Liquid Chromatography-Mass Spectrometry for the Analysis of Biological Small Molecules in Metabolomic, Nutritional, and Toxicological Studies.

This proposal requests support for a piece of equipment called a liquid chromatography - mass spectrometer (LC-MS). This equipment allows researchers to measure the mass of particles, and to find the basic chemicals that make up a biological sample such as blood, tissue or food. The LC-MS requested in this proposal will support research from multiple scientific disciplines, and on several distinct projects of great importance to the health of the population:

Example Project 1: Given that 340 million people worldwide suffer from type 2 diabetes, finding new treatments is essential. White fat tissue stores energy as fat and releases signals into the blood which communicate with other organs, regulating whole-body metabolism. However, not all fat cells are identical. A second fat cell population, brown fat cells, regulate body temperature by "burning" stored fats, producing heat. A third fat cell population, beige fat, can switch from behaving like white fat cells (storing fat) to functioning like brown fat cells (burning fat), a process called "browning". In humans, beige fat may form the majority of this heat-producing tissue, and is also found within white fat tissue. Beige fat has anti-diabetic properties which may, in part, occur through release of signals into the blood which alter metabolism in tissues such as liver, muscle and white fat.
This research will use the requested LC-MS to determine whether browning of white fat tissue results in metabolite signal secretion, and to identify these signals. The capacity of metabolite signals to induce browning in neighbouring white fat tissue and to signal, via the blood, to other organs to regulate whole-body metabolism, will be investigated. This research may identify beige cell-specific metabolite signals, with anti-obesity properties, and provide new anti-diabetic therapies.

Example Project 2: Food constituents can have profound effects on wide ranging aspects of health including gut function, control of blood glucose and our ability to think. Wheat and other food crops contain naturally occurring compounds, called phenols, which may have anti-oxidant effects that reduce cellular damage to DNA known to be important in cancer and ageing. They may also improve gut function. This project will use the LC-MS to measure the antioxidant phenols, and micronutrients in models of gut function to understand how readily these antioxidant phenols are absorbed from food, determine ways of improving their uptake, and to measure the absorption of other micronutrients across the gut in the presence of the phenols to determine how they affect gut nutrient uptake.

Example Project 3: Food contaminants and toxins are a major risk to human health and have particular impact on vulnerable communities in low income countries. A particular group of toxins know as mycotoxins, derived from fungus, can contaminate food crops such as maize and wheat and have been linked to reduced child growth and development, and the development of cancer. This project will use the requested LC-MS to measure levels of the toxins and markers of exposure to the toxins in human populations from Africa to understand distribution patterns of contamination in different crops, storage conditions and geographic locations and to assess the effectiveness of interventions to reduce mycotoxin risk in Africa. Therefore contributing to the safety and security of the global food industry.

Example Project 4: Exposure to environmental pollution in the form of particulates from traffic in the air can alter the way in which blood clots and contribute to deep vein thrombosis. This project will use the requested LC-MS to measure markers of exposure to the particle pollutants in blood samples of the population to understand the effects of pollution on cardiovascular health.

This proposal seeks a Waters Xevo TQ-XS Liquid Chromatography-Mass Spectrometry (LC-MS) system to enhance analytical chemistry applied specifically to biological small molecules across research priorities central to both the Faculty of Mathematics and Physical Sciences (MAPS) and the Faculty of Medicine and Health (FMH) at the University of Leeds.
The LC-MS is a liquid chromatograph coupled to a tandem quadrupole mass spectrometer capable of both electrospray and atmospheric pressure chemical ionization, utilized for the mass separation, identification and quantification of a range of small molecules, including metabolites, lipids, drugs and chemical contaminants within biological samples. This LC-MS system offers advanced sensitivity and reliability of detection with straightforward maintenance and intuitive data processing. The LC-MS is capable of cross-discipline small molecule applications including nutritional, cellular, toxicological and drug metabolite analysis. Within the FMH the LC-MS system will support research including metabolomics approaches in tissue culture and rodent models, and human intervention studies, to investigate the pathophysiology of cardiometabolic diseases; the metabolism of adipose tissue; and the contribution of nutritional and environmental factors to the prevention of metabolic disease. Within MAPS the LC-MS will support research in food safety, from chemical toxin analysis in food matrices, to biomonitoring of human exposure to dietary chemical contaminants; a vital step towards assessing and preventing unsafe exposure from food and environment, and understanding how exposure contributes to non-communicable disease in society.
The LC-MS system will support research of global importance to population health and wealth creation whilst aligning with BBSRC Strategic Research Priority, "Bioscience Underpinning Health" and BBSRC Responsive Mode Priorities; "Food, Nutrition and Health", and "Healthy Ageing across the Lifecourse".

This statement sets out how the Liquid Chromatography - Mass Spectrometer (LC-MS) requested in this proposal will be translated into improvements in human health, and benefits to UK competitiveness and wealth. Objectives include the dissemination of the research to potential beneficiaries, policy makers, industry, and the public through both direct and indirect engagement.
Research generated using the LC-MS will create scientific impact for a scientific audience. The outputs will include peer-reviewed publications throughout the instrument lifetime, adhering to BBSRC open access policy. The applicants have experience publishing in high impact journals (e.g. Cell Metabolism, Diabetes and The Lancet). Impact will also include conference proceedings. This will be complemented by PI and Co-I membership of scientific bodies (e.g. European Association for the Study of Diabetes, EASD). The research will be reported at both broad interest international conferences (e.g. EASD Annual Meeting) and specialised conferences (Metabolomics Society). Impact will also be via data sharing. Metabolomic data will be made accessible on databases (e.g. MetaboLights) not more than 2 years post study completion. These outputs will impact wider research into the role of adipose tissue in the aetiology and treatment of metabolic diseases of global importance; the understanding of bioactive nutritional small molecules and their role in human health and disease; and the identification of food contaminants and toxins and their importance to food safety and toxicology.
The research supported by the LC-MS may lead to the discovery of novel therapeutics for metabolic diseases, yield improvements in food processing and selection with impacts on population health and food industry wealth, and may have significant impact on food safety and toxicology, and the understanding of the impact of pollution on cardiovascular health. Intellectual property (IP) generated from these research approaches will be exploited through patent protection, commercial partnering and licensing. Interaction with industry may facilitate the transition from discovery to commercialisation; this transition can run to decades. However, shorter term goals such as patent application will occur over a shorter period (3 - 5 years). The generation, protection and exploitation of IP will be facilitated by interactions with the university IP agent.
Research using the LC-MS may influence national and international public policy. This research may influence dietary and dietary supplement advice and influence policy on food system and pollution management and food safety. This will have a significant impact on the health of society and could reduce the economic burden on healthcare by informing policy makers in Public Health England, UK Departments of Health, Department for Environment Food and Rural Affairs, NICE and the NHS.
The LC-MS would have a continuing impact on the UK economy by providing a training platform to instruct graduate students in the application of analytical chemistry techniques in biological research. Projects will be offered through PhD training schemes (MRC and BBSRC Doctoral Training Partnerships) offering training for the next generation of scientists in cutting edge mass spectrometry-based biological techniques.
Infrastructure Impact - In the long-term (5 - 10 years) the LC-MS will provide the catalyst for impact to university infrastructure in the form of a small mass spectrometry and metabolic phenotyping facility/core, closely linked to the School of Food Science and Nutrition (FSN) strategic plan to build a "food safety" suite. The equipment will contribute to shaping new research direction in the FSN towards an international leading research centre in food safety, dietary health, and sustainable food systems.
Regional Impact - The LC-MS will also impact the research of our N8 regional partners in the most research intensive universities in Northern England.