Reductive carboxylation supports redox homeostasis during anchorage-independent growth
Open accessAbstract
Cells receive growth and survival stimuli through their attachment to an extracellular matrix (ECM). Overcoming the addiction to ECM-induced signals is required for anchorage-independent growth, a property of most malignant cells. Detachment from ECM is associated with enhanced production of reactive oxygen species (ROS) owing to altered glucose metabolism. Here we identify an unconventional pathway that supports redox homeostasis and growth during adaptation to anchorage independence. We observed that detachment from monolayer culture and growth as anchorage-independent tumour spheroids was accompanied by changes in both glucose and glutamine metabolism. Specifically, oxidation of both nutrients was suppressed in spheroids, whereas reductive formation of citrate from glutamine was enhanced. Reductive glutamine metabolism was highly dependent on cytosolic isocitrate dehydrogenase-1 (IDH1), because the activity was suppressed in cells homozygous null for IDH1 or treated with an IDH1 inhibitor. This activity occurred in absence of hypoxia, a well-known inducer of reductive metabolism. Rather, IDH1 mitigated mitochondrial ROS in spheroids, and suppressing IDH1 reduced spheroid growth through a mechanism requiring mitochondrial ROS. Isotope tracing revealed that in spheroids, isocitrate/citrate produced reductively in the cytosol could enter the mitochondria and participate in oxidative metabolism, including oxidation by IDH2. This generates NADPH in the mitochondria, enabling cells to mitigate mitochondrial ROS and maximize growth. Neither IDH1 nor IDH2 was necessary for monolayer growth, but deleting either one enhanced mitochondrial ROS and reduced spheroid size, as did deletion of the mitochondrial citrate transporter protein. Together, the data indicate that adaptation to anchorage independence requires a fundamental change in citrate metabolism, initiated by IDH1-dependent reductive carboxylation and culminating in suppression of mitochondrial ROS.
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Cancer cells metabolised glutamine differently when cultured as monolayers or spheroids or when transplanted as tumours.
“cancer cells metabolised glutamine differently when they were cultured as monolayers or spheroids [77] or when they were transplanted as tumours [78].”
From HIF-1-Independent Mechanisms Regulating Metabolic Adaptation in Hypoxic Cancer Cells (2021) · cites this paper as [77]
IDH1-dependent reductive glutamine metabolism produces NADPH, which decreases mitochondrial ROS during anchorage-independent growth.
“In addition, IDH1-dependent reductive glutamine metabolism produces NADPH, which decreases mitochondrial ROS during anchorage-independent growth64.”
From Glutamine reliance in cell metabolism (2020) · cites this paper as [64]
Citrate is generated by citrate synthase or through the reduction of α-ketoglutarate by isocitrate dehydrogenase.
“Citrate is generated by citrate synthase (CS) or through the reduction of α-ketoglutarate by isocitrate dehydrogenase (IDH).79–81”
From The multifaceted contributions of mitochondria to cellular metabolism (2018) · cites this paper as [81]
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- Authors
- Jiang, L., Shestov, A. A., Swain, P., Yang, C., Parker, S. J., Wang, Q. A., Terada, L. S., Adams, N. D., McCabe, M. T., Pietrak, B., Schmidt, S., Metallo, C. M., Dranka, B. P., Schwartz, B., & DeBerardinis, R. J.
- Affiliations
- Children's Medical Center (US)
- University of Pennsylvania (US)
- University of California San Diego (US)
- The University of Texas Southwestern Medical Center (US)
- GlaxoSmithKline (United States) (US)
- Corresponding author
- DeBerardinis, Ralph J.
- ORCID
- Venue
- Nature
- Published
- 2016-04
- Volume
- 532
- Issue
- 7598
- Pages
- 255-258
- ISSN
- 0028-0836 (ISSN-L)
- 1476-4687
- PMC
- PMC4860952
- OpenAlex
- W2312291090
- Cited by
- 582
- References
- 29
- Citations by year
- 2026: 36 · 2025: 33 · 2024: 50 · 2023: 56 · 2022: 77 · 2021: 71 · 2020: 72 · 2019: 60 · 2018: 54 · 2017: 48 · 2016: 23
- FWCI
- 25.69
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- RePEc: Research Papers in Economics (submittedVersion)
- eScholarship (California Digital Library) (submittedVersion)
- Funders
- National Cancer Institute (R01CA188652, R01CA157996)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Cancer, Hypoxia, and Metabolism
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Cancer and biochemical research
- Keywords
- reductive carboxylation, anchorage-independent growth, redox homeostasis, isocitrate dehydrogenase, IDH1, IDH2, citrate, mitochondrial reactive oxygen species, NADPH, tumor spheroids, oxidative stress, cancer metabolism, extracellular matrix detachment, isotope tracing, metabolic reprogramming
- MeSH terms
- Cell Adhesion, Contact Inhibition, Cytosol, Extracellular Matrix, Glucose, Glutamine, Homeostasis, Humans, Isocitrate Dehydrogenase, Isocitrates, Mitochondria, NADP, Neoplasms, Oxidation-Reduction, Cell Hypoxia, Reactive Oxygen Species, Oxidative Stress, Glutamic Acid, Spheroids, Cellular, Citric Acid, Cell Line, Tumor, Cell Proliferation
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