2024-W4: Demystifying Stable Isotope Labelling

Presenters: ▪ David de Souza, Metabolomics Australia ▪ Bart Ghesquiere, VIB ▪ James MacRae, The Francis Crick Institute Description: Metabolite profiling and targeted metabolomics approaches are now routinely applied by researchers wanting to understand the physiology of their cell, tissue, organism, or biofluid of choice. One of the largest challenges faced by metabolomics researchers in any study is in relating changes in identified metabolites to their roles in a given biological system. Understanding this relationship is critical for understanding the metabolic mechanisms in that biosystem. Stable isotope labelling metabolomics has rapidly emerged as a powerful tool in unravelling the precise structure and activity of metabolic pathways and how these influence/are caused by the physiological state of the biosystem. Such techniques can reveal subtle alterations in metabolic activity that would not be possible with unlabeled approaches. This extra depth of information can provide valuable insight into the mechanisms that underlie various physiological conditions and aberrant processes, including infection and disease. However, this technique is perhaps not used as widely as it might. This workshop seeks to show how metabolic labelling studies are readily approachable and easily interpretable when performed correctly. We will show you how to design, run, and analyze stable isotope labelling experiments. We will explain how these simple workflows are often sufficient for significant biological insight. This workshop is a mixture of traditional tutorials and some software demonstrations (laptops not essential, but would be useful). Workshop Objectives: ▪ Know how to design a labelling project, including the essentials to consider and potential pitfalls ▪ Know the basics of analysis of labelling data, including a run-through of some of the available LC-MS and GC-MS software ▪ Understand how to interpret data, including the importance of mass isotope distributions Learning Outcomes: ▪ Learn that stable isotope labelling projects are easy to design, perform, and interpret ▪ Labelling experiments can bring intricate detail to many metabolomics projects, whether untargeted or targeted, polar or lipid-focused ▪ Key principles behind data analysis workflows for labelling experiments, and how these differ when performed by LC-MS or GC-MS