Mutations in the IDH1 and IDH2 genes encoding isocitrate dehydrogenases are frequently found in human glioblastomas1 and cytogenetically normal acute myeloid leukaemias (AML)2. These alterations are gain-of-function mutations in that they drive the synthesis of the ‘oncometabolite’ R-2-hydroxyglutarate (2HG)3. It remains unclear how IDH1 and IDH2 mutations modify myeloid cell development and promote leukaemogenesis. Here we report the characterization of conditional knock-in (KI) mice in which the most common IDH1 mutation, IDH1(R132H), is inserted into the endogenous murine Idh1 locus and is expressed in all haematopoietic cells (Vav-KI mice) or specifically in cells of the myeloid lineage (LysM-KI mice). These mutants show increased numbers of early haematopoietic progenitors and develop splenomegaly and anaemia with extramedullary haematopoiesis, suggesting a dysfunctional bone marrow niche. Furthermore, LysM-KI cells have hypermethylated histones and changes to DNA methylation similar to those observed in human IDH1- or IDH2-mutant AML. To our knowledge, our study is the first to describe the generation and characterization of conditional IDH1(R132H)-KI mice, and also the first report to demonstrate the induction of a leukaemic DNA methylation signature in a mouse model. Our report thus sheds light on the mechanistic links between IDH1 mutation and human AML.
Subscribe to Journal
Get full journal access for 1 year
only $3.90 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Parsons, D. W. et al. An integrated genomic analysis of human glioblastoma multiforme. Science 321, 1807–1812 (2008)
Mardis, E. R. et al. Recurring mutations found by sequencing an acute myeloid leukemia genome. N. Engl. J. Med. 361, 1058–1066 (2009)
Dang, L. et al. Cancer-associated IDH1 mutations produce 2-hydroxyglutarate. Nature 462, 739–744 (2009)
Xu, W. et al. Oncometabolite 2-hydroxyglutarate is a competitive inhibitor of α-ketoglutarate-dependent dioxygenases. Cancer Cell 19, 17–30 (2011)
Figueroa, M. E. et al. Leukemic IDH1 and IDH2 mutations result in a hypermethylation phenotype, disrupt TET2 function, and impair hematopoietic differentiation. Cancer Cell 18, 553–567 (2010)
Chowdhury, R. et al. The oncometabolite 2-hydroxyglutarate inhibits histone lysine demethylases. EMBO Rep. 12, 463–469 (2011)
Clausen, B. E. et al. Conditional gene targeting in macrophages and granulocytes using LysMcre mice. Transgenic Res. 8, 265–277 (1999)
Ye, M. et al. Hematopoietic stem cells expressing the myeloid lysozyme gene retain long-term, multilineage repopulation potential. Immunity 19, 689–699 (2003)
Goardon, N. et al. Coexistence of LMPP-like and GMP-like leukemia stem cells in acute myeloid leukemia. Cancer Cell 19, 138–152 (2011)
Holmes R & Zúñiga-Pflücker J.-C The OP9–DL1 system: generation of T-lymphocytes from embryonic or hematopoietic stem cells in vitro. Cold Spring Harb. Protoc. http://dx.doi.org/10.1101/pdb.prot5156 (2009)
de Boer, J. et al. Transgenic mice with hematopoietic and lymphoid specific expression of Cre. Eur. J. Immunol. 33, 314–325 (2003)
Ito, K. et al. Reactive oxygen species act through p38 MAPK to limit the lifespan of hematopoietic stem cells. Nature Med. 12, 446–451 (2006)
Tothova, Z. et al. FoxOs are critical mediators of hematopoietic stem cell resistance to physiologic oxidative stress. Cell 128, 325–339 (2007)
Callens, C. et al. Targeting iron homeostasis induces cellular differentiation and synergizes with differentiating agents in acute myeloid leukemia. J. Exp. Med. 207, 731–750 (2010)
Eliasson, P. & Jönsson, J. I. The hematopoietic stem cell niche: low in oxygen but a nice place to be. J. Cell. Physiol. 222, 17–22 (2010)
Zhao, S. et al. Glioma-derived mutations in IDH1 dominantly inhibit IDH1 catalytic activity and induce HIF-1α. Science 324, 261–265 (2009)
Noushmehr, H. et al. Identification of a CpG island methylator phenotype that defines a distinct subgroup of glioma. Cancer Cell 17, 510–522 (2010)
Akalin, A. et al. Base-pair resolution DNA methylation sequencing reveals profoundly divergent epigenetic landscapes in acute myeloid leukemia. PLoS Genet. (in the press)
Campbell, C. et al. Signal control of hematopoietic stem cell fate: Wnt, Notch, and Hedgehog as the usual suspects. Curr. Opin. Hematol. 15, 319–325 (2008)
Heidel, F. H. et al. Self-renewal related signaling in myeloid leukemia stem cells. Int. J. Hematol. 94, 109–117 (2011)
Klinakis, A. et al. A novel tumour-suppressor function for the Notch pathway in myeloid leukaemia. Nature 473, 230–233 (2011)
