Tracking metabolic reprogramming through stable isotope-resolved metabolomics

Evidence suggests that the way cells consume and process nutrients, collectively known as metabolism, differs between healthy and diseased cells and can even change the way they communicate with adjacent cells, survive and grow. Changes in metabolism are very important in protecting us against dangers such as infections or cancer, but have also been implicated in neurodegenerative diseases and metabolic disorders such as obesity, type 2 diabetes, and metabolic syndrome. Both systemic and local metabolic alterations can also affect drug response and the acquisition of antibiotic or anti-cancer drug resistance.

This project will draw on the strengths of acquiring and setting up a specialised equipment known as mass spectrometer capable of determining the mass of analytes based on the motion of ions in electric and magnetic fields. Through setting up assays to measure different classes of core metabolites (e.g. amino acids, carbohydrates, nucleotides, lipids etc.) the first objective of this proposal is to provide the capability to track the rewiring of metabolism in the context of infection, immunity, metabolic disorders and cancer, and their organism-wide implications across different organs and tissues. This will assist with gaining insight into the metabolic interplay between host immune cells and pathogens or cancer cells and how these interactions ultimately affect disease outcomes. The second objective of this proposal is, to exploit metabolic profiles to identify more effective predictors of response to therapy and unveil novel metabolic dependencies and vulnerabilities.

This approach will not only provide novel mechanistic insights of disease pathogenesis, but could also enable biomarker discovery of metabolites that predict pathogenic processes and the response to targeted or untargeted therapies. In the longer term this knowledge will help identify weaknesses in the pathogen or the tumour's armour and ultimately unveil more effective therapeutic strategies where diet and metabolic targets could empower our body to better defend attacks by "foreign" invaders.

Metabolomics is a rapidly evolving analytical approach in life and health sciences. Stable isotope labelling, as part of metabolomics approaches, provides a unique picture of intracellular metabolism. Although untargeted metabolomics provides the abundance of different metabolites within metabolic pathways, several important metabolic changes do not result in an increase or a decrease in the metabolite level. Stable isotope tracing or fluxomics provides information not revealed by conventional untargeted metabolomics by measuring the rates of metabolite interconversion as a readout of metabolic enzyme regulation. This makes stable isotope tracer studies a powerful option to probe metabolic changes in complex biological systems. We are seeking funds to purchase a 6546 Liquid Chromatography Mass Spectrometer (LC-MS)-Quad Time Of Flight (QTOF) instrument from Agilent Technologies. This mass spectrometer will enable us to assay both carbon and lipid flux metabolism and is particularly designed for fluxomics thanks to the high resolution and high accuracy as well as supporting software. It provides quantitative information on metabolites with an accuracy and precision that no other existing system can match.