title: The Histone Methyltransferase DOT1L Promotes Neuroblastoma by Regulating Gene Transcription.
abstract: Myc oncoproteins exert tumorigenic effects by regulating expression of target oncogenes. Histone H3 lysine 79 (H3K79) methylation at Myc-responsive elements of target gene promoters is a strict prerequisite for Myc-induced transcriptional activation, and DOT1L is the only known histone methyltransferase that catalyzes H3K79 methylation. Here, we show that N-Myc upregulates DOT1L mRNA and protein expression by binding to the DOT1L gene promoter. shRNA-mediated depletion of DOT1L reduced mRNA and protein expression of N-Myc target genesODC1andE2F2DOT1L bound to the Myc Box II domain of N-Myc protein, and knockdown of DOT1L reduced histone H3K79 methylation and N-Myc protein binding at the ODC1 and E2F2 gene promoters and reduced neuroblastoma cell proliferation. Treatment with the small-molecule DOT1L inhibitor SGC0946 reduced H3K79 methylation and proliferation ofMYCNgene-amplified neuroblastoma cells. In mice xenografts of neuroblastoma cells stably expressing doxycycline-inducible DOT1L shRNA, ablating DOT1L expression with doxycycline significantly reduced ODC1 and E2F2 expression, reduced tumor progression, and improved overall survival. In addition, high levels of DOT1L gene expression in human neuroblastoma tissues correlated with high levels ofMYCN, ODC1, andE2F2gene expression and independently correlated with poor patient survival. Taken together, our results identify DOT1L as a novel cofactor in N-Myc-mediated transcriptional activation of target genes and neuroblastoma oncogenesis. Furthermore, they characterize DOT1L inhibitors as novel anticancer agents against MYCN-amplified neuroblastoma.Cancer Res; 77(9); 2522-33. ©2017 AACR.
authors.forename: Matthew
authors.initials: M
authors.lastname: Wong
authors.affiliation: Children's Cancer Institute Australia for Medical Research, University of New South Wales, Sydney, Australia.
authors.forename: Andrew E L
authors.initials: AEL
authors.lastname: Tee
authors.affiliation: Department of Pharmacy and Biotechnology, University of Bologna, Bologna, Italy.
authors.forename: Giorgio
authors.initials: G
authors.lastname: Milazzo
authors.affiliation: Institute of Molecular Medicine, Martin Luther University, ZAMED, Halle, Germany.
authors.forename: Jessica L
authors.initials: JL
authors.lastname: Bell
authors.affiliation: Prince of Wales Clinical School and Lowy Cancer Research Centre, University of New South Wales, Sydney, New South Wales, Australia.
authors.forename: Rebecca C
authors.initials: RC
authors.lastname: Poulos
authors.affiliation: Children's Cancer Institute Australia for Medical Research, University of New South Wales, Sydney, Australia.
authors.forename: Bernard
authors.initials: B
authors.lastname: Atmadibrata
authors.affiliation: Children's Cancer Institute Australia for Medical Research, University of New South Wales, Sydney, Australia.
authors.forename: Yuting
authors.initials: Y
authors.lastname: Sun
authors.affiliation: Children's Cancer Institute Australia for Medical Research, University of New South Wales, Sydney, Australia.
authors.forename: Duohui
authors.initials: D
authors.lastname: Jing
authors.affiliation: Children's Cancer Institute Australia for Medical Research, University of New South Wales, Sydney, Australia.
authors.forename: Nicholas
authors.initials: N
authors.lastname: Ho
authors.affiliation: Children's Cancer Institute Australia for Medical Research, University of New South Wales, Sydney, Australia.
authors.forename: Dora
authors.initials: D
authors.lastname: Ling
authors.affiliation: Children's Cancer Institute Australia for Medical Research, University of New South Wales, Sydney, Australia.
authors.forename: Pei Yan
authors.initials: PY
authors.lastname: Liu
authors.affiliation: School of Biomedical Sciences and Pharmacy, University of Newcastle, Newcastle, Australia.
authors.forename: Xu Dong
authors.initials: XD
authors.lastname: Zhang
authors.affiliation: Institute of Molecular Medicine, Martin Luther University, ZAMED, Halle, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Hüttelmaier
authors.affiliation: Prince of Wales Clinical School and Lowy Cancer Research Centre, University of New South Wales, Sydney, New South Wales, Australia.
authors.forename: Jason W H
authors.initials: JWH
authors.lastname: Wong
authors.affiliation: Children's Cancer Institute Australia for Medical Research, University of New South Wales, Sydney, Australia.|Centre for Childhood Cancer Research, UNSW Medicine, University of New South Wales, Sydney, New South Wales, Australia.
authors.forename: Jenny
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Pathology and Mechanisms of Disease and Translational Research, University of New South Wales, Sydney, New South Wales, Australia.
authors.forename: Patsie
authors.initials: P
authors.lastname: Polly
authors.affiliation: Department of Pharmacy and Biotechnology, University of Bologna, Bologna, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Perini
authors.affiliation: School of Environmental & Life Sciences, University of Newcastle, Ourimbah, New South Wales, Australia.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Scarlett
authors.affiliation: Children's Cancer Institute Australia for Medical Research, University of New South Wales, Sydney, Australia. tliu@ccia.unsw.edu.au.|Centre for Childhood Cancer Research, UNSW Medicine, University of New South Wales, Sydney, New South Wales, Australia.
authors.forename: Tao
authors.initials: T
authors.lastname: Liu
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D019175:DNA Methylation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000008:administration & dosage; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D000076983:Histone Methyltransferases; D011495:Histone-Lysine N-Methyltransferase / Q000037:antagonists & inhibitors / Q000235:genetics; D006801:Humans; D008780:Methyltransferases / Q000037:antagonists & inhibitors / Q000235:genetics*; D051379:Mice; D000071447:N-Myc Proto-Oncogene Protein / Q000096:biosynthesis / Q000235:genetics*; D009447:Neuroblastoma / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic
chemical_list: D004791:Enzyme Inhibitors; D000071447:N-Myc Proto-Oncogene Protein; C473477:DOT1L protein, human; D000076983:Histone Methyltransferases; D008780:Methyltransferases; D011495:Histone-Lysine N-Methyltransferase