Abstract
This is a perspective from the peer session on stable isotope labelling and fluxomics at the Australian & New Zealand Metabolomics Conference (ANZMET) held from 30 March to 1 April 2016 at La Trobe University, Melbourne, Australia. This report summarizes the key points raised in the peer session which focused on the advantages of using stable isotopes in modern metabolomics and the challenges in conducting flux analyses. The session highlighted the utility of stable isotope labelling in generating reference standards for metabolite identification, absolute quantification, and in the measurement of the dynamic activity of metabolic pathways. The advantages and disadvantages of different approaches of fluxomics analyses including flux balance analysis, metabolic flux analysis and kinetic flux profiling were also discussed along with the use of stable isotope labelling in in vivo dynamic metabolomics. A number of crucial technical considerations for designing experiments and analyzing data with stable isotope labelling were discussed which included replication, instrumentation, methods of labelling, tracer dilution and data analysis. This report reflects the current viewpoint on the use of stable isotope labelling in metabolomics experiments, identifying it as a great tool with the potential to improve biological interpretation of metabolomics data in a number of ways.
Highlights
The major interest in stable isotope labelling for metabolomics stems from its ability to aid in the measurement of dynamic activity of metabolic pathways, in order to provide mechanistic explanations for the perturbations in steady-state metabolite levels observed in classical metabolomics studies
Whether being used as a hypothesis-generating holistic approach, or for hypothesis-driven analysis of specific pathways, stable isotope labelling is a powerful tool that can enhance the biological interpretation of metabolomics data
Stable isotope labelling can help alleviate a number of issues associated with metabolomics data analysis and provide confidence in assigning correct identification to metabolite features
Summary
Metabolomics has emerged as a powerful approach for enabling the study of the intricate biochemistry of cells, organisms, and systems in response to different conditions such as stress, disease, or nutrition This detailed analysis of metabolite profiles can provide functionally relevant biochemical information about biological processes. Combining the use of stable isotope labelled metabolic precursors (tracers), and modern analytical techniques (such as high resolution chromatography and mass spectrometry) for separation and detection of chemical species permits characterization of the metabolism of a broad range of biological systems This can include direct and accurate measurement of turnover of a specific metabolite or even determining entire pathway fluxes in vitro or in vivo [4,5,7]. Robust untargeted metabolome characterization, leading to pathway mapping throughout the metabolic network has been made possible by combining advanced stable isotope labelling methods with improved genome annotations [14,15,16]
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