Functions of idh1 and its mutation in the regulation of developmental hematopoiesis in zebrafish

Blood. 2015 May 7;125(19):2974-84. doi: 10.1182/blood-2014-09-601187. Epub 2015 Mar 16.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Benzeneacetamides / pharmacology
  • Blotting, Western
  • Cells, Cultured
  • Embryo, Nonmammalian / cytology
  • Embryo, Nonmammalian / drug effects
  • Embryo, Nonmammalian / metabolism*
  • Enzyme-Linked Immunosorbent Assay
  • Flow Cytometry
  • Gas Chromatography-Mass Spectrometry
  • Glutarates / metabolism
  • Hematopoiesis / physiology*
  • Humans
  • Image Processing, Computer-Assisted
  • Imidazoles / pharmacology
  • Immunoenzyme Techniques
  • Isocitrate Dehydrogenase / antagonists & inhibitors
  • Isocitrate Dehydrogenase / genetics*
  • Isocitrate Dehydrogenase / metabolism*
  • Mutagenesis, Site-Directed
  • Mutation / genetics*
  • Myelopoiesis / physiology*
  • RNA, Messenger / genetics
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcriptase Polymerase Chain Reaction
  • Zebrafish / genetics
  • Zebrafish / growth & development*
  • Zebrafish / metabolism

Substances

  • AGI-5198
  • Benzeneacetamides
  • Glutarates
  • Imidazoles
  • RNA, Messenger
  • alpha-hydroxyglutarate
  • Isocitrate Dehydrogenase