IDH1(R132H) mutation increases murine haematopoietic progenitors and alters epigenetics

Abstract

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.

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Figure 1: LysM-KI mice show age-dependent splenomegaly and decreased bone marrow cellularity.
Figure 2: LysM-KI mice show age-dependent increases in lineage-restricted progenitors and extramedullary haematopoiesis.
Figure 3: Altered methylation of DNA and histones in LysM-KI cells.

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Primary accessions

Gene Expression Omnibus

Data deposits

The microarray and sequencing data have been deposited in the Gene ExpressionOmnibus of the National Center for Biotechnical Information with the accession numbers GSE38589 and GSE38687.

Change history

  • 29 August 2012

    The spelling of an author name (S.M.S.) was corrected

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Acknowledgements

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.

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Contributions

C.B.K., M.S. and T.W.M. initiated the project. M.S. designed the conditional IDH1(R132H)-KI mouse and generated it, as well as the IDH1(R132H)-KI embryonic stem cell clones with the help of S.S., S.M.S., A.W., J.H. and A.Y.-T.; C.B.K. designed and organized experiments involving the hematopoietic system in consultation with T.W.M., M.E.F., A.M., J.-C.Z.-P., G.R., P.S.O., C.V. and D.L.B.; C.B.K. performed the flow cytometry analyses, bone marrow transfers, cell sorting, RNA extraction and real-time PCR analyses, immunoblotting, colony formation assays and serial plating experiments. J.C.M. performed the LC-MS analyses. E.F.L., D.B., A.B. and I.S.H. helped with flow cytometric analyses and BM transfers. R.H. performed the OP9/OP9-DL1 embryonic stem cell in vitro differentiation experiments. W.Y.L. helped with flow cytometric analyses and cell culture experiments. M.E.F. and A.M. performed the DNA methylation experiments and analysed the data. C.V. performed the mRNA expression microarray experiments and analysed data. C.B.K. and T.W.M. wrote the manuscript with the help of A.B. and D.B. All authors discussed the results extensively and agree with the conclusions presented in the manuscript.

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Correspondence to Tak W. Mak.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Information

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)

Supplementary Table 3

This table contains differentially expressed mRNAs in LysM-KI and control LSK cells - see Supplementary Information file for full legend. (XLS 77 kb)

Supplementary Table 4

This table contains Gene Ontology categories of the differentially expressed mRNAs - see Supplementary Information file for full legend. (XLS 836 kb)

Supplementary Table 5

This table contains differentially methylated genomic regions - see Supplementary Information file for full legend. (XLS 219 kb)

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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

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