Spatial-fluxomics provides a subcellular-compartmentalized view of reductive glutamine metabolism in cancer cells
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The inability to inspect metabolic activities within subcellular compartments has been a major barrier to our understanding of eukaryotic cell metabolism. Here, we describe a spatial-fluxomics approach for inferring metabolic fluxes in mitochondria and cytosol under physiological conditions, combining isotope tracing, rapid subcellular fractionation, LC-MS-based metabolomics, computational deconvolution, and metabolic network modeling. Applied to study reductive glutamine metabolism in cancer cells, shown to mediate fatty acid biosynthesis under hypoxia and defective mitochondria, we find a previously unappreciated role of reductive IDH1 as the sole net contributor of carbons to fatty acid biosynthesis under standard normoxic conditions in HeLa cells. In murine cells with defective SDH, we find that reductive biosynthesis of citrate in mitochondria is followed by a reversed CS activity, suggesting a new route for supporting pyrimidine biosynthesis. We expect this spatial-fluxomics approach to be a highly useful tool for elucidating the role of metabolic dysfunction in human disease.
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CS reversibility has been observed in mammalian cells in the context of SDH deficiency, where reductive glutamine metabolism produces citrate and citrate-derived OAA that can sustain biosynthesis.
“Even CS may reverse: CS reversibility, which occurs in prokaryotes under high CO2 conditions (see aforementioned), has been observed in mammalian cells in the context of SDH deficiency in which reductive glutamine metabolism produces citrate and, strikingly, citrate-derived OAA that can be used to sustain biosynthesis (132).”
From Regulation and function of the mammalian tricarboxylic acid cycle (2023) · cites this paper as [132]
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Regulation and function of the mammalian tricarboxylic acid cycle
Arnold, P. K., & Finley, L. W. S. (2023) · Journal of Biological Chemistry · doi:10.1016/j.jbc.2022.102838 · Cited by 535 · see citing works
“(2023) 299(2) 102838 observed in mammalian cells in the context of SDH deficiency in which reductive glutamine metabolism produces citrate and, strikingly, citrate-derived OAA that can be used to sustain biosynthesis (132).”
BackgroundIns and outs of the TCA cycle: the central role of anaplerosis
Inigo, M., Deja, S., & Burgess, S. C. (2021) · Annual Review of Nutrition · doi:10.1146/annurev-nutr-120420-025558 · Cited by 189 · see citing works
- Authors
- Lee, W. D., Mukha, D., Aizenshtein, E., & Shlomi, T.
- Affiliations
- Technion – Israel Institute of Technology (IL)
- Corresponding author
- Shlomi, Tomer
- Venue
- Nature Communications
- Published
- 2019-03-22
- Volume
- 10
- Issue
- 1
- Pages
- 1351-1351
- ISSN
- 2041-1723 (ISSN-L)
- PMC
- PMC6430770
- OpenAlex
- W2924911015
- Cited by
- 70
- References
- 68
- Citations by year
- 2026: 7 · 2025: 2 · 2024: 6 · 2023: 12 · 2022: 18 · 2021: 10 · 2020: 11 · 2019: 4
- FWCI
- 3.39
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- Nature Communications (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- RePEc: Research Papers in Economics (submittedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- H2020 European Research Council (714738)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Cancer, Hypoxia, and Metabolism
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Metabolomics and Mass Spectrometry Studies
- Keywords
- cancer metabolism, subcellular compartmentalization, isotope tracing, subcellular fractionation, fatty acid synthesis, hypoxia, pyrimidine synthesis, succinate dehydrogenase deficiency, HeLa cells, metabolic network modeling
- MeSH terms
- Animals, Carbon Isotopes, Cell Compartmentation, Citrate (si)-Synthase, Citric Acid Cycle, Cytosol, Glutamine, HeLa Cells, Humans, Isocitrate Dehydrogenase, Mitochondria, Neoplasms, Subcellular Fractions, Succinate Dehydrogenase, Cell Hypoxia, Citric Acid, Mice, Metabolome, Metabolic Flux Analysis
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