Reductive carboxylation supports growth in tumour cells with defective mitochondria (original) (raw)

• Letter
• Published: 20 November 2011
• William W. Wheaton2,3,
• Eunsook S. Jin4,5,
• Pei-Hsuan Chen1,
• Lucas B. Sullivan2,3,
• Tzuling Cheng1,
• Youfeng Yang6,
• W. Marston Linehan6,
• Navdeep S. Chandel2,3 &
• …
• Ralph J. DeBerardinis1,7,8

Nature volume 481, pages 385–388 (2012)Cite this article

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Abstract

Mitochondrial metabolism provides precursors to build macromolecules in growing cancer cells1,2. In normally functioning tumour cell mitochondria, oxidative metabolism of glucose- and glutamine-derived carbon produces citrate and acetyl-coenzyme A for lipid synthesis, which is required for tumorigenesis3. Yet some tumours harbour mutations in the citric acid cycle (CAC) or electron transport chain (ETC) that disable normal oxidative mitochondrial function4,5,6,7, and it is unknown how cells from such tumours generate precursors for macromolecular synthesis. Here we show that tumour cells with defective mitochondria use glutamine-dependent reductive carboxylation rather than oxidative metabolism as the major pathway of citrate formation. This pathway uses mitochondrial and cytosolic isoforms of NADP+/NADPH-dependent isocitrate dehydrogenase, and subsequent metabolism of glutamine-derived citrate provides both the acetyl-coenzyme A for lipid synthesis and the four-carbon intermediates needed to produce the remaining CAC metabolites and related macromolecular precursors. This reductive, glutamine-dependent pathway is the dominant mode of metabolism in rapidly growing malignant cells containing mutations in complex I or complex III of the ETC, in patient-derived renal carcinoma cells with mutations in fumarate hydratase, and in cells with normal mitochondria subjected to acute pharmacological ETC inhibition. Our findings reveal the novel induction of a versatile glutamine-dependent pathway that reverses many of the reactions of the canonical CAC, supports tumour cell growth, and explains how cells generate pools of CAC intermediates in the face of impaired mitochondrial metabolism.

