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Early Lineage Priming by Trisomy of Erg Leads to Myeloproliferation in a Down Syndrome Model

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Early Lineage Priming by Trisomy of Erg Leads to Myeloproliferation in a Down Syndrome Model Early Lineage Priming by Trisomy of Erg Leads to Myeloproliferation in a Down Syndrome Model The Harvard community has made this article openly available. Please share how this access benefits you. Your story matters Citation Ng, A. P., Y. Hu, D. Metcalf, C. D. Hyland, H. Ierino, B. Phipson, D. Wu, et al. 2015. “Early Lineage Priming by Trisomy of Erg Leads to Myeloproliferation in a Down Syndrome Model.” PLoS Genetics 11 (5): e1005211. doi:10.1371/journal.pgen.1005211. http:// dx.doi.org/10.1371/journal.pgen.1005211. Published Version doi:10.1371/journal.pgen.1005211 Citable link http://nrs.harvard.edu/urn-3:HUL.InstRepos:16120953 Terms of Use This article was downloaded from Harvard University’s DASH repository, and is made available under the terms and conditions applicable to Other Posted Material, as set forth at http:// nrs.harvard.edu/urn-3:HUL.InstRepos:dash.current.terms-of- use#LAA RESEARCH ARTICLE Early Lineage Priming by Trisomy of Erg Leads to Myeloproliferation in a Down Syndrome Model Ashley P. Ng1,2*, Yifang Hu1, Donald Metcalf1,2, Craig D. Hyland1, Helen Ierino1, Belinda Phipson1,3,DiWu4,5, Tracey M. Baldwin1, Maria Kauppi1,2, Hiu Kiu1,2, Ladina Di Rago1, Douglas J. Hilton1,2, Gordon K. Smyth1,3, Warren S. Alexander1,2 1 The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria, Australia, 2 Department of Medical Biology, The University of Melbourne, Parkville, Victoria, Australia, 3 Department of Mathematics and Statistics, The University of Melbourne, Parkville, Victoria, Australia, 4 Centre for Cancer Research, Monash Institute of Medical Research, Monash University, Clayton, Victoria, Australia, 5 Department of Statistics, Harvard University, Cambridge, Massachusetts, United States of America * [email protected] OPEN ACCESS Abstract Citation: Ng AP, Hu Y, Metcalf D, Hyland CD, Ierino Down syndrome (DS), with trisomy of chromosome 21 (HSA21), is the commonest human H, Phipson B, et al. (2015) Early Lineage Priming by Trisomy of Erg Leads to Myeloproliferation in a Down aneuploidy. Pre-leukemic myeloproliferative changes in DS foetal livers precede the acqui- Syndrome Model. PLoS Genet 11(5): e1005211. sition of GATA1 mutations, transient myeloproliferative disorder (DS-TMD) and acute mega- doi:10.1371/journal.pgen.1005211 karyocytic leukemia (DS-AMKL). Trisomy of the Erg gene is required for myeloproliferation Editor: H. Leighton Grimes, Cincinnati Children's in the Ts(1716)65Dn DS mouse model. We demonstrate here that genetic changes specifi- Hospital Medical Center, UNITED STATES cally attributable to trisomy of Erg lead to lineage priming of primitive and early multipotential Received: September 3, 2014 progenitor cells in Ts(1716)65Dn mice, excess megakaryocyte-erythroid progenitors, and Accepted: April 13, 2015 malignant myeloproliferation. Gene expression changes dependent on trisomy of Erg in Ts (1716)65Dn multilineage progenitor cells were correlated with those associated with trisomy Published: May 14, 2015 of HSA21 in human DS hematopoietic stem and primitive progenitor cells. These data sug- Copyright: © 2015 Ng et al. This is an open access gest a role for ERG as a regulator of hematopoietic lineage potential, and that trisomy of article distributed under the terms of the Creative Commons Attribution License, which permits ERG in the context of DS foetal liver hemopoiesis drives the pre-leukemic changes that pre- unrestricted use, distribution, and reproduction in any dispose to subsequent DS-TMD and DS-AMKL. medium, provided the original author and source are credited. Data Availability Statement: All relevant data are within the paper and its Supporting Information files. All microarray files are available from the Array Author Summary Express database, www.ebi.ac.uk/arrayexpress An excess number of genes in trisomy on human chromosome 21 leads to the develop- (accession no. E-MTAB-2574). ment of specific diseases in human Down syndrome. An excess copy of the gene, ERG,an Funding: This work was supported by Program and ETS family transcription factor, has been implicated in abnormal blood system develop- Project Grants (1016647, 1054618), Fellowships ment in Down syndrome. In this study we show how trisomy of Erg in a murine Down (WSA 575501, GKS 1058892), and Independent syndrome model perturbs hematopoietic progenitor cells in a manner similar to that ob- Research Institutes Infrastructure Support Scheme Grant (361646) from the Australian National Health served in human Down syndrome by inducing gene expression changes and lineage prim- and Medical Research Council (https://www.nhmrc. ing in early multi-potential progenitors. We show that the gene