Metabolomics for Bioscience Research

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The significant global investment in genomic and proteomic tools for biochemical sciences has led to a rapid increase in our understanding of the genetic basis of cellular function and the influence of genetic changes on the proteome. However, downstream effects on metabolism remain significantly under-investigated but hugely important as they can manifest in changes to energy production, cell maintenance, proliferation and signalling. It is also in the metabolome that a direct interface with the external environment takes place. Metabolomics aims to provide comprehensive analysis of the entire small molecule component of biological samples from cells to whole organisms. Comprehensive metabolomics can be used as a discovery tool to assess the impact of upstream alterations to genetic and proteomic expression as well as the effects of environmental inputs on cellular metabolism. In Oxford Chemistry we have recently pioneered metabolomics of central energy metabolism which uses ion-exchange chromatography-mass spectrometry. Although very effective it can only be used for physiologically ionic and highly polar metabolites and does not give wide metabolome coverage. Furthermore it is funded for medical and clinical sciences and very heavily used for this. Researchers in the biosciences at Oxford desperately need a dedicated metabolomics platform to focus on fundamental biology, plant science and physiology. For example in the applicant's research in plant sciences metabolic profiling will benefit the investigation of plant-pathogen molecular relationships and normal functioning symbiotic relationships including nitrogen fixing bacteria interactions. Also studying the behaviour and functioning of plant metabolic networks to increase crop yield and resistance to disease. In physiology metabolomics will be used to study ketone metabolism, dietary performance response, hormone secretion by the gut and metabolism of infection. In fundamental biology and chemical biology metabolomics will play a key role in understanding mechanisms of sucrose signalling, algal responses to hypoxia, the interaction of redox balance and genetic studies interpreting the effects of DNA modifications on cellular pathway function and on artificial DNA synthesis to understand nucleic acid chemistry in vivo, develop therapeutic interventions and understand epigenetic regulation. We propose acquisition of a state-of-the-art Ion Mobility Mass Spectrometry system coupled to Ultra-Performance Liquid Chromatography (LC-IMS-MS/MS). This platform has excellent metabolite coverage of lower polarity metabolites including, importantly, lipids and plant secondary metabolites, using reversed phase chromatography. Ion-mobility capabilities will enhance metabolome coverage and increase confidence in compound identifications. The instrument will therefore provide an exceptional platform for metabolome-wide profiling. Furthermore this will complement other non-metabolomics capabilities dedicated to research in the biosciences enabling systems level analyses from genome expression to metabolome coverage. The proposed instrument does not overlap, but instead will be synergistic with existing capabilities enabling current and future BBSRC researchers from across the biosciences to move forward the frontiers of knowledge in systems biology, chemical biology, and the physiological and plant sciences. The new instrument will be integrated into the existing mass spectrometry laboratories (MS-SRF) in the Department of Chemistry in Oxford. It will be multi-user, enabling BBSRC-funded research groups in Oxford and the UK, to have dedicated access. The instrument will support research projects spanning a range of BBSRC strategic research priority areas, and will become an integrated piece of equipment in the Interdisciplinary Biosciences Doctoral Training Programme, making it available to researchers at other sites including Oxford Brookes University and the Diamond Light Source.