Cellular senescence is a mechanism that is activated in stressed cells to prevent the replication of damaged DNA. Senescent cells are found in vessel walls as well as in other tissues and have been shown to play a role in ageing and age-related diseases such as cardiovascular disease. Senescent vascular cells accumulate interleukin-1 as part of their secretory associated phenotype ( SASP) and may release this in the surrounding milieu causing damage to surrounding cells and in the local microenvironment. The release of interleukin-1 from vascular cells is a key regulated process and is understudied.
Fisetin is a flavonoid which has been proposed to extend health and lifespan. The mechanisms by which this might occur in vascular cells and in atherosclerosis are understudied.
The hypothesis for this project is that Fisetin prevents senescence in vascular endothelial cells via a mechanism involving the proinflammatory cytokine interleukin-1. In the project, the student will get a thorough training in vascular cell biology by culturing primary vascular cells, inducing senescence by different methods (doxorubicin, irradiation of hydrogen peroxide), assaying senescence ( beta galactosidase, P21, H2AX staining etc) and measuring IL-1 synthesis by qRTPCR and IL-1 protein release by ELISA. The effect of Fisetin will be studied on this process alongside the effects of the drug Anakinra or a gasdermin inhibitor to try to prevent IL-1 activity and release. In depth cell biology will be carried out to understand the mechanism of action of Fisetin in relation to involvement of the NLRP3 inflammasome and other cellular components.
To press forward with new hypotheses if Fisetin prevents senescence, the student will generate data from vascular cells +/- Fisetin using RNA seq, validating key findings by qPCR. The student will also study the pathology of already obtained arterial tissues from atherosclerotic mice +/- Fisetin to measure the number of senescent vascular cells in these groups. The expected value of the project is that Fisetin may be useful as a preventative or adjunct therapy in patients with coronary artery disease.
Entry Requirements:
Candidates must have a first or upper second class honors degree or significant research experience.
How to apply:
Please complete a University Postgraduate Research Application form available here: https://www.sheffield.ac.uk/postgraduate/phd/apply/applying
Please clearly state the prospective main supervisor in the respective box and select Infection, Immunity and Cardiovascular Science as the department.
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