Lactate Dehydrogenase C Produces S-2-Hydroxyglutarate in Mouse Testis

Teng, Xin; Emmett, Matthew J.; Lazar, Mitchell A.; Goldberg, Erwin; Rabinowitz, Joshua D.

doi:10.1021/acschembio.6b00290

Cited by 40 publications

(46 citation statements)

References 40 publications

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“…S5A). In agreement with earlier studies (16,21), we also observed that human LDHA catalyzed the reaction of 2OG to L-2HG, thereby demonstrating that LDH generates L-2HG via an evolutionarily conserved enzymatic mechanism ( Fig. 3B and Fig.…”

Section: Resultssupporting

confidence: 92%

“…2 D and E). Furthermore, these metabolic defects were specifically due to the loss of dLDH activity, as the aberrant pyruvate, lactate, and 2HG levels observed in dLdh 16 /dLdh 17 mutants were completely rescued by a dLdh transgene (Fig. 2 D and E).…”

Section: Resultsmentioning

confidence: 95%

“…S3 C and D). However, despite the normal growth rate, GC-MS analysis of the dLdh 16 /dLdh 17 midsecond-instar larvae revealed dramatic metabolic phenotypes. These mutants not only displayed a fivefold increase in pyruvate and a 20-fold decrease in lactate but 2HG levels were almost undetectable compared with a precise excision control strain (dLdh prec ) ( Fig.…”

Section: Resultsmentioning

confidence: 98%

“…S3 A and B). Animals that harbor a trans-heterozygous combination of these null alleles, dLdh 16 /dLdh 17 , grow at a normal rate until the midthird instar, at which point 50% of the larvae die and the remainder develop into morphologically normal adults ( Fig. S3 C and D).…”

Section: Resultsmentioning

confidence: 99%

“…Unlike D-2HG, however, L-2HG has no known role in metabolism and eukaryotes lack an enzyme dedicated to L-2HG synthesis. Instead, L-2HG is considered an aberrant metabolite produced by the nonspecific activity of enzymes such as malate dehydrogenase, lactate dehydrogenase A (LDHA), and lactate dehydrogenase C (12)(13)(14)(15)(16)(17)(18). Consistent with these observations, L-2HG accumulation in cancer cells does not result from ectopic synthesis; rather, it stems from decreased expression of the enzyme L-2-hydroxyglutarate dehydrogenase (L2HGDH), which is solely responsible for L-2HG degradation (10,19).…”

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confidence: 99%

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Drosophila larvae synthesize the putative oncometabolite L-2-hydroxyglutarate during normal developmental growth

