swdtp - University of Leeds

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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/).
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