A framework for tracer-based metabolism in mammalian cells by NMR

Raquel Saborano; Zuhal Eraslan; Jennie Roberts; Farhat L. Khanim; Patricia F. Lalor; Michelle A.C. Reed; Ulrich L. Günther

doi:10.1038/s41598-018-37525-3

A framework for tracer-based metabolism in mammalian cells by NMR

Raquel Saborano, Zuhal Eraslan, Jennie Roberts, Farhat L. Khanim, Patricia F. Lalor, Michelle A.C. Reed, Ulrich L. Günther^*

^*Corresponding author for this work

Institute of Chemistry and Metabolomics

University of Birmingham

34 Citations (Scopus)

Abstract

Metabolism changes extensively during the normal proliferation and differentiation of mammalian cells, and in cancer and inflammatory diseases. Since changes in the metabolic network reflect interactions between genetic, epigenetic and environmental changes, it is helpful to study the flow of label from isotopically labelled precursors into other metabolites rather than static metabolite levels. For this Nuclear Magnetic Resonance (NMR) spectroscopy is an attractive technique as it can quantify site-specific label incorporation. However, for applications using human cells and cell lines, the challenge is to optimize the process to maximize sensitivity and reproducibility. Here we present a new framework to analyze metabolism in mammalian cell lines and primary cells, covering the workflow from the preparation of cells to the acquisition and analysis of NMR spectra. We have applied this new approach in hematological and liver cancer cell lines and confirm the feasibility of tracer-based metabolism in primary liver cells.

Original language	English
Article number	2520
Journal	Scientific Reports
Volume	9
Issue number	1
ISSN	2045-2322
DOIs	https://doi.org/10.1038/s41598-018-37525-3
Publication status	Published - 01.12.2019

Funding

Much of the work presented is based on the FP7 Metaflux project (FP7-PEOPLE-ITN-2010–264780). We thank the EC for supporting ZE and RS within the EUROPOL-ITN project (H2020-MSCA-ITN-2014-642773), PhenoMeNal for supporting software development (H2020-EINFRA-2014-2-654241), and BBSRC for supporting JR in the context of a BBSRC CASE studentship. We are also grateful to the Wellcome Trust for supporting access to NMR instruments at the Henry Wellcome Building for Biomolecular NMR in Birmingham (grant number 208400/Z/17/Z). We are also grateful for access to NMR instrumentation at the BMRZ NMR centre in Frankfurt with support from INext, and thank Christian Richter for technical assistance. This report includes independent research funded by the NIHR Birmingham Biomedical Research Centre at the University Hospitals Birmingham NHS Foundation Trust and the University of Birmingham. The views expressed are those of the author(s) and not necessarily those of the NHS, the NIHR or the Department of Health.

Research Areas and Centers

Academic Focus: Center for Infection and Inflammation Research (ZIEL)

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1038/s41598-018-37525-3

Cite this

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title = "A framework for tracer-based metabolism in mammalian cells by NMR",

abstract = "Metabolism changes extensively during the normal proliferation and differentiation of mammalian cells, and in cancer and inflammatory diseases. Since changes in the metabolic network reflect interactions between genetic, epigenetic and environmental changes, it is helpful to study the flow of label from isotopically labelled precursors into other metabolites rather than static metabolite levels. For this Nuclear Magnetic Resonance (NMR) spectroscopy is an attractive technique as it can quantify site-specific label incorporation. However, for applications using human cells and cell lines, the challenge is to optimize the process to maximize sensitivity and reproducibility. Here we present a new framework to analyze metabolism in mammalian cell lines and primary cells, covering the workflow from the preparation of cells to the acquisition and analysis of NMR spectra. We have applied this new approach in hematological and liver cancer cell lines and confirm the feasibility of tracer-based metabolism in primary liver cells.",

author = "Raquel Saborano and Zuhal Eraslan and Jennie Roberts and Khanim, \{Farhat L.\} and Lalor, \{Patricia F.\} and Reed, \{Michelle A.C.\} and G{\"u}nther, \{Ulrich L.\}",

note = "Funding Information: Much of the work presented is based on the FP7 Metaflux project (FP7-PEOPLE-ITN-2010–264780). We thank the EC for supporting ZE and RS within the EUROPOL-ITN project (H2020-MSCA-ITN-2014-642773), PhenoMeNal for supporting software development (H2020-EINFRA-2014-2-654241), and BBSRC for supporting JR in the context of a BBSRC CASE studentship. We are also grateful to the Wellcome Trust for supporting access to NMR instruments at the Henry Wellcome Building for Biomolecular NMR in Birmingham (grant number 208400/Z/17/Z). We are also grateful for access to NMR instrumentation at the BMRZ NMR centre in Frankfurt with support from INext, and thank Christian Richter for technical assistance. This report includes independent research funded by the NIHR Birmingham Biomedical Research Centre at the University Hospitals Birmingham NHS Foundation Trust and the University of Birmingham. The views expressed are those of the author(s) and not necessarily those of the NHS, the NIHR or the Department of Health. Publisher Copyright: {\textcopyright} 2019, The Author(s). Copyright: Copyright 2019 Elsevier B.V., All rights reserved.",

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AU - Reed, Michelle A.C.

AU - Günther, Ulrich L.

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N2 - Metabolism changes extensively during the normal proliferation and differentiation of mammalian cells, and in cancer and inflammatory diseases. Since changes in the metabolic network reflect interactions between genetic, epigenetic and environmental changes, it is helpful to study the flow of label from isotopically labelled precursors into other metabolites rather than static metabolite levels. For this Nuclear Magnetic Resonance (NMR) spectroscopy is an attractive technique as it can quantify site-specific label incorporation. However, for applications using human cells and cell lines, the challenge is to optimize the process to maximize sensitivity and reproducibility. Here we present a new framework to analyze metabolism in mammalian cell lines and primary cells, covering the workflow from the preparation of cells to the acquisition and analysis of NMR spectra. We have applied this new approach in hematological and liver cancer cell lines and confirm the feasibility of tracer-based metabolism in primary liver cells.

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A framework for tracer-based metabolism in mammalian cells by NMR

Abstract

Funding

Research Areas and Centers

UN SDGs

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