Abstract
We describe a liquid chromatography-tandem mass spectrometry assay for measurement of d-2-hydroxyglutaric acid and l-2-hydroxyglutaric acid. These metabolites are increased in specific inborn errors of metabolism and are now recognized as oncometabolites. The measurement of d-2-hydroxyglutarate in peripheral blood may be used as a biomarker for screening and follow-up of patients with IDH-mutated acute myeloid leukemia.
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References
Struys EA, Verhoeven NM, Jansen EE et al (2006) Metabolism of gamma-hydroxybutyrate to d-2-hydroxyglutarate in mammals: further evidence for d-2-hydroxyglutarate transhydrogenase. Metabolism 55:353–358
Struys EA, Verhoeven NM, Ten Brink HJ et al (2005) Kinetic characterization of human hydroxyacid-oxoacid transhydrogenase: relevance to D-2-hydroxyglutaric and gamma-hydroxybutyric acidurias. J Inherit Metab Dis 28:921–930
Van Schaftingen E, Rzem R, Veiga-da-Cunha M (2009) L-2-Hydroxyglutaric aciduria, a disorder of metabolite repair. J Inherit Metab Dis 32:135–142
Intlekofer AM, Dematteo RG, Venneti S et al (2015) Hypoxia induces production of L-2-Hydroxyglutarate. Cell Metab 22:304–311
Oldham WM, Clish CB, Yang Y et al (2015) Hypoxia-mediated increases in L-2-hydroxyglutarate coordinate the metabolic response to reductive stress. Cell Metab 22:291–303
Steenweg ME, Jakobs C, Errami A et al (2010) An overview of L-2-hydroxyglutarate dehydrogenase gene (L2HGDH) variants: a genotype-phenotype study. Hum Mutat 31:380–390
Topcu M, Jobard F, Halliez S et al (2004) L-2-Hydroxyglutaric aciduria: identification of a mutant gene C14orf160, localized on chromosome 14q22.1. Hum Mol Genet 13:2803–2811
Rzem R, Veiga-da-Cunha M, Noel G et al (2004) A gene encoding a putative FAD-dependent L-2-hydroxyglutarate dehydrogenase is mutated in L-2-hydroxyglutaric aciduria. Proc Natl Acad Sci U S A 101:16849–16854
Struys EA (2006) D-2-Hydroxyglutaric aciduria: Unravelling the biochemical pathway and the genetic defect. J Inherit Metab Dis 29:21–29
Kranendijk M, Struys EA, Gibson KM et al (2009) Evidence for genetic heterogeneity in D-2-hydroxyglutaric aciduria. Hum Mutat 31:279–283
Struys EA, Korman SH, Salomons GS et al (2005) Mutations in phenotypically mild D-2-hydroxyglutaric aciduria. Ann Neurol 58:626–630
Struys EA, Salomons GS, Achouri Y et al (2005) Mutations in the D-2-hydroxyglutarate dehydrogenase gene cause D-2-hydroxyglutaric aciduria. Am J Hum Genet 76:358–360
Nota B, Struys EA, Pop A et al (2013) Deficiency in SLC25A1, encoding the mitochondrial citrate carrier, causes combined D-2- and L-2-hydroxyglutaric aciduria. Am J Hum Genet 92:627–631
Kranendijk M, Struys EA, van Schaftingen E et al (2010) IDH2 mutations in patients with D-2-hydroxyglutaric aciduria. Science 330:336
Parsons DW, Jones S, Zhang X et al (2008) An integrated genomic analysis of human glioblastoma multiforme. Science 321:1807–1812
Yan H, Parsons DW, Jin G et al (2009) IDH1 and IDH2 mutations in gliomas. N Engl J Med 360:765–773
Mardis ER, Ding L, Dooling DJ et al (2009) Recurring mutations found by sequencing an acute myeloid leukemia genome. N Engl J Med 361:1058–1066
Thol F, Damm F, Wagner K et al (2010) Prognostic impact of IDH2 mutations in cytogenetically normal acute myeloid leukemia. Blood 116:614–616
Abbas S, Lugthart S, Kavelaars FG et al (2010) Acquired mutations in the genes encoding IDH1 and IDH2 both are recurrent aberrations in acute myeloid leukemia: prevalence and prognostic value. Blood 116:2122–2126
Rakheja D, Konoplev S, Su M et al (2011) High incidence of IDH mutations in acute myeloid leukaemia with cuplike nuclei. Br J Haematol 155:125–128
Borger DR, Tanabe KK, Fan KC et al (2012) Frequent mutation of isocitrate dehydrogenase (IDH)1 and IDH2 in cholangiocarcinoma identified through broad-based tumor genotyping. Oncologist 17:72–79
Wang P, Dong Q, Zhang C et al (2013) Mutations in isocitrate dehydrogenase 1 and 2 occur frequently in intrahepatic cholangiocarcinomas and share hypermethylation targets with glioblastomas. Oncogene 32:3091–3100
Amary MF, Bacsi K, Maggiani F et al (2011) IDH1 and IDH2 mutations are frequent events in central chondrosarcoma and central and periosteal chondromas but not in other mesenchymal tumours. J Pathol 224:334–343
Gross S, Cairns RA, Minden MD et al (2010) Cancer-associated metabolite 2-hydroxyglutarate accumulates in acute myelogenous leukemia with isocitrate dehydrogenase 1 and 2 mutations. J Exp Med 207:339–344
Dang L, White DW, Gross S et al (2009) Cancer-associated IDH1 mutations produce 2-hydroxyglutarate. Nature 462:739–744
Ward PS, Patel J, Wise DR et al (2010) 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
Choi C, Ganji SK, DeBerardinis RJ et al (2012) 2-hydroxyglutarate detection by magnetic resonance spectroscopy in IDH-mutated patients with gliomas. Nat Med 18:624–629