expression signature gov.au), the Carden Fellowship (DM) of the Cancer specifically attributable to trisomy of Erg in the murine model is strongly correlated with Council, Victoria (http://www.cancervic.org.au), the PLOS Genetics | DOI:10.1371/journal.pgen.1005211 May 14, 2015 1/19 Genetic Consequences Erg Trisomy in Down Syndrome Cure Cancer Australia (www.cure.org.au/)/Leukaemia Foundation Australia (www.leukaemia.org.au/) Post gene expression changes in human Down syndrome hematopoietic cells. The data suggest Doctoral Fellowship and Lions Fellowship, Cancer that Erg is an important regulator of megakaryocyte-erythroid lineage specification in Council of Victoria (APN), the Australian Cancer multipotential hematopoietic cells and that trisomy of Erg in the context of DS prediposes Research Fund (http://acrf.com.au) and Victorian State Government Operational Infrastructure Support to a transient myeloproliferative disorder and acute megakaryocyte leukaemia in a multi- (www.vic.gov.au). The funders had no role in study step model of leukemogenesis. design, data collection and analysis, decision to publish, or preparation of the manuscript. Competing Interests: The authors have declared that no competing interests exist. Introduction Down syndrome (DS) is the commonest human aneuploidy [1]. DS infants with trisomy of human chromosome 21 (HSA21) are uniquely predisposed to a transient myeloproliferative disorder (DS-TMD) and acute megakaryocytic leukemia (DS-AMKL) [2]. DS-TMD, usually characterised by the presence of peripheral immature myeloblasts/megakaryoblasts and the variable involvement of other organs, is restricted to the neonatal period, spontaneously re- gresses and is the result of genetic co-operation between trisomy of HSA21 gene(s) with an ac- quired somatic mutation in GATA1 in virtually all cases [3]. However, up to 30% of children will subsequently develop DS-AMKL, a malignancy clonally related to the preceding DS-TMD. Candidate gene analysis and genome-wide exome sequencing have identified somatic muta- tions and deletions implicated in the progression of DS-TMD to DS-AMKL, in genes including JAK1, JAK2, JAK3, FLT3, TP53, TRIB1, MPL, EZH2, APC, PARK-2, PACRG, EXT1, DLEC1 and SMC3, and further suggested that GATA-1 mutations alone in the context of HSA21 triso- my were sufficient for development of DS-TMD [4–12]. Preceding acquisition of GATA1 mutations, human DS foetal livers exhibit perturbed hema- topoiesis. Increased numbers and clonogenicity of hematopoietic stem (HSC) and progenitor cells, increased frequency of bi-potential megakaryocyte-erythroid progenitors, and reduced numbers of granulocyte-macrophage-committed progenitor cells have been described [13–15]. This perturbation must be attributed to a specific trisomic gene or genes on HSA21 that drive the pre-leukemic DS phenotype from which DS-AMKL and DS-TMD subsequently arise. Mu- rine DS models with germline transmissible segmental trisomies of human or murine ortholo- gues of HSA21 genes have allowed genetic analyses of the contributions of genes within the DS critical interval to specific DS phenotypes [16–19]. A well studied model is the Ts(1716)65Dn mouse, which is trisomic for orthologs of ~104 human chromosome 21 genes [17]. Ts(1716) 65Dn mice display progressive myeloproliferation chracterised by thrombocytosis, megakaryo- cyte hyperplasia, dysplastic megakaryocytic morphology and myelofibrosis. Similarly, blasts with erythro-megakaryocytic features and myelofibrosis are commonly observed in organs af- fected by DS-TMD/AMKL, while DS foetal livers show increased numbers of bipotential mega- karyocyte-erythroid progenitors with increased clonogenicity and megakaryocyte/erythroid potential as well as megakaryocytosis [13–15]. We previously implicated the ETS family transcription factor ERG as a critical HSA21 gene in DS hematopoietic disease by demonstrating that specific reversion of Erg gene dosage to functional disomy, while the other ~103 orthologs remained trisomic, abrogated the myelo- megakaryocytic proliferation in Ts(1716)65Dn mice [20]. Erg has previously been shown to be essential for normal hematopoietic stem cell function [21–23]. Moreover, Erg deregulation can cause erythro-megakaryocytic leukemia in mice [24,25], and is implicated in acute myeloid and lymphoid malignancy in humans. In t(16;21) AML that carry the ERG/TLS-FUS fusion, complex karyotype AML with amplification of 21q, normal karyotype adult AML, MLL- rearranged paediatric AML and in T-ALL, high levels of ERG expression correlate with poor prognosis [26–29]. PLOS Genetics | DOI:10.1371/journal.pgen.1005211 May 14, 2015 2/19 Genetic Consequences Erg Trisomy in Down Syndrome The detailed mechanisms by which Erg contributes in trisomy to myeloproliferation in Ts (1716)65Dn mice, and whether molecular changes specifically driven by three copies of Erg in this model reflect
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