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Table of contents
I Introduction/Scientific background
I.1. Atherosclerosis
I.1.a Physiopathology of atherosclerosis
I.1.b cell types involved in the plaque progression
I.2 High-Throughput technologies and atherosclerosis
I.3 Smoking and atherosclerosis
I.3.a The pathophysiology linking smoking and atherosclerosis
I.3.b Molecular pathways linking smoking to atherosclerosis
I.3.c Modeling cigarette smoke exposure in vitro
I.4 SASH1
I.4.a SASH1 gene
I.4.b SASH1 protein
I.4.c SASH1 in the literature, the quest for SASH1 function
I.5 Thesis project: from Genome Wide Analysis in the Gutenberg Health Study to in vitro experiments
I.6 Aim of the thesis and experimental strategy
II Article: SASH1 a new link between cigarette smoke and atherosclerosis
II.3 SASH1 ongoing experiments and unpublished results
II.3.a Materials and methods
II.3.b Detecting the SASH1 protein in HAECs
II.3.c SASH1 expression in vascular cells
II.3.d Testing the effect SASH1 knockdown on the SASH1 network
II.3.e Effect of HAECs stimulation with inflammation related compounds on SASH expression and its sub cellular localization
II.3.f Transcriptomic analysis of cigarette smoke condensate stimulated HAECs
II.3.g In silico study of SASH1 methylation
II.4 Searching for SASH1 partner proteins
II.4.a Yeast two hybrid (Y2H)
II.4.b Co-immunoprecipitation
II.4.c Mass spectrometry
III Discussion
References
ANNEXES
Annex 1: SLC39A8 (ZIP8) and cigarette smoking, genesis of a new project
I Introduction
I.1 SLC39A8 gene organisation
I.2 SLC39A8 protein (ZIP8)
I.3 Zip8 in the literature
I.4 Material and methods
II Results
II.1 ZIP8 expression in vascular cells and in the vascular wall
II.2 ZIP8 expression and the plaque formation
II.3 ZIP8 expression in human carotid plaque according to the smoking status
II.4 Investigating the interdependence of ZIP8 and Gas6 expression in human macrophages
II.5 Methallothionein and ZIP8 expression
III Discussion
Références
Annex 2. Highlights for the ZIP8 project
Annex 3. Hightlights of the SASH1 project
Annex 4. Verdugo et al 2013
Annex 5. Touat et al (publication in preparation)