Moroni I, Bugiani M, D’Incerti L et al (2004) L-2-hydroxyglutaric aciduria and brain malignant tumors: a predisposing condition? Neurology 62:1882–1884
Ozisik PA, Akalan N, Palaoglu S et al (2002) Medulloblastoma in a child with the metabolic disease L-2-hydroxyglutaric aciduria. Pediatr Neurosurg 37:22–26
Haliloglu G, Jobard F, Oguz KK et al (2008) L-2-hydroxyglutaric aciduria and brain tumors in children with mutations in the L2HGDH gene: neuroimaging findings. Neuropediatrics 39:119–122
Shim EH, Livi CB, Rakheja D et al (2014) L-2-Hydroxyglutarate: an epigenetic modifier and putative oncometabolite in renal cancer. Cancer Discov 4:1290–1298
Flavahan WA, Drier Y, Liau BB et al (2016) Insulator dysfunction and oncogene activation in IDH mutant gliomas. Nature 529:10–114
Rakheja D, Medeiros LJ, Bevan S et al (2013) The emerging role of d-2-hydroxyglutarate as an oncometabolite in hematolymphoid and central nervous system neoplasms. Front Oncol 3:169
Figueroa ME, Abdel-Wahab O, Lu C et al (2010) Leukemic IDH1 and IDH2 mutations result in a hypermethylation phenotype, disrupt TET2 function, and impair hematopoietic differentiation. Cancer Cell 18:553–567
Xu W, Yang H, Liu Y et al (2011) Oncometabolite 2-hydroxyglutarate is a competitive inhibitor of alpha-ketoglutarate-dependent dioxygenases. Cancer Cell 19:17–30
Chowdhury R, Yeoh KK, Tian YM et al (2011) The oncometabolite 2-hydroxyglutarate inhibits histone lysine demethylases. EMBO Rep 12:463–469
DiNardo CD, Propert KJ, Loren AW et al (2013) Serum 2-hydroxyglutarate levels predict isocitrate dehydrogenase mutations and clinical outcome in acute myeloid leukemia. Blood 121:4917–4924
Borger DR, Goyal L, Yau T et al (2014) Circulating oncometabolite 2-hydroxyglutarate is a potential surrogate biomarker in patients with isocitrate dehydrogenase-mutant intrahepatic cholangiocarcinoma. Clin Cancer Res 20:1884–1890
Pollyea DA, Kohrt HE, Zhang B et al (2013) 2-Hydroxyglutarate in IDH mutant acute myeloid leukemia: predicting patient responses, minimal residual disease and correlations with methylcytosine and hydroxymethylcytosine levels. Leuk Lymphoma 54:408–410
Natsumeda M, Igarashi H, Nomura T et al (2014) Accumulation of 2-hydroxyglutarate in gliomas correlates with survival: a study by 3.0-tesla magnetic resonance spectroscopy. Acta Neuropathol Commun 2(158):2014
Andronesi OC, Kim GS, Gerstner E et al (2012) Detection of 2-hydroxyglutarate in IDH-mutated glioma patients by in vivo spectral-editing and 2D correlation magnetic resonance spectroscopy. Sci Transl Med 4:116ra114
Janin M, Mylonas E, Saada V et al (2014) Serum 2-hydroxyglutarate production in IDH1- and IDH2-mutated de novo acute myeloid leukemia: a study by the acute leukemia French association group. J Clin Oncol 32:297–305
Sellner L, Capper D, Meyer J et al (2010) Increased levels of 2-hydroxyglutarate in AML patients with IDH1-R132H and IDH2-R140Q mutations. Eur J Haematol 85:457–459
Kalinina J, Carroll A, Wang L et al (2012) Detection of “oncometabolite” 2-hydroxy-glutarate by magnetic resonance analysis as a biomarker of IDH1/2 mutations in glioma. J Mol Med (Berl) 90:1161–1171
McGehee E, Rakheja D, Oliver D et al (2016) The importance of plasma D-2HG measurement in screening for IDH mutations in acute myeloid leukaemia. Br J Haematol 173:323–326
Rakheja D, Boriack RL, Mitui M et al (2011) Papillary thyroid carcinoma shows elevated levels of 2-hydroxyglutarate. Tumour Biol 32:325–323
Rakheja D, Mitui M, Boriack RL et al (2011) Isocitrate dehydrogenase 1/2 mutational analyses and 2-hydroxyglutarate measurements in Wilms tumors. Pediatr Blood Cancer 56:379–383
Struys EA, Jansen EE, Verhoeven NM et al (2004) Measurement of urinary D- and L-2-hydroxyglutarate enantiomers by stable-isotope-dilution liquid chromatography-tandem mass spectrometry after derivatization with diacetyl-L-tartaric anhydride. Clin Chem 50:1391–1395
Struys EA, Verhoeven NM, Roos B et al (2003) Disease-related metabolites in culture medium of fibroblasts from patients with D-2-hydroxyglutaric aciduria, L-2-hydroxyglutaric aciduria, and combined D/L-2-hydroxyglutaric aciduria. Clin Chem 49:1133–1138
Lowry OH, Rosebrough NJ, Farr AL et al (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
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Jones, P.M., Boriack, R., Struys, E.A., Rakheja, D. (2017). Measurement of Oncometabolites d-2-Hydroxyglutaric Acid and l-2-Hydroxyglutaric Acid. In: Fortina, P., Londin, E., Park, J., Kricka, L. (eds) Acute Myeloid Leukemia. Methods in Molecular Biology, vol 1633. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7142-8_14
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DOI: https://doi.org/10.1007/978-1-4939-7142-8_14
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Online ISBN: 978-1-4939-7142-8
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