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References

1. Vander Heiden, M. G., Cantley, L. C. & Thompson, C. B. Understanding the Warburg effect: the metabolic requirements of cell proliferation. Science 324, 1029–1033 (2009)
   Article ADS CAS Google Scholar
2. Gao, P. et al. c-Myc suppression of miR-23a/b enhances mitochondrial glutaminase expression and glutamine metabolism. Nature 458, 762–765 (2009)
   Article ADS CAS Google Scholar
3. DeBerardinis, R. J. et al. Beyond aerobic glycolysis: transformed cells can engage in glutamine metabolism that exceeds the requirement for protein and nucleotide synthesis. Proc. Natl Acad. Sci. USA 104, 19345–19350 (2007)
   Article ADS CAS Google Scholar
4. Linehan, W. M., Srinivasan, R. & Schmidt, L. S. The genetic basis of kidney cancer: a metabolic disease. Nature Rev. Urol. 7, 277–285 (2010)
   Article CAS Google Scholar
5. Baysal, B. E. et al. Mutations in SDHD, a mitochondrial complex II gene, in hereditary paraganglioma. Science 287, 848–851 (2000)
   Article ADS CAS Google Scholar
6. Tomlinson, I. P. et al. Germline mutations in FH predispose to dominantly inherited uterine fibroids, skin leiomyomata and papillary renal cell cancer. Nature Genet. 30, 406–410 (2002)
   Article CAS Google Scholar
7. Hao, H. X. et al. SDH5, a gene required for flavination of succinate dehydrogenase, is mutated in paraganglioma. Science 325, 1139–1142 (2009)
   Article ADS CAS Google Scholar
8. Rana, M., de Coo, I., Diaz, F., Smeets, H. & Moraes, C. T. An out-of-frame cytochrome b gene deletion from a patient with parkinsonism is associated with impaired complex III assembly and an increase in free radical production. Ann. Neurol. 48, 774–781 (2000)
   Article CAS Google Scholar
9. Weinberg, F. et al. Mitochondrial metabolism and ROS generation are essential for Kras-mediated tumorigenicity. Proc. Natl Acad. Sci. USA 107, 8788–8793 (2010)
   Article ADS CAS Google Scholar
10. Reitzer, L. J., Wice, B. M. & Kennell, D. Evidence that glutamine, not sugar, is the major energy source for cultured HeLa cells. J. Biol. Chem. 254, 2669–2676 (1979)
    CAS PubMed Google Scholar
11. Cheng, T. et al. Pyruvate carboxylase is required for glutamine-independent growth of tumor cells. Proc. Natl Acad. Sci. USA 108, 8674–8679 (2011)
    Article ADS CAS Google Scholar
12. Des Rosiers, C., Fernandez, C. A., David, F. & Brunengraber, H. Reversibility of the mitochondrial isocitrate dehydrogenase reaction in the perfused rat liver. Evidence from isotopomer analysis of citric acid cycle intermediates. J. Biol. Chem. 269, 27179–27182 (1994)
    CAS PubMed Google Scholar
13. Ward, P. S. et al. The common feature of leukemia-associated IDH1 and IDH2 mutations is a neomorphic enzyme activity converting alpha-ketoglutarate to 2-hydroxyglutarate. Cancer Cell 17, 225–234 (2010)
    Article CAS Google Scholar
14. Frezza, C., Pollard, P. J. & Gottlieb, E. Inborn and acquired metabolic defects in cancer. J. Mol. Med. 89, 213–220 (2011)
    Article CAS Google Scholar
15. Yang, Y. et al. UOK 262 cell line, fumarate hydratase deficient (FH-/FH-) hereditary leiomyomatosis renal cell carcinoma: in vitro and in vivo model of an aberrant energy metabolic pathway in human cancer. Cancer Genet. Cytogenet. 196, 45–55 (2010)
    Article CAS Google Scholar
16. Rossignol, R. et al. Energy substrate modulates mitochondrial structure and oxidative capacity in cancer cells. Cancer Res. 64, 985–993 (2004)
    Article CAS Google Scholar
17. El-Mir, M. Y. et al. Dimethylbiguanide inhibits cell respiration via an indirect effect targeted on the respiratory chain complex I. J. Biol. Chem. 275, 223–228 (2000)
    Article CAS Google Scholar
18. Yoo, H., Antoniewicz, M. R., Stephanopoulos, G. & Kelleher, J. K. Quantifying reductive carboxylation flux of glutamine to lipid in a brown adipocyte cell line. J. Biol. Chem. 283, 20621–20627 (2008)
    Article CAS Google Scholar
19. Comte, B., Vincent, G., Bouchard, B., Benderdour, M. & Des Rosiers, C. Reverse flux through cardiac NADP(+)-isocitrate dehydrogenase under normoxia and ischemia. Am. J. Physiol. Heart Circ. Physiol. 283, H1505–H1514 (2002)
    Article CAS Google Scholar
20. Lemons, J. M. et al. Quiescent fibroblasts exhibit high metabolic activity. PLoS Biol. 8, e1000514 (2010)
    Article Google Scholar
21. Olszewski, K. L. et al. Branched tricarboxylic acid metabolism in Plasmodium falciparum . Nature 466, 774–778 (2010)
    Article ADS CAS Google Scholar
22. DeBerardinis, R. J. & Cheng, T. Q's next: the diverse functions of glutamine in metabolism, cell biology and cancer. Oncogene 29, 313–324 (2009)
    Article Google Scholar
23. Brunelle, J. K. et al. Oxygen sensing requires mitochondrial ROS but not oxidative phosphorylation. Cell Metab. 1, 409–414 (2005)
    Article CAS Google Scholar
24. Yang, C. et al. Glioblastoma cells require glutamate dehydrogenase to survive impairments of glucose metabolism or Akt signaling. Cancer Res. 69, 7986–7993 (2009)
    Article CAS Google Scholar
25. Fernandez, C. A., Des Rosiers, C., Previs, S. F., David, F. & Brunengraber, H. Correction of 13C mass isotopomer distributions for natural stable isotope abundance. J. Mass Spectrom. 31, 255–262 (1996)
    Article ADS CAS Google Scholar
26. Frezza, C., Cipolat, S. & Scorrano, L. Organelle isolation: functional mitochondria from mouse liver, muscle and cultured fibroblasts. Nature Protocols 2, 287–295 (2007)
    Article CAS Google Scholar

