Projects

Context: 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.

Aims and Objectives: This project will combine low-level 13C labelling and OEIS workflows to explore the 13C metabolite landscape produced from the breakdown of food in the human gut. The objectives of this project are to: 1) establish routine workflows on the OEIS for unparalleled metabolite and isotopic coverage (<0.01 – 100% enrichment), 2) Examine plasma, urine and faecal samples from our 13C pea study archive by OEIS to identify and quantify the 13C metabolite landscape 3) from existing microbiome data analysis of these samples, determine microbial community structure-function relationships in vivo.

Fit to the scope of this opportunity: This project will provide proof of principle of new methodologies to Understanding the rules of life by using cutting-edge technology to provide new quantitative insights into the molecular signatures that connect the diet, the function of the microbiome and host tissues. Understanding the complexity and quantifying this signalling superhighway within organs, between organs and over time is fundamental to engineering new solutions for health (optimised and personalised diets, targeted molecular interventions (doi:10.1136/gutjnl-2014-307913) or new biotherapeutics to beneficially manipulate microbial activity).

This approach will challenge current paradigms because it moves beyond models by exploiting new technologies to give detailed mechanistic insight in natural human experiments. The OEIS is Transformative Technology and SUERC through a partnership with TFS is the first UK Centre to establish this technology which facilitates a step-change in our capability to undertake complex SI studies in humans. The multidisciplinary nature of SUERC makes it the ideal place to Drive convergence and integration across disciplines, particularly between the physical and life sciences, to support the development and application of new technologies to drive bioscience discovery. In particular the OEIS will increase the sensitivity, speed, resolution, and non-invasive methods of analytical measuring technologies, opening new avenues for research on living systems which has applications right across UKRI remit.

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Funded by: UKRI BBSRC

First published: 13 October 2023