Glioma Cells with the IDH1 Mutation Modulate Metabolic Fractional Flux through Pyruvate Carboxylase

Background Over 70% of low-grade gliomas carry a heterozygous R132H mutation in the gene coding for isocitrate dehydrogenase 1 (IDH1). This confers the enzyme with the novel ability to convert α-ketoglutarate to 2-hydroxyglutarate, ultimately leading to tumorigenesis. The major source of 2-hydroxyglutarate production is glutamine, which, in cancer, is also a source for tricarboxylic acid cycle (TCA) anaplerosis. An alternate source of anaplerosis is pyruvate flux via pyruvate carboxylase (PC), which is a common pathway in normal astrocytes. The goal of this study was to determine whether PC serves as a source of TCA anaplerosis in IDH1 mutant cells wherein glutamine is used for 2-hydroxyglutarate production. Methods Immortalized normal human astrocytes engineered to express heterozygous mutant IDH1 or wild-type IDH1 were investigated. Flux of pyruvate via PC and via pyruvate dehydrogenase (PDH) was determined by using magnetic resonance spectroscopy to probe the labeling of [2-13C]glucose-derived 13C-labeled glutamate and glutamine. Activity assays, RT-PCR and western blotting were used to probe the expression and activity of relevant enzymes. The Cancer Genome Atlas (TCGA) data was analyzed to assess the expression of enzymes in human glioma samples. Results Compared to wild-type cells, mutant IDH1 cells significantly increased fractional flux through PC. This was associated with a significant increase in PC activity and expression. Concurrently, PDH activity significantly decreased, likely mediated by significantly increased inhibitory PDH phosphorylation by PDH kinase 3. Consistent with the observation in cells, analysis of TCGA data indicated a significant increase in PC expression in mutant IDH-expressing human glioma samples compared to wild-type IDH. Conclusions Our findings suggest that changes in PC and PDH may be an important part of cellular adaptation to the IDH1 mutation and may serve as potential therapeutic targets.

[1]  S. Weiss,et al.  Lactate dehydrogenase A silencing in IDH mutant gliomas. , 2014, Neuro-oncology.

[2]  R. Deberardinis,et al.  Simultaneous Steady-state and Dynamic 13C NMR Can Differentiate Alternative Routes of Pyruvate Metabolism in Living Cancer Cells* , 2014, The Journal of Biological Chemistry.

[3]  L. Deangelis,et al.  Glioblastoma and other malignant gliomas: a clinical review. , 2013, JAMA.

[4]  L. Deangelis,et al.  Glioblastoma and Other Malignant Gliomas , 2013 .

[5]  D. Haussler,et al.  The Somatic Genomic Landscape of Glioblastoma , 2013, Cell.

[6]  Daniel B Vigneron,et al.  Non-invasive in vivo assessment of IDH1 mutational status in glioma , 2013, Nature Communications.

[7]  T. Mak,et al.  Oncogenic isocitrate dehydrogenase mutations: mechanisms, models, and clinical opportunities. , 2013, Cancer discovery.

[8]  A. Schousboe,et al.  Astrocytic Control of Biosynthesis and Turnover of the Neurotransmitters Glutamate and GABA , 2013, Front. Endocrinol..

[9]  Boris Ratnikov,et al.  Glutamine‐fueled mitochondrial metabolism is decoupled from glycolysis in melanoma , 2012, Pigment cell & melanoma research.

[10]  W. Marston Linehan,et al.  Targeting Cancer Metabolism , 2012, Clinical Cancer Research.

[11]  Dania Daye,et al.  Metabolic reprogramming in cancer: unraveling the role of glutamine in tumorigenesis. , 2012, Seminars in cell & developmental biology.

[12]  A. Schousboe,et al.  Direct measurement of backflux between oxaloacetate and fumarate following pyruvate carboxylation , 2012, Glia.

[13]  Leif Hertz,et al.  Astrocytic energy metabolism and glutamate formation--relevance for 13C-NMR spectroscopy and importance of cytosolic/mitochondrial trafficking. , 2011, Magnetic resonance imaging.

[14]  Dinesh Rakheja,et al.  2-hydroxyglutarate detection by magnetic resonance spectroscopy in IDH-mutated glioma patients , 2011, Nature Medicine.

[15]  M. V. Heiden,et al.  Targeting cancer metabolism: a therapeutic window opens , 2011, Nature Reviews Drug Discovery.

[16]  Ralph J DeBerardinis,et al.  Role of Glutamine in Cancer: Therapeutic and Imaging Implications , 2011, The Journal of Nuclear Medicine.

