The most common form of localised amyloid occurs in the aorta (aortic medial amyloid; AMA) and is estimated to occur in 97% of Caucasian people over 50. The main constituent of AMA has been identified to be fragments thought to be derived from the proteolysis of milk fat globule-EGF factor 8 protein (MFGE8). The full pathological impact of AMA is unknown, but it is believed that extracellular amyloid accumulation contributes to age-related diminished elasticity of the aorta and may underlie the pathogenesis of sporadic thoracic aortic aneurysm, in turn leading to rupture.
The aim of this project is to identify which MFGE8 fragments form amyloid deposits in human tissue obtained during aneurysm and dissection surgery at Liverpool Heart and Chest Hospital using mass spectrometry (MS)-based methods. Following this conceivable mechanisms of cleavage of MFGE8 that could result in fragments identified in human tissue will be probed with a view to manipulating cellular MFGE8 proteolysis and thereby reduce disease-causing AMA.
Amyloid will be isolated from human tissue using two approaches optimised in the host lab; extraction and laser microdissection. MS techniques, including top-down analysis of intact proteins and ion-mobility-MS, will be used to define the sequence of those protein fragments present within the amyloid deposits and explore aggregation mechanism of the MFGE8 fragments. Following characterisation of MFGE8 cleavage products, bioinformatic and biochemical strategies will be used to evaluate the potential of different proteases, to generate the active amyloid forming protein variants. Training will be provided in all biochemical, biophysical and MS techniques required for the project.
The student will also be integrated within the Liverpool Aorta Biomechanics and Biochemistry Research Group (http://labb-group.com/) and Centre for Proteome Research (https://www.liverpool.ac.uk/pfg/).
Benefits of being in the DiMeN DTP:
This project is part of the Discovery Medicine North Doctoral Training Partnership (DiMeN DTP), a diverse community of PhD students across the North of England researching the major health problems facing the world today. Our partner institutions (Universities of Leeds, Liverpool, Newcastle and Sheffield) are internationally recognised as centres of research excellence and can offer you access to state-of the-art facilities to deliver high impact research.
We are very proud of our student-centred ethos and committed to supporting you throughout your PhD. As part of the DTP, we offer bespoke training in key skills sought after in early career researchers, as well as opportunities to broaden your career horizons in a range of non-academic sectors.
Being funded by the MRC means you can access additional funding for research placements, international training opportunities or internships in science policy, science communication and beyond. See how our current DiMeN students have benefited from this funding here: http://www.dimen.org.uk/overview/student-profiles/flexible-supplement-awards
Further information on the programme can be found on our website:
Studentships are fully funded by the Medical Research Council (MRC) for 3.5yrs
Stipend at national UKRI standard rate
Research training and support grant (RTSG)
Studentships commence: 1st October 2019.
To qualify, you must be a UK or EU citizen who has been resident in the UK/EU for 3 years prior to commencement. Applicants must have obtained, or be about to obtain, at least a 2.1 honours degree (or equivalent) in a relevant subject. All applications are scored blindly based on merit. Please read additional guidance here: https://goo.gl/8YfJf8
R.Q. Migrino, S. Truran, N. Karamanova, H. Davies, D.A. Franco, G. Serrano, T. Beach, J. Madine, Amyloidogenic medin induces endothelial dysfunction and vascular inflammation through the receptor for advanced glycation endproducts, Cardiovascular Research 2017, 113, 1389–1402
H. Amijee, C. Bate, A. Williams, J. Virdee, R. Jeggo, D. Spanswick, D. IC Scopes, J.M. Treherne, S. Mazzitelli, R. Chawner, C.E. Eyers, A. J. Doig The N-methylated peptide SEN304 powerfully inhibits Aβ (1–42) toxicity by perturbing oligomer formation (2012) Biochemistry, 51, 8338–8352