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Acknowledgements

K. Uyeda and members of the DeBerardinis and Chandel laboratories provided critical assessment of the data, and J. Sudderth and C. Yang provided experimental assistance. We thank C. Moraes and I.F.M. de Coo for the WT 143B and CYTB 143B cell lines, I.E. Scheffler for CCL16-B2 cells and T. Yagi for CCL16-NDI1 cells. This work was supported by grants to R.J.D. from the NIH (K08DK072565 and R01CA157996), the Cancer Prevention and Research Institute of Texas (CPRIT, HIRP100437) and the Robert A. Welch Foundation (I1733); to N.S.C. from the NIH (R01CA123067), the LUNGevity Foundation and a Consortium of Independent Lung Health Organizations convened by Respiratory Health Association of Metropolitan Chicago; and to E.S.J. by an NIH grant (DK078933). The work was also supported by the Intramural Research Program of the NIH, National Cancer Institute Center for Cancer Research and by NIH grant RR02584. NIH training grants supported A.R.M. (5T32GM083831), W.W.W. (T32CA009560) and L.B.S. (T32GM008061). T.C. was supported by a CPRIT training grant.

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Authors and Affiliations

1. Department of Pediatrics, University of Texas – Southwestern Medical Center at Dallas, Dallas, 75390-9063, Texas, USA
   Andrew R. Mullen, Pei-Hsuan Chen, Tzuling Cheng & Ralph J. DeBerardinis
2. Department of Medicine, Northwestern University, Chicago, 60611-3008, Illinois, USA
   William W. Wheaton, Lucas B. Sullivan & Navdeep S. Chandel
3. Department of Cell and Molecular Biology, Northwestern University, Chicago, 60611-3008, Illinois, USA
   William W. Wheaton, Lucas B. Sullivan & Navdeep S. Chandel
4. Department of Internal Medicine, University of Texas – Southwestern Medical Center at Dallas, Dallas, 75390-9063, Texas, USA
   Eunsook S. Jin
5. Advanced Imaging Research Center, University of Texas – Southwestern Medical Center at Dallas, Dallas, 75390-8568, Texas, USA
   Eunsook S. Jin
6. Urologic Oncology Branch, National Cancer Institute, Bethesda, 20892, Maryland, USA
   Youfeng Yang & W. Marston Linehan
7. McDermott Center for Human Growth and Development, University of Texas – Southwestern Medical Center at Dallas, Dallas, 75390-8591, Texas, USA
   Ralph J. DeBerardinis
8. Harold C. Simmons Comprehensive Cancer Center, University of Texas – Southwestern Medical Center at Dallas, Dallas, 75235-5303, Texas, USA
   Ralph J. DeBerardinis

Authors

1. Andrew R. Mullen
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2. William W. Wheaton
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3. Eunsook S. Jin
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4. Pei-Hsuan Chen
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5. Lucas B. Sullivan
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6. Tzuling Cheng
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7. Youfeng Yang
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8. W. Marston Linehan
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9. Navdeep S. Chandel
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10. Ralph J. DeBerardinis
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Contributions

A.R.M., W.W.W., N.S.C. and R.J.D. designed the research. A.R.M., W.W.W., L.B.S., E.S.J., T.C. and P.-H.C. performed the experiments. A.R.M., W.W.W., L.B.S., E.S.J., P.-H.C., T.C., N.S.C. and R.J.D. analysed the data. Y.Y. and W.M.L. provided the FH-deficient (UOK262) cells. A.R.M., N.S.C. and R.J.D. wrote the paper.

Corresponding authors

Correspondence toNavdeep S. Chandel or Ralph J. DeBerardinis.

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The authors declare no competing financial interests.

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Mullen, A., Wheaton, W., Jin, E. et al. Reductive carboxylation supports growth in tumour cells with defective mitochondria.Nature 481, 385–388 (2012). https://doi.org/10.1038/nature10642

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• Received: 13 June 2011
• Accepted: 13 October 2011
• Published: 20 November 2011
• Issue Date: 19 January 2012
• DOI: https://doi.org/10.1038/nature10642

Editorial Summary

Alternative route to fatty acids

Oxidative metabolism of glucose has long been considered to be the major provider of carbon for lipid synthesis in animal cells. Two papers in this issue of Nature demonstrate that reductive carboxylation of glutamine is an alternative. Metallo et al. show that various normal and cancerous human cell lines proliferating in hypoxic conditions produce the acetyl-coenzyme A required as a precursor for fatty acid synthesis by the reductive metabolism of glutamine-derived α-ketoglutarate through a pathway requiring isocitrate dehydrogenase 1. Mullen et al. show that tumour cells with defective mitochondria use glutamine-dependent reductive carboxylation as the major pathway of citrate formation. As well as adding a new dimension to our understanding of cell carbohydrate metabolism, this work suggests that there may be potential therapeutic targets along the reductive carboxylation and glutamine catabolic pathways that could prevent hypoxic tumour growth.