Transformation by the (R)-enantiomer of 2-hydroxyglutarate linked to EGLN activation

Abstract

The identification of succinate dehydrogenase (SDH), fumarate hydratase (FH) and isocitrate dehydrogenase (IDH) mutations in human cancers has rekindled the idea that altered cellular metabolism can transform cells. Inactivating SDH and FH mutations cause the accumulation of succinate and fumarate, respectively, which can inhibit 2-oxoglutarate (2-OG)-dependent enzymes, including the EGLN prolyl 4-hydroxylases that mark the hypoxia inducible factor (HIF) transcription factor for polyubiquitylation and proteasomal degradation1. Inappropriate HIF activation is suspected of contributing to the pathogenesis of SDH-defective and FH-defective tumours but can suppress tumour growth in some other contexts. IDH1 and IDH2, which catalyse the interconversion of isocitrate and 2-OG, are frequently mutated in human brain tumours and leukaemias. The resulting mutants have the neomorphic ability to convert 2-OG to the (R)-enantiomer of 2-hydroxyglutarate ((R)-2HG)2,3. Here we show that (R)-2HG, but not (S)-2HG, stimulates EGLN activity, leading to diminished HIF levels, which enhances the proliferation and soft agar growth of human astrocytes. These findings define an enantiomer-specific mechanism by which the (R)-2HG that accumulates in IDH mutant brain tumours promotes transformation and provide a justification for exploring EGLN inhibition as a potential treatment strategy.

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Figure 1: Oncogenic properties of IDH1 R132H.
Figure 2: (R )-2HG can serve as an EGLN cosubstrate.
Figure 3: HIF activity is diminished in IDH mutant cells.
Figure 4: Decreased HIF activity contributes to transformation by mutant IDH.

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Acknowledgements

We thank R. P. Hausinger and J. D. Rabinowitz for helpful suggestions and critical reading of the manuscript, C. Schofield and Y. Zhang for reagents, S. Chen and Y. Shi for JMJD2D assays, K. Koski for modelling and T. Aatsinki and E. Lehtimäki for technical assistance. W.G.K. is a Doris Duke Distinguished Clinical Scholar and a Howard Hughes Medical Institute (HHMI) Investigator. Supported by the National Institutes of Health (W.G.K.), HHMI (W.G.K.), Doris Duke Foundation (W.G.K.), Academy of Finland Grants 120156, 140765 and 218129 (P.K.) and S. Juselius Foundation (P.K.).

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P.K., S.L. and W.G.K. initiated the project, analysed the data and wrote the manuscript. P.K. performed the in vitro enzyme assays. S.L. generated astrocyte cell lines stably expressing various IDH1 proteins. C.G.D., G. Lo. and H.Y. generated the HCT116 subclones. S.R., K.L.L. and S.W. provided oligodendroglioma cell lines. G. Lu generated and validated the reporter plasmids encoding HIF-1α–luciferase fusion proteins. P.J., U.B. and S.G. performed the LC–MS analysis. J.T. synthesized 13C-(R)-2HG and R.L. synthesized and purified different 2-OG and 2-HG derivatives. R.G.W.V. performed the bioinformatics. P.K. and S.L. performed all other experiments with the help of G. Lu, J. A. L. and P.J. All the authors discussed the results and commented on the manuscript.

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Correspondence to William G. Kaelin Jr.

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Koivunen, P., Lee, S., Duncan, C. et al. Transformation by the (R)-enantiomer of 2-hydroxyglutarate linked to EGLN activation. Nature 483, 484–488 (2012). https://doi.org/10.1038/nature10898

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