Understanding the inflammatory response of endothelial cells to oxidised low density lipoproteins (360G-Wellcome-102290_Z_13_Z)

Atherosclerosis is a major cause of mortality and morbidity worldwide; it is an inflammatory condition in which lesions are packed with immune cells. One of the key steps in immune cell migration into the vessel wall is activation of overlying endothelial cells. Circulating oxidised low-density lipoprotein (LDL) is a major risk factor for atherosclerosis and a potent activator of endothelial cells. The intracellular pathways by which this pro-inflammatory activation is mediated are poorly understood. The aim of this study is to define this response of primary human arterial endothelial cells to LDL at the chromatin level. This will allow identification of transcriptional regulatory pathways that are triggered when endothelial cells are activated by oxidised-LDL. In the longer term, it is hoped this will allow identification of potential therapeutic targets in the management of atherosclerosis. The approach used will be based on the genome-wide isolation and sequencing of regions of open chromatin in endothelial cells using FAIRE (formaldehyde-assisted isolation of regulatory elements)-seq. The key goals of the project include: 1. Generation of FAIRE-seq data on ‘resting’ and activated endothelial cells 2. Generation of microarray data to identify genes that are up-/down-regulated in oxidised-LDLactivated endothelial cells 3. Bio-informatic analysis of this data to identify activated endothelial cell-specific regulatory DNA elements 4. Functional experiments to explore and validate these identified chromatin elements, including chromatin conformation capture as appropriate. The project will give me training in a range of techniques in high throughput sequencing, cellular and molecular biology and bioinformatics.