Söderberg, S. S. et al. Complex and context dependent regulation of hematopoiesis by TGF-β superfamily signaling. Ann. NY Acad. Sci. 1176, 55–69 (2009)
Patel, K. P. et al. Acute myeloid leukemia with IDH1 or IDH2 mutation: frequency and clinicopathologic features. Am. J. Clin. Pathol. 135, 35–45 (2011)
Patnaik, M. M. et al. Differential prognostic effect of IDH1 versus IDH2 mutations in myelodysplastic syndromes: a Mayo Clinic Study of 277 patients. Leukemia 26, 101–105 (2012)
Munger, J. et al. Systems-level metabolic flux profiling identifies fatty acid synthesis as a target for antiviral therapy. Nature Biotechnol. 26, 1179–1186 (2008)
Bajad, S. U. et al. Separation and quantitation of water soluble cellular metabolites by hydrophilic interaction chromatography-tandem mass spectrometry. J. Chromatogr. A 1125, 76–88 (2006)
Meissner, A. et al. Reduced representation bisulfite sequencing for comparative high-resolution DNA methylation analysis. Nucleic Acids Res. 33, 5868–5877 (2005)
Krueger, F. & Andrews, S. R. Bismark: a flexible aligner and methylation caller for Bisulfite-Seq applications. Bioinformatics 27, 1571–1572 (2011)
Fujita, P. A. et al. The UCSC Genome Browser database: update 2011. Nucleic Acids Res. 39, D876–D882 (2011)
Storey, J. D. & Tibshirani, R. Statistical significance for genomewide studies. Proc. Natl Acad. Sci. USA 100, 9440–9445 (2003)
Du, P. et al. lumi: a pipeline for processing Illumina microarray. Bioinformatics 24, 1547–1548 (2008)
Benjamini, Y. & Hochberg, Y. Controlling the false discovery rate: a practical and powerful approach to multiple testing. J. R. Stat. Soc., B 57, 289–300 (1995)
Smyth G. K Linear models and empirical Bayes methods for assessing differential expression in microarray experiments. Stat. Appl. Genet. Mol. Biol. 3, 3 (2004)
Benjamini, Y. & Yekutieli, D. The control of the false discovery rate in multiple testing under dependency. Ann. Stat. 29, 1165–1188 (2001)
We thank the Animal Research Colony (ARC) at the Ontario Cancer Institute for mouse care; I. Ng, A. Shahinian, J. Sylvester and S. McCracken for administrative and organizational expertise; M. Bailey and J. Tsao for technical assistance; F. Tong and R. Nayyar for assistance with flow cytometric analysis and sorting; the Weill Cornell Medical College (WCMC) Epigenomics Core Facility for technical help and expertise; G. Melino, D. Green, M. Minden, H. Chang and P. Lang for helpful discussions; J. Thomsen for figure layout and M. Saunders for scientific editing. C.B.K. and D.B. were supported in part by a Feodor-Lynen Postdoctoral Research Fellowship from the Alexander-von-Humboldt-Foundation, Germany. D.B. and A.B. were supported in part by a Fellowship from the German Research Foundation (DFG). J.C.M. is supported by a National Institute of Health grant (NIH R01AI081773) and is a Damon Runyon-Rachleff Innovation Awardee supported by the Damon Runyon Cancer Research Foundation (DRR-09-10). P.S.O. holds a Canada Research Chair in Autoimmunity and Tumor Immunity. M.E.F. is supported by the Leukemia & Lymphoma Society Special Fellow Award and a Doris Duke Clinical Scientist Development Award. A.M. is supported by an LLS SCOR grant (7132-08), a Burroughs Wellcome Clinical Translational Scientist Award and a Starr Cancer Consortium grant (I4-A442). J.-C.Z.-P. is supported by a Canada Research Chair in Developmental Immunology. This work was supported by grants from the Canadian Institutes of Health Research (CIHR) and the Ontario Ministry of Health and Long Term Care to T.W.M., and a program grant from the Terry Fox Foundation to P.S.O., J.-C.Z.-P. and T.W.M. Please note that the views expressed do not necessarily reflect those of the OMOHLTC.
The authors declare no competing financial interests.
This file contains Supplementary Figures 1-11, Supplementary Tables 1-2 and 6-7 and legends for Supplementary Tables 3, 4 and 5 (see separate files for these tables). (PDF 1609 kb)
This table contains differentially expressed mRNAs in LysM-KI and control LSK cells - see Supplementary Information file for full legend. (XLS 77 kb)
This table contains Gene Ontology categories of the differentially expressed mRNAs - see Supplementary Information file for full legend. (XLS 836 kb)
This table contains differentially methylated genomic regions - see Supplementary Information file for full legend. (XLS 219 kb)
About this article
Cite this article
Sasaki, M., Knobbe, C., Munger, J. et al. IDH1(R132H) mutation increases murine haematopoietic progenitors and alters epigenetics. Nature 488, 656–659 (2012). https://doi.org/10.1038/nature11323