Objectives

The main objective of this Workpackage is the determination of the role of inflammation / thrombosis in macro- and microvascular CVD.

Task 1:  Investigation of A. carotis
Task 2:  Investigation of A. saphena
Task 3:  Investigation of stress responses in irradiated hearts
Task 4:  Investigation of inflammatory and thrombotic responses and
            changes of immune function after heart irradiation

Workpackage description

• In a first step, the changes in inflammatory and thrombotic gene expression in a.carotis will be quantified by using qRT-PCR and immunohistochemistry at different times after irradiation.
• Additionally, the changes in expression of the same inflammatory and thrombotic genes in the irradiated a.saphena by immunohistochemistry will be determined at different times after local irradiation.
• The expression of stress responses (hsp70, pro-inflammatory cytokines) in irradiated hearts will be investigated, some of which will be subjected, before sacrifice, to acute hypoxic and reoxygenation stress.
• Additionally, the stress response (HSP and HIF) in endothelial sheets after exposure to hypoxic, thermal or other shall be studied.
• The activation of the Rho/ROCK cascade in non irradiated vs irradiated hearts, which might constitute the main intracellular signalling pathway between oxidative stress and the downstream thrombogenic, inflammatory and fibrogenic response in cardiovascular disease will be investigated.
• The changes in inflammatory proteins in irradiated hearts at early and late times using immunohistochemistry and Western blotting as well as inflammatory gene expression by qRT-PCR will be studied.
• Finally, the following markers of an inflammatory or a thrombotic response will be analysed: selectin family, E/P-selectin ELAM-1, CD62; integrin family, VLA-1 -6; immunoglobulin supergene family,  PECAM-1, ICAM-1 VCAM-1, and class I heavy chain proteins; complementary adhesion molecules CD34, CD44 as well as Thy1 and MHCII, MCP-1, CD40, TM, KLF-2, PAI1 and vWF.
• Thrombocytes in heart microvessels will be detected with anti platelet glycoprotein IIIa antibodies in frozen heart sections.
• Levels of circulating ANP and EC apoptotic microparticles will be assessed in the serum at different times after local heart irradiation.
  

WP leader

Dr. med. G. Hildebrandt (Technical University of Leipzig) is deputy head of the Department of Radiation Oncology and head of the Radiobiological Laboratory. He has long-term experience in investigations of low dose radiation effects on inflammation (chronic, acute) in different species (human, rabbit, rat, mice) in vitro and in vivo.
The radiobiology research group in the department of radiotherapy has extensive experience in the study of biological effects of low dose irradiation, in the radiobiology of endothelial cells, of dendritic and T-cells and the biology of primary skin cells providing all necessary skills and techniques to perform the intended experiments. The radiobiology group is a partner in the NOTE integrated project and in projects funded by the German research foundation (DFG).

Participating consortium partners

Technische Universtität München, Germany
Netherland Cancer Institute, The Netherlands
Institut de Radioprotection et de Sureté nucléaire Villejuif, France
Maria Sklodowska - Curie Cancer Institute, Poland
Cardiovascular Research Institute, Maastricht, The Netherlands