title: WDR5 Supports an N-Myc Transcriptional Complex That Drives a Protumorigenic Gene Expression Signature in Neuroblastoma.
abstract: MYCN gene amplification in neuroblastoma drives a gene expression program that correlates strongly with aggressive disease. Mechanistically, trimethylation of histone H3 lysine 4 (H3K4) at target gene promoters is a strict prerequisite for this transcriptional program to be enacted. WDR5 is a histone H3K4 presenter that has been found to have an essential role in H3K4 trimethylation. For this reason, in this study, we investigated the relationship between WDR5-mediated H3K4 trimethylation and N-Myc transcriptional programs in neuroblastoma cells. N-Myc upregulated WDR5 expression in neuroblastoma cells. Gene expression analysis revealed that WDR5 target genes included those with MYC-binding elements at promoters such as MDM2. We showed that WDR5 could form a protein complex at the MDM2 promoter with N-Myc, but not p53, leading to histone H3K4 trimethylation and activation of MDM2 transcription. RNAi-mediated attenuation of WDR5 upregulated expression of wild-type but not mutant p53, an effect associated with growth inhibition and apoptosis. Similarly, a small-molecule antagonist of WDR5 reduced N-Myc/WDR5 complex formation, N-Myc target gene expression, and cell growth in neuroblastoma cells. In MYCN-transgenic mice, WDR5 was overexpressed in precancerous ganglion and neuroblastoma cells compared with normal ganglion cells. Clinically, elevated levels of WDR5 in neuroblastoma specimens were an independent predictor of poor overall survival. Overall, our results identify WDR5 as a key cofactor for N-Myc-regulated transcriptional activation and tumorigenesis and as a novel therapeutic target for MYCN-amplified neuroblastomas.
authors.affiliation: Children's Cancer Institute Australia, Randwick, Sydney, New South Wales, Australia.
authors.forename: Yuting
authors.initials: Y
authors.lastname: Sun
authors.affiliation: Institute of Molecular Medicine, Martin Luther University, ZAMED, Halle, Germany.
authors.forename: Jessica L
authors.initials: JL
authors.lastname: Bell
authors.affiliation: Children's Cancer Institute Australia, Randwick, Sydney, New South Wales, Australia.
authors.forename: Daniel
authors.initials: D
authors.lastname: Carter
authors.affiliation: Department of Pharmacy and Biotechnology, University of Bologna, Bologna, Italy.
authors.forename: Samuele
authors.initials: S
authors.lastname: Gherardi
authors.affiliation: Prince of Wales Clinical School and Lowy Cancer Research Centre, University of New South Wales Australia, Sydney, New South Wales, Australia.
authors.forename: Rebecca C
authors.initials: RC
authors.lastname: Poulos
authors.affiliation: Department of Pharmacy and Biotechnology, University of Bologna, Bologna, Italy.
authors.forename: Giorgio
authors.initials: G
authors.lastname: Milazzo
authors.affiliation: Prince of Wales Clinical School and Lowy Cancer Research Centre, University of New South Wales Australia, Sydney, New South Wales, Australia.
authors.forename: Jason W H
authors.initials: JW
authors.lastname: Wong
authors.affiliation: Drug Discovery Program, Ontario Institute for Cancer Research, Toronto, Ontario, Canada.
authors.forename: Rima
authors.initials: R
authors.lastname: Al-Awar
authors.affiliation: Children's Cancer Institute Australia, Randwick, Sydney, New South Wales, Australia.
authors.forename: Andrew E
authors.initials: AE
authors.lastname: Tee
authors.affiliation: Children's Cancer Institute Australia, Randwick, Sydney, New South Wales, Australia.
authors.forename: Pei Y
authors.initials: PY
authors.lastname: Liu
authors.affiliation: Children's Cancer Institute Australia, Randwick, Sydney, New South Wales, Australia.
authors.forename: Bing
authors.initials: B
authors.lastname: Liu
authors.affiliation: Children's Cancer Institute Australia, Randwick, Sydney, New South Wales, Australia.
