Using DIMet for Differential Analysis of Labeled Metabolomics Data: A Step-by-step Guide Showcasing the Glioblastoma Metabolism

Stable-isotope resolved metabolomics (SIRM) is a powerful approach for characterizing metabolic states in cells and organisms. By incorporating isotopes, such as 13C, into substrates, researchers can trace reaction rates across specific metabolic pathways. Integrating metabolomics data with gene exp...

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Main Authors: Johanna Galvis, Joris Guyon, Thomas Daubon, Macha Nikolski
Format: Article
Language:English
Published: Bio-protocol LLC 2025-01-01
Series:Bio-Protocol
Online Access:https://bio-protocol.org/en/bpdetail?id=5168&type=0
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author Johanna Galvis
Joris Guyon
Thomas Daubon
Macha Nikolski
author_facet Johanna Galvis
Joris Guyon
Thomas Daubon
Macha Nikolski
author_sort Johanna Galvis
collection DOAJ
description Stable-isotope resolved metabolomics (SIRM) is a powerful approach for characterizing metabolic states in cells and organisms. By incorporating isotopes, such as 13C, into substrates, researchers can trace reaction rates across specific metabolic pathways. Integrating metabolomics data with gene expression profiles further enriches the analysis, as we demonstrated in our prior study on glioblastoma metabolic symbiosis. However, the bioinformatics tools for analyzing tracer metabolomics data have been limited. In this protocol, we encourage the researchers to use SIRM and transcriptomics data and to perform the downstream analysis using our software tool DIMet. Indeed, DIMet is the first comprehensive tool designed for the differential analysis of tracer metabolomics data, alongside its integration with transcriptomics data. DIMet facilitates the analysis of stable-isotope labeling and metabolic abundances, offering a streamlined approach to infer metabolic changes without requiring complex flux analysis. Its pathway-based "metabologram" visualizations effectively integrate metabolomics and transcriptomics data, offering a versatile platform capable of analyzing corrected tracer datasets across diverse systems, organisms, and isotopes. We provide detailed steps for sample preparation and data analysis using DIMet through its intuitive, web-based Galaxy interface. To showcase DIMet's capabilities, we analyzed LDHA/B knockout glioblastoma cell lines compared to controls. Accessible to all researchers through Galaxy, DIMet is free, user-friendly, and open source, making it a valuable resource for advancing metabolic research.
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spelling doaj-art-eed75ddafefe4a82b050e662b5815ac52025-02-07T08:16:38ZengBio-protocol LLCBio-Protocol2331-83252025-01-0115210.21769/BioProtoc.5168Using DIMet for Differential Analysis of Labeled Metabolomics Data: A Step-by-step Guide Showcasing the Glioblastoma MetabolismJohanna Galvis0Joris Guyon1Thomas Daubon2Macha Nikolski3University of Bordeaux, CNRS, IBGC UMR 5095, Bordeaux, FranceUniversity of Bordeaux, Bordeaux Bioinformatics Center CBiB, Bordeaux, FranceUniversity of Bordeaux, INSERM, BRIC, UMR1312, Bordeaux, FranceDepartment of Medical Pharmacology, CHU Bordeaux, Bordeaux, FranceUniversity of Bordeaux, CNRS, IBGC UMR 5095, Bordeaux, FranceUniversity of Bordeaux, CNRS, IBGC UMR 5095, Bordeaux, FranceUniversity of Bordeaux, Bordeaux Bioinformatics Center CBiB, Bordeaux, FranceStable-isotope resolved metabolomics (SIRM) is a powerful approach for characterizing metabolic states in cells and organisms. By incorporating isotopes, such as 13C, into substrates, researchers can trace reaction rates across specific metabolic pathways. Integrating metabolomics data with gene expression profiles further enriches the analysis, as we demonstrated in our prior study on glioblastoma metabolic symbiosis. However, the bioinformatics tools for analyzing tracer metabolomics data have been limited. In this protocol, we encourage the researchers to use SIRM and transcriptomics data and to perform the downstream analysis using our software tool DIMet. Indeed, DIMet is the first comprehensive tool designed for the differential analysis of tracer metabolomics data, alongside its integration with transcriptomics data. DIMet facilitates the analysis of stable-isotope labeling and metabolic abundances, offering a streamlined approach to infer metabolic changes without requiring complex flux analysis. Its pathway-based "metabologram" visualizations effectively integrate metabolomics and transcriptomics data, offering a versatile platform capable of analyzing corrected tracer datasets across diverse systems, organisms, and isotopes. We provide detailed steps for sample preparation and data analysis using DIMet through its intuitive, web-based Galaxy interface. To showcase DIMet's capabilities, we analyzed LDHA/B knockout glioblastoma cell lines compared to controls. Accessible to all researchers through Galaxy, DIMet is free, user-friendly, and open source, making it a valuable resource for advancing metabolic research.https://bio-protocol.org/en/bpdetail?id=5168&type=0
spellingShingle Johanna Galvis
Joris Guyon
Thomas Daubon
Macha Nikolski
Using DIMet for Differential Analysis of Labeled Metabolomics Data: A Step-by-step Guide Showcasing the Glioblastoma Metabolism
Bio-Protocol
title Using DIMet for Differential Analysis of Labeled Metabolomics Data: A Step-by-step Guide Showcasing the Glioblastoma Metabolism
title_full Using DIMet for Differential Analysis of Labeled Metabolomics Data: A Step-by-step Guide Showcasing the Glioblastoma Metabolism
title_fullStr Using DIMet for Differential Analysis of Labeled Metabolomics Data: A Step-by-step Guide Showcasing the Glioblastoma Metabolism
title_full_unstemmed Using DIMet for Differential Analysis of Labeled Metabolomics Data: A Step-by-step Guide Showcasing the Glioblastoma Metabolism
title_short Using DIMet for Differential Analysis of Labeled Metabolomics Data: A Step-by-step Guide Showcasing the Glioblastoma Metabolism
title_sort using dimet for differential analysis of labeled metabolomics data a step by step guide showcasing the glioblastoma metabolism
url https://bio-protocol.org/en/bpdetail?id=5168&type=0
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