Chawla

Hurlburt

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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L-2-hydroxyglutarate (L-2HG) has emerged as a putative oncometabolite that is capable of inhibiting enzymes involved in metabolism, chromatin modification, and cell differentiation. However, despite the ability of L-2HG to interfere with a broad range of cellular processes, this molecule is often characterized as a metabolic waste product. Here, we demonstrate that Drosophila larvae use the metabolic conditions established by aerobic glycolysis to both synthesize and accumulate high concentrations of L-2HG during normal developmental growth. A majority of the larval L-2HG pool is derived from glucose and dependent on the Drosophila estrogen-related receptor (dERR), which promotes L-2HG synthesis by up-regulating expression of the Drosophila homolog of lactate dehydrogenase (dLdh). We also show that dLDH is both necessary and sufficient for directly synthesizing L-2HG and the Drosophila homolog of L-2-hydroxyglutarate dehydrogenase (dL2HGDH), which encodes the enzyme that breaks down L-2HG, is required for stage-specific degradation of the L-2HG pool. In addition, dLDH also indirectly promotes L-2HG accumulation via synthesis of lactate, which activates a metabolic feedforward mechanism that inhibits dL2HGDH activity and stabilizes L-2HG levels. Finally, we use a genetic approach to demonstrate that dLDH and L-2HG influence position effect variegation and DNA methylation, suggesting that this compound serves to coordinate glycolytic flux with epigenetic modifications. Overall, our studies demonstrate that growing animal tissues synthesize L-2HG in a controlled manner, reveal a mechanism that coordinates glucose catabolism with L-2HG synthesis, and establish the fly as a unique model system for studying the endogenous functions of L-2HG during cell growth and proliferation.ne of the hallmarks of cancer is a dramatic reprograming of cellular metabolism that results in enhanced biosynthesis (1). These metabolic changes are particularly apparent in tumors that use the Warburg effect, also referred to as aerobic glycolysis, a metabolic program characterized by elevated levels of glucose consumption and enhanced lactate production (1, 2). By activating aerobic glycolysis, tumors are able to synthesize macromolecules rapidly from glycolytic intermediates. In addition, elevated levels of lactate dehydrogenase (LDH) activity allow proliferating cells to synthesize lactate and maintain the NAD + levels required for high rates of glucose catabolism and biomass production (1).The metabolic reprogramming of cancer cells, however, extends beyond biosynthesis, as many tumors also generate progrowth metabolites, or oncometabolites, that promote tumor formation via nonmetabolic means. Most notable among these compounds is D-2-hydroxyglutarate (D-2HG), which is associated with cancers such as gliomas and acute myelogenous leukemias (3). Although D-2HG is generated as a normal byproduct of γ-hydroxybutyrate metabolism (4), oncogenic D-2HG production is the result of neomorphic mutations in the active site of isocitrate dehydrogenas...

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Section: Resultssupporting

confidence: 92%

Section: Resultsmentioning

confidence: 95%

Section: Resultsmentioning

confidence: 98%

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

Drosophila larvae synthesize the putative oncometabolite L-2-hydroxyglutarate during normal developmental growth

Chawla

Hurlburt

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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Separation and determination of the enantiomers of lactic acid and 2‐hydroxyglutaric acid by chiral derivatization combined with gas chromatography and mass spectrometry

Ding

Lin

Weng

et al. 2018

J of Separation Science

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Lactic acid and 2-hydroxyglutaric acid are chiral metabolites that have two distinct d- and l-enantiomers with distinct biochemical properties. Perturbations of a single enantiomeric form have been found to be closely related to certain diseases. Therefore, the ability to differentiate the d and l enantiomers is important for these disease studies. Herein, we describe a method for the separation and determination of lactic acid and 2-hydroxyglutaric acid enantiomers by chiral derivatization (with l-menthol and acetyl chloride) combined with gas chromatography and mass spectrometry. The two pairs of above-mentioned enantiomers exhibited linear calibration curves with a correlation coefficient (R ) exceeding 0.99. The measured data were accurate in the acceptable recovery range of 88.17-102.30% with inter- and intraday precisions (relative standard deviations) in the range of 4.23-17.26%. The limits of detection for d-lactic acid, l-lactic acid, d-2-hydroxyglutaric acid, and l-2-hydroxyglutaric acid were 0.13, 0.11, 1.12, and 1.16 μM, respectively. This method was successfully applied to analyze mouse plasma. The d-lactic acid levels in type 2 diabetes mellitus mouse plasma were observed to be significantly higher (P < 0.05, t-test) than those of normal mice, suggesting that d-lactic acid may serve as an indicator for type 2 diabetes mellitus.

