title: D-2-Hydroxyglutarate producing neo-enzymatic activity inversely correlates with frequency of the type of isocitrate dehydrogenase 1 mutations found in glioma.
abstract: BACKGROUND
  IDH mutations frequently occur in diffuse gliomas and result in a neo-enzymatic activity that results in reduction of α-ketoglutarate to D-2-hydroxyglutarate. In gliomas, the frequency of IDH1 mutations in codon 132 increases in the order R132L-R132S-R132G-R132C-R132H with R132H constituting more than 90% of all IDH1 mutations.
  
  
  RESULTS
  We determined the levels of D-2-hydroxyglutarate in glioma tissues with IDH1 mutations. D-2-hydroxyglutarate levels increased in the order of R132H-R132C-R132S/R132G/R132L. We expressed and purified IDH1 wild type and mutant protein for biochemical characterization. Enzyme kinetics of mutant IDH protein correlated well with D-2-hydroxyglutarate production in cells with R132H exhibiting the highest and R132L the lowest KM for α-ketoglutarate. Addition of D-2-hydroxyglutarate to the medium of cell lines revealed an inhibitory effect at higher concentrations. Migration of LN229 increased at lower D-2-hydroxyglutarate concentrations while higher concentrations showed no effect.
  
  
  CONCLUSIONS
  These findings may suggest natural selection against the rare IDH1R132 mutations in human glioma due to toxicity caused by high levels of D-2-hydroxyglutarate.
authors: Stefan
authors: S
authors: Pusch
authors: Leonille
authors: L
authors: Schweizer
authors: Ann-Christin
authors: AC
authors: Beck
authors: Johanna-Marie
authors: JM
authors: Lehmler
authors: Susanne
authors: S
authors: Weissert
authors: Jörg
authors: J
authors: Balss
authors: Aubry K
authors: AK
authors: Miller
authors: German Consortium of Translational Cancer Research (DKTK), Clinical Cooperation Unit Neuropathology, German Cancer Research Center (DKFZ), INF 280, Heidelberg D-69120, Germany. Andreas.vonDeimling@med.uni-heidelberg.de.
authors: Andreas
authors: A
authors: von Deimling
mesh_terms: D001932:Brain Neoplasms / Q000235:genetics* / Q000378:metabolism; D002465:Cell Movement / Q000235:genetics; D002470:Cell Survival / Q000235:genetics; D005910:Glioma / Q000235:genetics* / Q000378:metabolism; D005977:Glutarates / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D007521:Isocitrate Dehydrogenase / Q000235:genetics* / Q000378:metabolism; D016296:Mutagenesis / Q000235:genetics; D050505:Mutant Proteins; D009154:Mutation / Q000235:genetics*; D014162:Transfection; D014410:Tumor Stem Cell Assay
chemical_list: D005977:Glutarates; D050505:Mutant Proteins; D049452:Green Fluorescent Proteins; C019417:alpha-hydroxyglutarate; D007521:Isocitrate Dehydrogenase; C543588:IDH1 protein, human