title: MYCN transgenic zebrafish model with the characterization of acute myeloid leukemia and altered hematopoiesis.
abstract: BACKGROUND
  Amplification of MYCN (N-Myc) oncogene has been reported as a frequent event and a poor prognostic marker in human acute myeloid leukemia (AML). The molecular mechanisms and transcriptional networks by which MYCN exerts its influence in AML are largely unknown.
  
  
  RESULTS
  We introduced murine MYCN gene into embryonic zebrafish through a heat-shock promoter and established the stable germline Tg(MYCN:HSE:EGFP) zebrafish. N-Myc downstream regulated gene 1 (NDRG1), negatively controlled by MYCN in human and functionally involved in neutrophil maturation, was significantly under-expressed in this model. Using peripheral blood smear detection, histological section and flow cytometric analysis of single cell suspension from kidney and spleen, we found that MYCN overexpression promoted cell proliferation, enhanced the repopulating activity of myeloid cells and the accumulation of immature hematopoietic blast cells. MYCN enhanced primitive hematopoiesis by upregulating scl and lmo2 expression and promoted myelopoiesis by inhibiting gata1 expression and inducing pu.1, mpo expression. Microarray analysis identified that cell cycle, glycolysis/gluconeogenesis, MAPK/Ras, and p53-mediated apoptosis pathways were upregulated. In addition, mismatch repair, transforming and growth factor β (TGFβ) were downregulated in MYCN-overexpressing blood cells (p<0.01). All of these signaling pathways are critical in the proliferation and malignant transformation of blood cells.
  
  
  CONCLUSIONS
  The above results induced by overexpression of MYCN closely resemble the main aspects of human AML, suggesting that MYCN plays a role in the etiology of AML. MYCN reprograms hematopoietic cell fate by regulating NDRG1 and several lineage-specific hematopoietic transcription factors. Therefore, this MYCN transgenic zebrafish model facilitates dissection of MYCN-mediated signaling in vivo, and enables high-throughput scale screens to identify the potential therapeutic targets.
authors.affiliation: Department of Hematology, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
authors.forename: Li-Jing
authors.initials: LJ
authors.lastname: Shen
authors.forename: Fang-Yuan
authors.initials: FY
authors.lastname: Chen
authors.forename: Yong
authors.initials: Y
authors.lastname: Zhang
authors.forename: Lan-Fang
authors.initials: LF
authors.lastname: Cao
authors.forename: Ying
authors.initials: Y
authors.lastname: Kuang
authors.forename: Min
authors.initials: M
authors.lastname: Zhong
authors.forename: Ting
authors.initials: T
authors.lastname: Wang
authors.forename: Hua
authors.initials: H
authors.lastname: Zhong
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001773:Blood Cells / Q000473:pathology; D002453:Cell Cycle / Q000235:genetics; D019070:Cell Lineage / Q000235:genetics; D004195:Disease Models, Animal; D005260:Female; D018507:Gene Expression Regulation, Developmental; D053263:Gene Regulatory Networks; D016259:Genes, myc; D005822:Genetic Vectors; D006410:Hematopoiesis / Q000235:genetics*; D006412:Hematopoietic Stem Cells / Q000473:pathology; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000235:genetics* / Q000473:pathology; D008297:Male; D051379:Mice; D022423:Myeloid Cells / Q000378:metabolism / Q000473:pathology; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic; D015027:Zebrafish
chemical_list: D014157:Transcription Factors
references.citation: Nature. 2010 Mar 4;464(7285):112-5
references.pmid: 20154732
references.citation: Oncol Lett. 2012 May;3(5):1119-1123