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Synthesis of [1‐¹³C‐5‐¹²C]‐alpha‐ketoglutarate enables noninvasive detection of 2‐hydroxyglutarate

et al. 2021

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Isocitrate dehydrogenase 1 (IDH1) mutations that generate the oncometabolite 2-hydroxyglutarate (2-HG) from α-ketoglutarate (α-KG) have been identified in many types of tumors and are an important prognostic factor in gliomas. 2-HG production can be determined by hyperpolarized carbon-13 magnetic resonance spectroscopy (HP-13 C-MRS) using [1-13 C]-α-KG as a probe, but peak contamination from naturally occurring [5-13 C]-α-KG overlaps with the [1-13 C]-2-HG peak. Via a newly developed oxidative-Stetter reaction, [1-13 C-5-12 C]-α-KG was synthesized. α-KG metabolism was measured via HP-13 C-MRS using [1-13 C-5-12 C]-α-KG as a probe. [1-13 C-5-12 C]α-KG was synthesized in high yields, and successfully eliminated the signal from C5 of α-KG in the HP-13 C-MRS spectra. In HCT116 IDH1 R132H cells, [1-13 C-5-12 C]α-KG allowed for unimpeded detection of [1-13 C]-2-HG. 12 C-enrichment represents a novel method to circumvent spectral overlap, and [1-13 C-5-12 C]-α-KG shows promise as a probe to study IDH1 mutant tumors and α-KG metabolism.2-hydroxyglutarate, alpha-ketoglutarate, carbon-13 labeled metabolites, hyperpolarized carbon-13 nuclear magnetic resonance spectroscopy, isocitrate dehydrogenase 1 | INTRODUCTIONAtypical metabolism of alpha-ketoglutarate (α-KG) has been linked to increased tumor cell differentiation 1 and amplified malignant progression 1 as well as altered protein synthesis and catabolism. 2 α-KG is also the substrate for the mutant isocitrate dehydrogenase 1 (IDH1) enzyme. IDH1 is a cytosolic enzyme that catalyzes the oxidation of isocitrate to α-KG. Mutations in this enzyme, most commonly a heterozygous point mutation of arginine 132 to histidine (R132H), instead allow for the reduction of α-KG to 2-hydroxyglutarate (2-HG). 3 This gain of function mutation leads to a buildup of 2-HG in IDH1 mutant cells. 3 As pyruvate and α-KG are structurally similar, 2-HG can also be produced through noncanonical activity of lactate dehydrogenase (LDH), but these promiscuous reactions by LDH do not typically lead to high concentrations of 2-HG in human Abbreviations used: 2-HG, 2-hydroxyglutarate; HP, hyperpolarized; IDH1, isocitrate dehydrogenase 1; LDH, lactate dehydrogenase; MRS, magnetic resonance spectroscopy; NADH, reduced nicotinamide adenine dinucleotide; α-KG, alpha-ketoglutarate. Natsuko Miura and Chandrasekhar Mushti contributed equally to this work.

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Lactate Dehydrogenase C Produces S-2-Hydroxyglutarate in Mouse Testis

Cited by 40 publications

References 40 publications

Drosophila larvae synthesize the putative oncometabolite L-2-hydroxyglutarate during normal developmental growth

Drosophila larvae synthesize the putative oncometabolite L-2-hydroxyglutarate during normal developmental growth

Separation and determination of the enantiomers of lactic acid and 2‐hydroxyglutaric acid by chiral derivatization combined with gas chromatography and mass spectrometry

Synthesis of [1‐¹³C‐5‐¹²C]‐alpha‐ketoglutarate enables noninvasive detection of 2‐hydroxyglutarate

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Lactate Dehydrogenase C Produces S-2-Hydroxyglutarate in Mouse Testis

Cited by 40 publications

References 40 publications

Drosophila larvae synthesize the putative oncometabolite L-2-hydroxyglutarate during normal developmental growth

Drosophila larvae synthesize the putative oncometabolite L-2-hydroxyglutarate during normal developmental growth

Separation and determination of the enantiomers of lactic acid and 2‐hydroxyglutaric acid by chiral derivatization combined with gas chromatography and mass spectrometry

Synthesis of [1‐13C‐5‐12C]‐alpha‐ketoglutarate enables noninvasive detection of 2‐hydroxyglutarate

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Synthesis of [1‐¹³C‐5‐¹²C]‐alpha‐ketoglutarate enables noninvasive detection of 2‐hydroxyglutarate