[17]  R. Deberardinis,et al.  Pyruvate carboxylase is required for glutamine-independent growth of tumor cells , 2011, Proceedings of the National Academy of Sciences.

[18]  G. Riggins,et al.  Inhibition of glutaminase preferentially slows growth of glioma cells with mutant IDH1. , 2010, Cancer research.

[19]  E. Gottlieb,et al.  Targeting metabolic transformation for cancer therapy , 2010, Nature Reviews Cancer.

[20]  L. Liau,et al.  Cancer-associated IDH1 mutations produce 2-hydroxyglutarate , 2009, Nature.

[21]  L. Cantley,et al.  Understanding the Warburg Effect: The Metabolic Requirements of Cell Proliferation , 2009, Science.

[22]  C. Thompson,et al.  Metabolic enzymes as oncogenes or tumor suppressors. , 2009, The New England journal of medicine.

[23]  Andrey Korshunov,et al.  Analysis of the IDH1 codon 132 mutation in brain tumors , 2008, Acta Neuropathologica.

[24]  D. Brat,et al.  Diagnosis of malignant glioma: role of neuropathology , 2008, Journal of Neuro-Oncology.

[25]  R. Deberardinis,et al.  Beyond aerobic glycolysis: Transformed cells can engage in glutamine metabolism that exceeds the requirement for protein and nucleotide synthesis , 2007, Proceedings of the National Academy of Sciences.

[26]  D. Botstein,et al.  Gene expression profiling reveals molecularly and clinically distinct subtypes of glioblastoma multiforme. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[27]  R. Gruetter,et al.  Neuroglial Metabolism in the Awake Rat Brain: CO2 Fixation Increases with Brain Activity , 2004, The Journal of Neuroscience.

[28]  J. Griffiths,et al.  Magnetic resonance spectroscopic pharmacodynamic markers of the heat shock protein 90 inhibitor 17-allylamino,17-demethoxygeldanamycin (17AAG) in human colon cancer models. , 2003, Journal of the National Cancer Institute.

[29]  M. Holness,et al.  Recent advances in mechanisms regulating glucose oxidation at the level of the pyruvate dehydrogenase complex by PDKs. , 2003, American journal of physiology. Endocrinology and metabolism.

[30]  L. Sokoloff,et al.  Dichloroacetate effects on glucose and lactate oxidation by neurons and astroglia in vitro and on glucose utilization by brain in vivo , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[31]  S. Kalhan,et al.  The Key Role of Anaplerosis and Cataplerosis for Citric Acid Cycle Function* , 2002, The Journal of Biological Chemistry.

[32]  K. Petersen,et al.  Astroglial Contribution to Brain Energy Metabolism in Humans Revealed by 13C Nuclear Magnetic Resonance Spectroscopy: Elucidation of the Dominant Pathway for Neurotransmitter Glutamate Repletion and Measurement of Astrocytic Oxidative Metabolism , 2002, The Journal of Neuroscience.

[33]  K. Aldape,et al.  Formation of intracranial tumors by genetically modified human astrocytes defines four pathways critical in the development of human anaplastic astrocytoma. , 2001, Cancer research.

[34]  H. Degani,et al.  Simultaneous extraction of cellular lipids and water‐soluble metabolites: Evaluation by NMR spectroscopy , 1996, Magnetic resonance in medicine.

[35]  R. Shank,et al.  Pyruvate car☐ylase: an astrocyte-specific enzyme implicated in the replenishment of amino acid neurotransmitter pools , 1985, Brain Research.

[36]  L. Hertz,et al.  Abnormal metabolic response to excess potassium in astrocytes from the Jimpy mouse, a convulsing neurological mutant , 1980, Brain Research.

[37]  L. Hertz,et al.  Polarographic measurement of oxygen uptake by astrocytes in primary cultures using the tissue-culture flask as the respirometer chamber , 1979, In Vitro.

[38]  I. Siegel.  A clinical review , 1978 .

[39]  R. Deberardinis,et al.  2-hydroxyglutarate detection by magnetic resonance spectroscopy in subjects with IDH-mutated gliomas , 2015 .

[40]  F. Fonnum,et al.  NMR spectroscopic studies of 13C acetate and 13C glucose metabolism in neocortical astrocytes: evidence for mitochondrial heterogeneity. , 1993, Developmental neuroscience.

[41]  R. Denton,et al.  Regulation of pyruvate metabolism in mammalian tissues. , 1979, Essays in biochemistry.

[42]  Denton Rm,et al.  Regulation of pyruvate metabolism in mammalian tissues. , 1979 .