establish DGE profiles of endothelial cells exposed smoke extracts, with or without prior stimulation with chronic shear, and DGE profiles of conductance arteries in vivo exposed to normal of no flow.

4. To evaluate the role of plasminogen system in high shear stress-induced vascular remodelling, using mice deficient in uPA or tPA or using uPAR/uPA/PAI contructs.

Workpackages WP8 and WP9 are oriented towards the development of mouse models of atherosclerosis that are required to bring forward novel strategies aimed at reducing plaque rupture, and to validate new genes identified as potentially involved in plaque stabilisation/destabilisation. The specific objectives of these workpackages are:

5. To study the kinetics of smooth muscle cell and macrophage migration and proliferation in the mouse brachiocephalic artery, to evaluate whether induction of smooth muscle apoptosis promotes plaque rupture in brachiocephalic arteries.

6. To start the generation of mice that express a tamoxifen-dependent Cre recombinase, Cre-ERT2, in monocyte/macrophage or CD4 T cell lineages due to targeted insertion of the Cre cDNA into their endogenous M lysozyme or CD4 locus, respectively. Preferably, mice on a C57BL6 background will be generated.

7. To start the generation of of Cathepsin-S floxed mice, Nedd4 floxed mice, Vasculin-knockout mice, and transgenic mice expressing the estrogen receptor  driven by the endothelial-specific promoter Tie2.

The workpackage WP10 is aimed at determining the role of specific subpopulations of T cells, B cells and dendritic cells (DC) in the protective immune response elicited during the initiation and progression of atherosclerosis using validated mouse models of human disease. Its specific objectives are:

8. To develop and validate experimental protocols for the in vitro generation of B cells, Treg and tolerogenic DC specific for plaque-related antigens (Heat Shock Proteins, apoptotic cell membranes, oxidized LDL and others).

9. To use newly developed compound knockout strains to assess the role of specific immune regulatory genes in atherosclerosis.

10. To develop and validate protocols of vaccination against atheroscleroisis in apoE-/- and LDLr-/- mice, based on the stimulation of plaque antigen-specific regulatory T cells.

The workpackage WP11 is a clinically-oriented workpackage that aims to identify patients at high risk for the development of atherosclerosis and its severe manifestions (gangrene, stroke, myocardial infarction, heart failure and sudden death) by using a combination of serum markers. Its specific objectives are:

11. To determine the predictive value of PlGF and sFlt prospectively using samples of the OPTIMAAL study.

12. To evaluate the predictive value of plasma levels of sPLA2 activity and lactadherin in large populations of subjects with no clinical symptomes and. An assay for lactadherin will have to first be build up for the clinical use.

13. To assess whether the ratio of apoptosis/cell death and repair (circulating endothelial progenitor cells) is informative for the prognosis of the patients with acute coronary syndromes.