Characterising changes in the lipidome and metabolome of mammalian cells as a consequence of pharmacological induced stress
Advisory team: Dr Joanna Denbigh (80%), Professor Peter Gardner (10%)*, Dr Nick Lockyer (10%)*
Deadline: 31 March 2017
Characterising changes in the lipidome and metabolome of mammalian cells as a consequence of pharmacological induced stress affords new insight into biochemical processes associated with disease progression and the treatment thereof. Many diseases are linked with abnormal lipid metabolism. The chemical profiling of diseased and stressed biological systems through lipidomics and metabolomics is a powerful approach to understanding and ultimately controlling biological function.
The project will combine a number of analytical techniques to provide a multi-dimensional approach to the study of cellular metabolites and lipids. Cell lines will be cultured by the student in the BRC dedicated cell culture facility at The University of Salford and subsequently extracted in-house for HPLC-MS and GC-MS untargeted and targeted analysis of biomarkers  and prepared as intact fixed cells (methanol and/or formalin fixation) for imaging. Method development to ensure efficient extraction and fixing protocols will be explored .
Probing drug-cell interactions with spectroscopic techniques has become increasingly popular and can contribute to the understanding of the mode of action of the drug at a cellular level . The Fourier Transform Infrared (FTIR) imaging spectrometer in the Gardner group of The University of Manchester will allow biochemical changes within drug treated cells to be imaged at cellular resolution .
Time of flight secondary ion mass spectrometry (ToF-SIMS) is a powerful surface analysis technique which has seen much development in biological applications in recent years making ToF-SIMS a novel tool for probing the surface of biological materials . Recently there have been a number of reports into the localisation of metabolite and lipid species in various mammalian tissues using ToF-SIMS and other mass spectrometry imaging (MSI) techniques. The unique advantage of SIMS amongst MSI techniques is its sub-cellular spatial resolution, potentially allowing single cell metabolite analysis. ToF-SIMS will be carried out on dedicated instrumentation in the Lockyer group of The University of Manchester.
ToF-SIMS and LC-MS/GC-MS spectral data will be compared and contrasted to verify results obtained and to further explore the complementary possibilities of these techniques. Multivariate methods of data analysis will assist in data reduction and interpretation for the large and novel data sets that will be acquired. Image registration techniques will be employed to overlay data to provide added insight into localised biochemical pathways in response to stress and disease.
Further information is available here (project listed under Translational Medicine) www.salford.ac.uk/study/postgraduate/fees-and-funding/funded-phd-studentship/school-of-environment-and-life-sciences
The studentship is fully funded for UK/EU candidates. Successful candidates will receive a bursary of £14,553 tax free for up to three years and will also have tuition fees paid.
Candidates should have a first, or upper second class, honours degree and/or Masters degree and applications must be made before 31st March 2017.
Self-funded students with an interest in working in this field are welcome to contact Joanna Denbigh (J.L.Denbigh@salford.ac.uk) for further information.
* This project benefits from access to state of the art analytical instrumentation within The Manchester Institute of Biotechnology at The University of Manchester.
- Griffiths, W.J., et al., Targeted Metabolomics for Biomarker Discovery. Angewandte Chemie International Edition, 2010. 49(32): p. 5426-5445.
- Lyng, F., E. Gazi, and P. Gardner, Preparation of Tissues and Cells for Infrared and Raman Spectroscopy and Imaging. Biomedical Applications of Synchrotron Infrared Microspectroscopy, ed. D. Moss. 2011, Cambridge: RSC. 147-191.
- Derenne, A., M. Verdonck, and E. Goormaghtigh, The effect of anticancer drugs on seven cell lines monitored by FTIR spectroscopy. Analyst, 2012. 137(14): p. 3255-3264.
- Pilling, M. and P. Gardner, Fundamental developments in infrared spectroscopic imaging for biomedical applications. Chemical Society Reviews, 2016. 45(7): p. 1935-1957.
- Denbigh, J.L. and N.P. Lockyer, ToF-SIMS as a tool for profiling lipids in cancer and other diseases. Materials Science and Technology, 2015. 31(2): p. 137-147.