title: CDK/CK1 inhibitors roscovitine and CR8 downregulate amplified MYCN in neuroblastoma cells.
abstract: To understand the mechanisms of action of (R)-roscovitine and (S)-CR8, two related pharmacological inhibitors of cyclin-dependent kinases (CDKs), we applied a variety of '-omics' techniques to the human neuroblastoma SH-SY5Y and IMR32 cell lines: (1) kinase interaction assays, (2) affinity competition on immobilized broad-spectrum kinase inhibitors, (3) affinity chromatography on immobilized (R)-roscovitine and (S)-CR8, (4) whole genome transcriptomics analysis and specific quantitative PCR studies, (5) global quantitative proteomics approach and western blot analysis of selected proteins. Altogether, the results show that the major direct targets of these two molecules belong to the CDKs (1,2,5,7,9,12), DYRKs, CLKs and CK1s families. By inhibiting CDK7, CDK9 and CDK12, these inhibitors transiently reduce RNA polymerase 2 activity, which results in downregulation of a large set of genes. Global transcriptomics and proteomics analysis converge to a central role of MYC transcription factors downregulation. Indeed, CDK inhibitors trigger rapid and massive downregulation of MYCN expression in MYCN-amplified neuroblastoma cells as well as in nude mice xenografted IMR32 cells. Inhibition of casein kinase 1 may also contribute to the antitumoral activity of (R)-roscovitine and (S)-CR8. This dual mechanism of action may be crucial in the use of these kinase inhibitors for the treatment of MYC-dependent cancers, in particular neuroblastoma where MYCN amplification is a strong predictor factor for high-risk disease.
authors: ManRos Therapeutics, Hôtel de Recherche, Centre de Perharidy, Roscoff, France.
authors: C
authors: C
authors: Delehouzé
authors: Evotec (München) GmbH, Am Klopferspitz 19a, Martinsried, Germany.
authors: K
authors: K
authors: Godl
authors: 1] ManRos Therapeutics, Hôtel de Recherche, Centre de Perharidy, Roscoff, France [2] C.N.R.S., 'Protein Phosphorylation & Human Disease' Group, Station Biologique, B.P. 74, Roscoff cedex, Bretagne, France.
authors: N
authors: N
authors: Loaëc
authors: ManRos Therapeutics, Hôtel de Recherche, Centre de Perharidy, Roscoff, France.
authors: C
authors: C
authors: Bruyère
authors: C.N.R.S., 'Protein Phosphorylation & Human Disease' Group, Station Biologique, B.P. 74, Roscoff cedex, Bretagne, France.
authors: N
authors: N
authors: Desban
authors: ManRos Therapeutics, Hôtel de Recherche, Centre de Perharidy, Roscoff, France.
authors: N
authors: N
authors: Oumata
authors: Laboratoire de Chimie Organique 2, INSERM U 648, Université Paris-Descartes, 4 avenue de l'Observatoire, Paris cedex, France.
authors: H
authors: H
authors: Galons
authors: Cancer Sciences & Clinical and Experimental Medicine, University of Southampton, Institute for Life Sciences, Center for Proteomics & Metabolomics Research, Highfield Campus, Southampton, UK.
authors: T I
authors: TI
authors: Roumeliotis
authors: Institute for Computational Biomedicine, Weil Cornell Medical College, New York, NY, USA.
authors: E G
authors: EG
authors: Giannopoulou
authors: Department of Tumor Cell Biology, St Jude Children's Research Hospital, 332 North Lauderdale, Memphis, TN, USA.
authors: J
authors: J
authors: Grenet
authors: Department of Tumor Cell Biology, St Jude Children's Research Hospital, 332 North Lauderdale, Memphis, TN, USA.
authors: D
authors: D
authors: Twitchell
authors: Department of Tumor Cell Biology, St Jude Children's Research Hospital, 332 North Lauderdale, Memphis, TN, USA.
authors: J
authors: J
authors: Lahti
authors: CNRS UMR 6290-Institut de Génétique et Développement de Rennes, Equipe Expression des Gènes et Oncogenèse, Université de Rennes1, SFR Biosit, Faculté de Médecine, 2 avenue du Pr. Léon Bernard, Rennes cedex, France.
authors: N
authors: N
authors: Mouchet
authors: CNRS UMR 6290-Institut de Génétique et Développement de Rennes, Equipe Expression des Gènes et Oncogenèse, Université de Rennes1, SFR Biosit, Faculté de Médecine, 2 avenue du Pr. Léon Bernard, Rennes cedex, France.
authors: M-D
authors: MD
authors: Galibert
authors: Cancer Sciences & Clinical and Experimental Medicine, University of Southampton, Institute for Life Sciences, Center for Proteomics & Metabolomics Research, Highfield Campus, Southampton, UK.
authors: S D
authors: SD
authors: Garbis
authors: 1] ManRos Therapeutics, Hôtel de Recherche, Centre de Perharidy, Roscoff, France [2] C.N.R.S., 'Protein Phosphorylation & Human Disease' Group, Station Biologique, B.P. 74, Roscoff cedex, Bretagne, France.
authors: L
authors: L
authors: Meijer
mesh_terms: D000818:Animals; D016203:CDC2 Protein Kinase / Q000037:antagonists & inhibitors; D045744:Cell Line, Tumor; D018844:Cyclin-Dependent Kinases / Q000037:antagonists & inhibitors; D015536:Down-Regulation / Q000187:drug effects; D005784:Gene Amplification; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D008822:Mice, Transgenic; D000071447:N-Myc Proto-Oncogene Protein; D009447:Neuroblastoma / Q000235:genetics* / Q000473:pathology; D009687:Nuclear Proteins / Q000235:genetics*; D015513:Oncogene Proteins / Q000235:genetics*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D011687:Purines / Q000494:pharmacology*; D011725:Pyridines / Q000494:pharmacology*; D000077546:Roscovitine; D023041:Xenograft Model Antitumor Assays
chemical_list: C532131:CR8 compound; C459801:MYCN protein, human; D000071447:N-Myc Proto-Oncogene Protein; D009687:Nuclear Proteins; D015513:Oncogene Proteins; D047428:Protein Kinase Inhibitors; D011687:Purines; D011725:Pyridines; D000077546:Roscovitine; D016203:CDC2 Protein Kinase; D018844:Cyclin-Dependent Kinases