authors.forename: Bernard
authors.initials: B
authors.lastname: Atmadibrata
authors.affiliation: Children's Cancer Institute Australia, Randwick, Sydney, New South Wales, Australia.
authors.forename: Matthew
authors.initials: M
authors.lastname: Wong
authors.affiliation: Children's Cancer Institute Australia, Randwick, Sydney, New South Wales, Australia.
authors.forename: Toby
authors.initials: T
authors.lastname: Trahair
authors.affiliation: School of Life Sciences, Nanjing University, Nanjing, Jiangsu, P.R. China.
authors.forename: Quan
authors.initials: Q
authors.lastname: Zhao
authors.affiliation: Department of Pediatrics, Section of Hematology-Oncology, Baylor College of Medicine, Houston, Texas.
authors.forename: Jason M
authors.initials: JM
authors.lastname: Shohet
authors.affiliation: Tumor Suppression Laboratory, Peter MacCallum Cancer Centre, East Melbourne, Victoria, Australia.
authors.forename: Ygal
authors.initials: Y
authors.lastname: Haupt
authors.affiliation: Department of Pediatric Oncology, Hematology and BMT, Charité University Medicine, Berlin, Germany. German Cancer Consortium (DKTK), Berlin, Germany. German Cancer Research Center (DKFZ), Heidelberg, Germany.
authors.forename: Johannes H
authors.initials: JH
authors.lastname: Schulte
authors.affiliation: Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Brown
authors.affiliation: Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada. Princess Margaret Cancer Centre, Toronto, Ontario, Canada.
authors.forename: Cheryl H
authors.initials: CH
authors.lastname: Arrowsmith
authors.affiliation: Structural Genomics Consortium, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Masoud
authors.initials: M
authors.lastname: Vedadi
authors.affiliation: Children's Cancer Institute Australia, Randwick, Sydney, New South Wales, Australia.
authors.forename: Karen L
authors.initials: KL
authors.lastname: MacKenzie
authors.affiliation: Institute of Molecular Medicine, Martin Luther University, ZAMED, Halle, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Hüttelmaier
authors.affiliation: Department of Pharmacy and Biotechnology, University of Bologna, Bologna, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Perini
authors.affiliation: Children's Cancer Institute Australia, Randwick, Sydney, New South Wales, Australia. Kids Cancer Centre, Sydney Children's Hospital, Randwick, New South Wales, Australia.
authors.forename: Glenn M
authors.initials: GM
authors.lastname: Marshall
authors.affiliation: Department of Pathology, University of Otago, Dunedin, New Zealand. Children's Medical Research Institute, University of Sydney, Sydney, New South Wales, Australia.
authors.forename: Antony
authors.initials: A
authors.lastname: Braithwaite
authors.affiliation: Children's Cancer Institute Australia, Randwick, Sydney, New South Wales, Australia. School of Women's and Children's Health, Faculty of Medicine, University of New South Wales, Randwick, Sydney, New South Wales, Australia. tliu@ccia.unsw.edu.au.
authors.forename: Tao
authors.initials: T
authors.lastname: Liu
mesh_terms: D000818:Animals; D063646:Carcinogenesis / Q000235:genetics; D048708:Cell Growth Processes / Q000235:genetics; D016259:Genes, myc; D057809:HEK293 Cells; D011495:Histone-Lysine N-Methyltransferase / Q000096:biosynthesis / Q000235:genetics*; D006657:Histones / Q000235:genetics / Q000378:metabolism; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D008745:Methylation; D051379:Mice; D008822:Mice, Transgenic; D009447:Neuroblastoma / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D051736:Proto-Oncogene Proteins c-mdm2 / Q000235:genetics; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics* / Q000378:metabolism; D051381:Rats; D014158:Transcription, Genetic; D059467:Transcriptome; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D015854:Up-Regulation
chemical_list: D006657:Histones; D047908:Intracellular Signaling Peptides and Proteins; C489427:MYC protein, human; D016271:Proto-Oncogene Proteins c-myc; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; C502547:WDR5 protein, human; D011495:Histone-Lysine N-Methyltransferase; C497206:MDM2 protein, human; D051736:Proto-Oncogene Proteins c-mdm2