Ultrahigh sensitivity metabolomics: opening a window into the metabolic evolution of a sentinel invertebrate through the industrial revolution

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METABOLOMICS is a proven technology for discovering mechanisms of how organisms respond to stress. However, the mass spectrometry methods employed do not yet provide sufficient analytical sensitivity to measure metabolites in the very small samples that are often encountered in environmental studies. Nanoscale liquid chromatography-mass spectrometry (nanoLC-MS) is applied extensively in proteomics - offering high sensitivity - but is not yet used in metabolomics due tomany technical challenges. Here we propose to partner with Thermo Fisher Scientific to establish and optimise novel nanoLC-MS methods to enable transformative investigations in environmental toxicology. Daphnia are freshwater invertebrates that play a central role in foodwebs and are widely studied in ecotoxicology. They can form dormant eggs that accumulate in sediments - which can hatch into functional animals many centuries later - allowing reconstruction of responses to past environmental change. This is termed resurrection ecology, and the current record isthe resurrection of 700 yr old Daphnia. The biochemical study of dormant eggs in sediment cores therefore offers a unique opportunity to investigate pollutant impacts on animal fitness from well before and throughout the industrial revolution, up to modern timescales of intensive agriculture and high pesticide usage. Yet Daphnia eggs weigh <1 mg, undetectable by current metabolomics technologies. Building on our decade-long track record in metabolomics, we propose to establish and optimise nanoLC-MS metabolomics technologies that are sensitive, reproducible and robust, and then apply these to discover the biochemical basis of Daphnia fitness through historically changing pollutant levels. Hence we will discover associations betweenenvironmental pollution and the health of this sentinel freshwater species.Our CASE PARTNER, Thermo Fisher Scientific (TFS), is a world leader in the development of nanoLC-MS and has identified metabolomics as a priority area. In 2013, TFS formed a Technology Alliance Partnership (TAP) with the University of Birmingham, the first such partnership within Europe. This proposal builds upon five existing iCASE awards with TFS as part of their commitment to graduate training. Collectively this team will provide SPECIALIST TRAINING: in nanoLC-MS, including unparalleled access to current and pre-released metabolomics technologies (at TFS); in metabolomics, environmental toxicology and analytical sciences (Viant lab; the largest group in environmental metabolomics nationally).TRANSFERABLE SKILLS will be taught at both Birmingham, through the extensive courses in the Biosciences Graduate Research School, and at TFS, including business awareness, project management and financial training. This training will be truly MULTIDISCIPLINARY to enrich the student experience. Furthermore the main supervisor is highly experienced,having completed 5 PhDs in the past five years and with 5 current students, several of which are/were NERC iCASE.