Isocitrate Dehydrogenase 1/2 Mutational Analyses and 2-Hydroxyglutarate Measurements in Wilms Tumors

Department of Pathology, Children's Medical Center and UT Southwestern Medical Center, Dallas, Texas 75390, USA.
Pediatric Blood & Cancer (Impact Factor: 2.39). 03/2011; 56(3):379-83. DOI: 10.1002/pbc.22697
Source: PubMed


L-2-Hydroxyglutaric aciduria (L-2-HGA) is an uncommon inborn error of metabolism, in which the patients are predisposed to develop brain tumors. Elevated levels of D-2-hydroxyglutarate have been demonstrated with malignant gliomas and myeloid leukemias associated with somatic mutations of the genes encoding NADP(+)-dependent isocitrate dehydrogenases (IDH1 and IDH2, respectively). Recently, we noted a Wilms tumor in a child with L-2-HGA. Given the accumulating evidence that both enantiomers of 2-hydroxyglutarate are associated with cellular transformation, we investigated if sporadic Wilms tumors are associated with IDH1 or IDH2 mutations or with elevated levels of 2-hydroxyglutarate.
We retrieved 21 frozen Wilms tumor tissues. In 20 cases, we sequenced exon 4 and flanking intronic regions of IDH1 and IDH2. In all 21 cases, we measured 2-hydroxyglutarate levels by liquid chromatography-tandem mass spectrometry.
We did not find mutations at the hot spots IDH1 codon 132 or IDH2 codon 172. Two cases (1 with favorable histology and 1 with unfavorable histology) showed heterozygous change c.211G>A (p.Val71Ile) in IDH1, a change previously reported as a mutation but listed as a single nucleotide polymorphism in the NCBI SNP database. We did not find increased levels of 2-hydroxygluatric acid in any sample.
Our results suggest that IDH1 codon 132 or IDH2 codon 172 mutations or elevated 2-hydroxyglutarate levels do not play a role in the biology of sporadic Wilms tumors. The significance of heterozygous change c.211G>A (p.Val71Ile) in IDH1, seen in two tumors, is not clear.

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Available from: Richard L. Boriack, Jun 19, 2015
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    • "At MD Anderson Cancer Center, a Sanger based assay has been recently transitioned to a next generation sequencing assay used to detect IDH1/2 mutations in AML patients at time of initial diagnosis. The metabolites, d-2HG and l-2-hydroxyglutaric acid (l-2HG), are measured using liquid chromatography-tandem mass spectrometry (LC-MS/MS) as described previously (Rakheja et al., 2011a,b,c). "
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    ABSTRACT: Approximately 20% of unselected cases and 30% cytogenetically diploid cases of acute myeloid leukemia (AML) and 80% of grade II-III gliomas and secondary glioblastomas carry mutations in the isocitrate dehydrogenase (IDH) 1 and 2 genes. IDH1/2 mutations prevent oxidative decarboxylation of isocitrate to α-ketoglutarate (α-KG) and modulate the function of IDH (neomorphic activity) thereby facilitating reduction of α-KG to D-2-hydroxyglutarate (D-2HG), a putative oncometabolite. D-2HG is thought to act as a competitive inhibitor of α-KG-dependent dioxygenases that include prolyl hydroxylases and chromatin-modifying enzymes. The end result is a global increase of cellular DNA hypermethylation and alterations of the cellular epigenetic state, which has been proposed to play a role in the development of a variety of tumors. In this review, we provide an update on potential molecular mechanisms linking IDH1/2 mutations and the resulting oncometabolite, D-2HG, with malignant transformation. In addition, in patients with AML and glioma we focus on the associations between IDH1/2 mutations and clinical, morphologic, cytogenetic, and molecular characteristics.
    Frontiers in Oncology 07/2013; 3:169. DOI:10.3389/fonc.2013.00169
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    • "Non-CNS tumors have been documented in L-2-HGA as well, including a bone tumor involving the right frontal region of the calvaria in one patient (Larnaout et al 2007) and nephroblastoma (Wilms tumor) in a second patient (Rogers et al 2010). On the other hand, a subsequent study in 21 Wilms tumor tissues did not reveal increased L-2-HG levels, which therefore did not directly relate L-2-HG with Wilms tumor formation (Rakheja et al 2011). The possible association of the metabolite L-2-HG and tumorigenesis requires further investigation. "
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    ABSTRACT: The organic acidurias D: -2-hydroxyglutaric aciduria (D-2-HGA), L-2-hydroxyglutaric aciduria (L-2-HGA), and combined D,L-2-hydroxyglutaric aciduria (D,L-2-HGA) cause neurological impairment at young age. Accumulation of D-2-hydroxyglutarate (D-2-HG) and/or L-2-hydroxyglutarate (L-2-HG) in body fluids are the biochemical hallmarks of these disorders. The current review describes the knowledge gathered on 2-hydroxyglutaric acidurias (2-HGA), since the description of the first patients in 1980. We report on the clinical, genetic, enzymatic and metabolic characterization of D-2-HGA type I, D-2-HGA type II, L-2-HGA and D,L-2-HGA, whereas for D-2-HGA type I and type II novel clinical information is presented which was derived from questionnaires.
    Journal of Inherited Metabolic Disease 03/2012; 35(4):571-87. DOI:10.1007/s10545-012-9462-5 · 3.37 Impact Factor
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    • "Additional IDH mutants distinguished by R(À)-2HG production PS Ward et al tically characterized (Table 1). We first examined the effects of reported IDH1 SNPs, V71I and V178I, as well as non-arginine variants only reported in a single sample to date (Hemerly et al., 2010; Ho et al., 2010; Marcucci et al., 2010; Murugan et al., 2010; Zou et al., 2010; Forbes et al., 2011; Rakheja et al., 2011). All were expressed at comparable levels to IDH1/2 WT in cells (Supplementary Figure 1). "
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    ABSTRACT: Mutations in cytosolic isocitrate dehydrogenase 1 (IDH1) or its mitochondrial homolog IDH2 can lead to R(-)-2-hydroxyglutarate (2HG) production. To date, mutations in three active site arginine residues, IDH1 R132, IDH2 R172 and IDH2 R140, have been shown to result in the neomorphic production of 2HG. Here we report on three additional 2HG-producing IDH1 mutations: IDH1 R100, which is affected in adult glioma, IDH1 G97, which is mutated in colon cancer cell lines and pediatric glioblastoma, and IDH1 Y139. All these new mutants stereospecifically produced 2HG's (R) enantiomer. In contrast, we find that the IDH1 SNPs V71I and V178I, as well as a number of other single-sample reports of IDH non-synonymous mutation, did not elevate cellular 2HG levels in cells and retained the wild-type ability for isocitrate-dependent NADPH production. Finally, we report the existence of additional rare, but recurring mutations found in lymphoma and thyroid cancer, which while failing to elevate 2HG nonetheless displayed loss of function, indicating a possible tumorigenic mechanism for a non-2HG-producing subset of IDH mutations in some malignancies. These data broaden our understanding of how IDH mutations may contribute to cancer through either neomorphic R(-)-2HG production or reduced wild-type enzymatic activity, and highlight the potential value of metabolite screening in identifying IDH-mutated tumors associated with elevated oncometabolite levels.
    Oncogene 09/2011; 31(19):2491-8. DOI:10.1038/onc.2011.416 · 8.46 Impact Factor
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