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.
This is a preview of subscription content, access via your institution
Open Access articles citing this article.
Scientific Reports Open Access 07 November 2022
Journal of Experimental & Clinical Cancer Research Open Access 24 September 2022
Cell Death & Differentiation Open Access 13 July 2022
Subscribe to Journal
Get full journal access for 1 year
only $3.90 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Tax calculation will be finalised during checkout.
Get time limited or full article access on ReadCube.
All prices are NET prices.
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)
Gao, P. et al. c-Myc suppression of miR-23a/b enhances mitochondrial glutaminase expression and glutamine metabolism. Nature 458, 762–765 (2009)
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)
Linehan, W. M., Srinivasan, R. & Schmidt, L. S. The genetic basis of kidney cancer: a metabolic disease. Nature Rev. Urol. 7, 277–285 (2010)
Baysal, B. E. et al. Mutations in SDHD, a mitochondrial complex II gene, in hereditary paraganglioma. Science 287, 848–851 (2000)
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)
Hao, H. X. et al. SDH5, a gene required for flavination of succinate dehydrogenase, is mutated in paraganglioma. Science 325, 1139–1142 (2009)
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)
Weinberg, F. et al. Mitochondrial metabolism and ROS generation are essential for Kras-mediated tumorigenicity. Proc. Natl Acad. Sci. USA 107, 8788–8793 (2010)
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)
Cheng, T. et al. Pyruvate carboxylase is required for glutamine-independent growth of tumor cells. Proc. Natl Acad. Sci. USA 108, 8674–8679 (2011)
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)
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)
Frezza, C., Pollard, P. J. & Gottlieb, E. Inborn and acquired metabolic defects in cancer. J. Mol. Med. 89, 213–220 (2011)
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)
Rossignol, R. et al. Energy substrate modulates mitochondrial structure and oxidative capacity in cancer cells. Cancer Res. 64, 985–993 (2004)
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)
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)
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)
Lemons, J. M. et al. Quiescent fibroblasts exhibit high metabolic activity. PLoS Biol. 8, e1000514 (2010)
Olszewski, K. L. et al. Branched tricarboxylic acid metabolism in Plasmodium falciparum . Nature 466, 774–778 (2010)
DeBerardinis, R. J. & Cheng, T. Q's next: the diverse functions of glutamine in metabolism, cell biology and cancer. Oncogene 29, 313–324 (2009)
Brunelle, J. K. et al. Oxygen sensing requires mitochondrial ROS but not oxidative phosphorylation. Cell Metab. 1, 409–414 (2005)
Yang, C. et al. Glioblastoma cells require glutamate dehydrogenase to survive impairments of glucose metabolism or Akt signaling. Cancer Res. 69, 7986–7993 (2009)
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)
Frezza, C., Cipolat, S. & Scorrano, L. Organelle isolation: functional mitochondria from mouse liver, muscle and cultured fibroblasts. Nature Protocols 2, 287–295 (2007)
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.
The authors declare no competing financial interests.
About this article
Cite this article
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
This article is cited by
Journal of Experimental & Clinical Cancer Research (2022)
Cancer & Metabolism (2022)
British Journal of Cancer (2022)
Non-oxidative pentose phosphate pathway controls regulatory T cell function by integrating metabolism and epigenetics
Nature Metabolism (2022)
Mitochondria preserve an autarkic one-carbon cycle to confer growth-independent cancer cell migration and metastasis
Nature Communications (2022)