Realising the routine measurement of normal human physiology: A Case Study in Probing the function of the human gut microbiota with stable isotopes
Lead Research Organisation:
Scottish Universities Environmental Research Centre
Department Name: SUERC
Abstract
We lack tools which give deep insight into normal human physiology in vivo. We are reliant on cell and animal models which do not capture human biological variability - a fundamentally important aspect of biology which if understood would lead to more targeted approaches to health.
Stable isotopes (SI) are non-radioactive minor versions of elements in biology which are measured by mass spectrometry (MS) and used to trace the flow of molecules though complex systems. Hitherto, SI studies have largely traced single molecules whereas to fully capture the complex dynamics of real life, we need to look at whole systems. In previous BBSRC funded work, we have used low-level SI labelled food to understand diet-gut-microbiome interactions. By targeting the SI labelled (13C) metabolites available through current technology, we demonstrated that a genetic mutation in the starch branching enzymes of peas reduced glucose from digestion and increased carbohydrate fermentation by the gut microbiota in humans (doi.org/10.1038/s43016-020-00159-8). We have a unique biorepository of samples (plasma, urine and stool) from this study which fed complex food structures (cell wall components, starch, protein) but we have lacked the isotope technology to fully probe these samples for all SI features.
The new ThemoFisher-Scientific (TFS) Orbitrap Exploris Isotope Solutions (OEIS) MS is a game-changing solution to quantifying unique low-level 13C labelled features with unparalleled sensitivity providing in-depth information about gut microbiota function and linking directly dietary substrates, gut function and microbiome function in vivo. We will use our unique 13C pea study samples to establish proof of concept of this new approach. This project goes significantly beyond the current state of the art to establish deep SI-labelled metabolite profiling in normal "free living" human studies with potential to trace many metabolic pathways simultaneously.
Stable isotopes (SI) are non-radioactive minor versions of elements in biology which are measured by mass spectrometry (MS) and used to trace the flow of molecules though complex systems. Hitherto, SI studies have largely traced single molecules whereas to fully capture the complex dynamics of real life, we need to look at whole systems. In previous BBSRC funded work, we have used low-level SI labelled food to understand diet-gut-microbiome interactions. By targeting the SI labelled (13C) metabolites available through current technology, we demonstrated that a genetic mutation in the starch branching enzymes of peas reduced glucose from digestion and increased carbohydrate fermentation by the gut microbiota in humans (doi.org/10.1038/s43016-020-00159-8). We have a unique biorepository of samples (plasma, urine and stool) from this study which fed complex food structures (cell wall components, starch, protein) but we have lacked the isotope technology to fully probe these samples for all SI features.
The new ThemoFisher-Scientific (TFS) Orbitrap Exploris Isotope Solutions (OEIS) MS is a game-changing solution to quantifying unique low-level 13C labelled features with unparalleled sensitivity providing in-depth information about gut microbiota function and linking directly dietary substrates, gut function and microbiome function in vivo. We will use our unique 13C pea study samples to establish proof of concept of this new approach. This project goes significantly beyond the current state of the art to establish deep SI-labelled metabolite profiling in normal "free living" human studies with potential to trace many metabolic pathways simultaneously.
