title: Fumarate and Succinate Regulate Expression of Hypoxia-inducible Genes via TET Enzymes.
abstract: The TET enzymes are members of the 2-oxoglutarate-dependent dioxygenase family and comprise three isoenzymes in humans: TETs 1-3. These TETs convert 5-methylcytosine to 5-hydroxymethylcytosine (5-hmC) in DNA, and high 5-hmC levels are associated with active transcription. The importance of the balance in these modified cytosines is emphasized by the fact that TET2 is mutated in several human cancers, including myeloid malignancies such as acute myeloid leukemia (AML). We characterize here the kinetic and inhibitory properties of Tets and show that the Km value of Tets 1 and 2 for O2 is 30 μm, indicating that they retain high activity even under hypoxic conditions. The AML-associated mutations in the Fe(2+) and 2-oxoglutarate-binding residues increased the Km values for these factors 30-80-fold and reduced the Vmax values. Fumarate and succinate, which can accumulate to millimolar levels in succinate dehydrogenase and fumarate hydratase-mutant tumors, were identified as potent Tet inhibitors in vitro, with IC50 values ∼400-500 μm. Fumarate and succinate also down-regulated global 5-hmC levels in neuroblastoma cells and the expression levels of some hypoxia-inducible factor (HIF) target genes via TET inhibition, despite simultaneous HIFα stabilization. The combination of fumarate or succinate treatment with TET1 or TET3 silencing caused differential effects on the expression of specific HIF target genes. Altogether these data show that hypoxia-inducible genes are regulated in a multilayered manner that includes epigenetic regulation via TETs and 5-hmC levels in addition to HIF stabilization.
authors.affiliation: From the Biocenter Oulu, Faculty of Biochemistry and Molecular Medicine, Oulu Center for Cell-Matrix Research, University of Oulu, FIN-90014 Oulu, Finland.
authors.forename: Tuomas
authors.initials: T
authors.lastname: Laukka
authors.affiliation: the Department of Medicine, Section of Hematology/Oncology, the Committee on Molecular Pathogenesis and Molecular Medicine and.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Mariani
authors.affiliation: From the Biocenter Oulu, Faculty of Biochemistry and Molecular Medicine, Oulu Center for Cell-Matrix Research, University of Oulu, FIN-90014 Oulu, Finland.
authors.forename: Tuukka
authors.initials: T
authors.lastname: Ihantola
authors.affiliation: the Department of Medicine, Section of Hematology/Oncology, the Committee on Cancer Biology, University of Chicago, Chicago, Illinois 60637.
authors.forename: John Z
authors.initials: JZ
authors.lastname: Cao
authors.affiliation: Admescope Ltd., Typpitie 1, FIN-90620 Oulu, Finland.
authors.forename: Juho
authors.initials: J
authors.lastname: Hokkanen
authors.affiliation: the Department of Medical Oncology, Dana-Farber Cancer Institute and Brigham and Women's Hospital, Boston, Massachusetts 02215, and the Howard Hughes Medical Institute, Chevy Chase, Maryland 20815.
authors.forename: William G
authors.initials: WG
authors.lastname: Kaelin
authors.affiliation: the Department of Medicine, Section of Hematology/Oncology, the Committee on Cancer Biology, University of Chicago, Chicago, Illinois 60637.
authors.forename: Lucy A
authors.initials: LA
authors.lastname: Godley
authors.affiliation: From the Biocenter Oulu, Faculty of Biochemistry and Molecular Medicine, Oulu Center for Cell-Matrix Research, University of Oulu, FIN-90014 Oulu, Finland, peppi.koivunen@oulu.fi.
authors.forename: Peppi
authors.initials: P
authors.lastname: Koivunen
mesh_terms: D000818:Animals; D015687:Cell Hypoxia; D045744:Cell Line, Tumor; D004268:DNA-Binding Proteins / Q000096:biosynthesis* / Q000235:genetics; D049308:Dioxygenases / Q000096:biosynthesis* / Q000235:genetics; D005650:Fumarates / Q000494:pharmacology*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D051379:Mice; D006899:Mixed Function Oxygenases; D009154:Mutation; D009447:Neuroblastoma / Q000235:genetics / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000096:biosynthesis* / Q000235:genetics; D019802:Succinic Acid / Q000494:pharmacology*
chemical_list: D004268:DNA-Binding Proteins; D005650:Fumarates; D011518:Proto-Oncogene Proteins; C032005:fumaric acid; D019802:Succinic Acid; D006899:Mixed Function Oxygenases; C462539:TET1 protein, human; C578792:TET3 protein, human; D049308:Dioxygenases
keywords: 5-hydroxymethylcytosine (5-hmC); AML; TET; epigenetics; fumarate; hypoxia; hypoxia-inducible factor (HIF); leukemia; succinate