Redox-dependent regulation of endothelial cell physiology and angiogenesis during ischemia Supervisory team: Dr Giordano Pula (Department of Pharmacy and Pharmacology, University of Bath); Prof Costanza Emanueli (School of Clinical Sciences, University of Bristol); Prof Stephen Ward (Department of Pharmacy and Pharmacology, University of Bath); Prof Harry Mellor (School of Biochemistry, University of Bristol). Angiogenesis is one of the most important physiological processes in the human body and is a critical step for the regeneration of ischemic tissues, such as infarcted myocardium or diabetic limbs. Despite its importance, many aspects of the regulation of angiogenesis are still unknown. For example, reduced blood supply in ischemic tissues is known to stimulate angiogenesis in a reactive oxygen species (ROS)-dependent manner. Nonetheless, the following important questions on ischemia-dependent regulation of angiogenesis remain unanswered: 1) Which angiogenic factors and which redox-regulating proteins (i.e. ROSgenerating enzymes, ROS-degrading enzymes, ROS scavenging proteins) are upregulated during ischemia and might participate in the stimulation of reparative angiogenesis? 2) What is the role of redox-regulating proteins in the regulation of ROS generation and cell functions in endothelial cells during ischemia? 3) Can we mimic the redox state of endothelial cells during ischemia and induce a pro-angiogenic phenotype in cultured cells? In the first phase of the project, we will test hind limb tissue by qPCR for the expression of angiogenic factors and different redox-regulating proteins at different stages of ischemia (i.e. 0, 1, 2, 4, 7 and 14 days after artery ligation). We will then generate a mathematical model of the temporal progression of ischemia in terms of angiogenic factor and redox-regulating protein expression. The redox conditions observed in the ischemic muscle will then be mimicked in human endothelial cells in vitro by recombinant overexpression or siRNAdependent down-regulation of redox-regulating proteins. The effects on ROS generation, cell proliferation, cell apoptosis, angiogenic markers expression and capillary tube formation will then be tested. The effects on cytoskeletal organization and cell motility will also be tested. The student will utilize the expertise in endothelial cell signalling and angiogenesis analysis expertise of the laboratory of Dr Pula, the in vivo and vascular surgery skills of the laboratory of the second supervisor Prof Emanueli, and the signal transduction and cell imaging capability of the labs of the co-supervisors (Prof Ward and Prof Mellor, respectively). Figure 1: Two important techniques utilised in this studentship: in vitro capillary tube formation by human umbilical vein endothelial cells (HUVECs) (left) and surgical induction of hind limb ischemia in mice (right). For any enquiry please contact Dr Giordano Pula: [email protected] Deadline for online applications is Friday 10 January 2014 (http://www.bris.ac.uk/swdtp/apply/).