title: Functions of idh1 and its mutation in the regulation of developmental hematopoiesis in zebrafish.
abstract: Isocitrate dehydrogenase 1 mutation (IDH1-R132H) was recently identified in acute myeloid leukemia with normal cytogenetics. The mutant enzyme is thought to convert α-ketoglutarate to the pathogenic 2-hydroxyglutarate (2-HG) that affects DNA methylation via inhibition of ten-eleven translocation 2. However, the role of wild-type IDH1 in normal hematopoiesis and its relevance to acute myeloid leukemia is unknown. Here we showed that zebrafish idh1 (zidh1) knockdown by morpholino and targeted mutagenesis by transcription activator-like effector nuclease might induce blockade in myeloid differentiation, as evident by an increase in pu.1 and decrease in mpo, l-plastin, and mpeg1 expression, and significantly reduce definitive hematopoiesis. Morpholino knockdown of zidh2 also induced a blockade in myeloid differentiation but definitive hematopoiesis was not affected. The hematopoietic phenotype of zidh1 knockdown was not rescuable by zidh2 messenger RNA, suggesting nonredundant functions. Overexpression of human IDH1-R132H or its zebrafish ortholog resulted in 2-HG elevation and expansion of myelopoiesis in zebrafish embryos. A human IDH1-R132H-specific inhibitor (AGI-5198) significantly ameliorated both hematopoietic and 2-HG responses in human but not zebrafish IDH1 mutant expression. The results provided important insights to the role of zidh1 in myelopoiesis and definitive hematopoiesis and of IDH1-R132H in leukemogenesis.
authors.affiliation: Division of Haematology, Department of Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong, China;
authors.forename: Xiangguo
authors.initials: X
authors.lastname: Shi
authors.affiliation: Division of Haematology, Department of Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong, China;
authors.forename: Bai-Liang
authors.initials: BL
authors.lastname: He
authors.affiliation: Division of Haematology, Department of Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong, China;
authors.forename: Alvin C H
authors.initials: AC
authors.lastname: Ma
authors.affiliation: Division of Haematology, Department of Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong, China;
authors.forename: Yuhan
authors.initials: Y
authors.lastname: Guo
authors.affiliation: Department of Cell Biology, Southern Medical University, Guangzhou, China;
authors.forename: Yali
authors.initials: Y
authors.lastname: Chi
authors.affiliation: Division of Haematology, Department of Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong, China;
authors.forename: Cheuk Him
authors.initials: CH
authors.lastname: Man
authors.affiliation: Department of Cell Biology, Southern Medical University, Guangzhou, China;
authors.forename: Wenqing
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Department of Cell Biology, Southern Medical University, Guangzhou, China;
authors.forename: Yiyue
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Molecular Neuroscience, Division of Life Science, Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong, China;
authors.forename: Zilong
authors.initials: Z
authors.lastname: Wen
authors.affiliation: State Key Laboratory of Experimental Hematology, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin, China; and Center for Stem Cell Medicine, Chinese Academy of Medical Sciences, Beijing, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Cheng
authors.affiliation: Division of Haematology, Department of Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong, China;
authors.forename: Anskar Y H
authors.initials: AY
authors.lastname: Leung
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D044482:Benzeneacetamides / Q000494:pharmacology; D015153:Blotting, Western; D002478:Cells, Cultured; D004625:Embryo, Nonmammalian / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D008401:Gas Chromatography-Mass Spectrometry; D005977:Glutarates / Q000378:metabolism; D006410:Hematopoiesis / Q000502:physiology*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007093:Imidazoles / Q000494:pharmacology; D007124:Immunoenzyme Techniques; D007521:Isocitrate Dehydrogenase / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D009154:Mutation / Q000235:genetics*; D038042:Myelopoiesis / Q000502:physiology*; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015027:Zebrafish / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism
chemical_list: C581156:AGI-5198; D044482:Benzeneacetamides; D005977:Glutarates; D007093:Imidazoles; D012333:RNA, Messenger; C019417:alpha-hydroxyglutarate; D007521:Isocitrate Dehydrogenase