references.pmid: 22783403
references.citation: Nat Protoc. 2009;4(1):44-57
references.pmid: 19131956
references.citation: Hematol Oncol. 2010 Jun;28(2):62-7
references.pmid: 20014148
references.citation: Development. 2000 May;127(9):1953-60
references.pmid: 10751183
references.citation: Cell. 2006 Jun 2;125(5):929-41
references.pmid: 16751102
references.citation: Proc Natl Acad Sci U S A. 2005 Apr 26;102(17):6068-73
references.pmid: 15827121
references.citation: J Biol Chem. 1993 Apr 15;268(11):8317-24
references.pmid: 8463341
references.citation: J Clin Invest. 2001 Sep;108(6):851-9
references.pmid: 11560954
references.citation: EMBO J. 2011 Aug 26;30(19):4059-70
references.pmid: 21873977
references.citation: Blood. 2010 Jan 28;115(4):824-33
references.pmid: 19965690
references.citation: Thromb Haemost. 1997 Feb;77(2):227-33
references.pmid: 9157571
references.citation: J Biol Chem. 2008 Jun 20;283(25):17652-61
references.pmid: 18413315
references.citation: Mech Dev. 2002 Oct;118(1-2):91-8
references.pmid: 12351173
references.citation: Cancer Res. 2007 Nov 15;67(22):10677-85
references.pmid: 18006809
references.citation: Blood. 2008 May 1;111(9):4690-9
references.pmid: 18305219
references.citation: Mol Cell Biol. 2008 Dec;28(24):7286-95
references.pmid: 18838534
references.citation: Int J Dev Biol. 2010;54(6-7):1127-37
references.pmid: 20711990
references.citation: Genes Cancer. 2010 Jun;1(6):605-16
references.pmid: 21779460
references.citation: Development. 2008 Jan;135(2):401-10
references.pmid: 18156164
references.citation: Genes Dev. 1998 Mar 1;12(5):621-6
references.pmid: 9499398
references.citation: Oncogene. 1998 Oct 22;17(16):2073-85
references.pmid: 9798678
references.citation: Br J Haematol. 1999 Dec;107(4):731-8
references.pmid: 10606877
references.citation: PLoS One. 2009 Nov 05;4(11):e7768
references.pmid: 19890398
references.citation: J Hepatol. 2008 Jun;48(6):962-73
references.pmid: 18384904
references.citation: Curr Opin Hematol. 2004 Jul;11(4):255-61
references.pmid: 15314525
references.citation: Blood. 2011 Feb 17;117(7):2146-56
references.pmid: 21076045
references.citation: Br J Haematol. 2011 Sep;154(5):590-9
references.pmid: 21749360
references.citation: J Biomed Sci. 2010 Mar 27;17(1):22
references.pmid: 20346173
references.citation: Leukemia. 2011 Feb;25(2):321-30
references.pmid: 21072044
references.citation: Dev Cell. 2005 Jan;8(1):97-108
references.pmid: 15621533
references.citation: Cell Stem Cell. 2008 Dec 4;3(6):611-24
references.pmid: 19041778
references.citation: Methods. 2001 Dec;25(4):402-8
references.pmid: 11846609
references.citation: Cell Cycle. 2007 May 15;6(10):1249-56
references.pmid: 17495526
references.citation: Leuk Res. 2010 Mar;34(3):393-8
references.pmid: 19775754
references.citation: Dev Biol. 2004 Jul 15;271(2):416-30
references.pmid: 15223344
references.citation: J Cell Biol. 1992 Jun;117(6):1137-50
references.pmid: 1607378
references.citation: Leuk Res. 2011 Jun;35(6):814-20
references.pmid: 21316758
references.citation: J Vis Exp. 2010 Mar 04;(37):
references.pmid: 20203557
references.citation: Cancer Res. 2006 Apr 15;66(8):4020-9
references.pmid: 16618721
references.citation: Blood. 2007 Mar 15;109(6):2389-98
references.pmid: 17090656
references.citation: Dev Biol. 2005 Jan 15;277(2):522-36
references.pmid: 15617691
references.citation: Genes Dev. 2004 Nov 15;18(22):2747-63
references.pmid: 15545632
references.citation: Leukemia. 2007 Mar;21(3):462-71
references.pmid: 17252014
references.citation: Leukemia. 2011 Nov;25(11):1739-50
references.pmid: 21701495
references.citation: Cell Cycle. 2006 Nov;5(22):2639-47
references.pmid: 17172827