title: MYCN and HDAC5 transcriptionally repress CD9 to trigger invasion and metastasis in neuroblastoma.
abstract: The systemic and resistant nature of metastatic neuroblastoma renders it largely incurable with current multimodal treatment. Clinical progression stems mainly from the increasing burden of metastatic colonization. Therapeutically inhibiting the migration-invasion-metastasis cascade would be of great benefit, but the mechanisms driving this cycle are as yet poorly understood. In-depth transcriptome analyses and ChIP-qPCR identified the cell surface glycoprotein, CD9, as a major downstream player and direct target of the recently described GRHL1 tumor suppressor. CD9 is known to block or facilitate cancer cell motility and metastasis dependent upon entity. High-level CD9 expression in primary neuroblastomas correlated with patient survival and established markers for favorable disease. Low-level CD9 expression was an independent risk factor for adverse outcome. MYCN and HDAC5 colocalized to the CD9 promoter and repressed transcription. CD9 expression diminished with progressive tumor development in the TH-MYCN transgenic mouse model for neuroblastoma, and CD9 expression in neuroblastic tumors was far below that in ganglia from wildtype mice. Primary neuroblastomas lacking MYCN amplifications displayed differential CD9 promoter methylation in methyl-CpG-binding domain sequencing analyses, and high-level methylation was associated with advanced stage disease, supporting epigenetic regulation. Inducing CD9 expression in a SH-EP cell model inhibited migration and invasion in Boyden chamber assays. Enforced CD9 expression in neuroblastoma cells transplanted onto chicken chorioallantoic membranes strongly reduced metastasis to embryonic bone marrow. Combined treatment of neuroblastoma cells with HDAC/DNA methyltransferase inhibitors synergistically induced CD9 expression despite hypoxic, metabolic or cytotoxic stress. Our results show CD9 is a critical and indirectly druggable suppressor of the invasion-metastasis cycle in neuroblastoma.
authors: Clinical Cooperation Unit Pediatric Oncology, German Cancer Research Center (DKFZ) and German Consortium for Translational Cancer Research (DKTK), INF, Heidelberg, Germany.|Phenex Pharmaceuticals AG, Waldhofer Straße, Heidelberg, Germany.
authors: Johannes
authors: J
authors: Fabian
authors: Clinical Cooperation Unit Pediatric Oncology, German Cancer Research Center (DKFZ) and German Consortium for Translational Cancer Research (DKTK), INF, Heidelberg, Germany.
authors: Desirée
authors: D
authors: Opitz
authors: Department of Pediatric Hematology and Oncology, University Children's Hospital Essen, Essen, Germany.
authors: Kristina
authors: K
authors: Althoff
authors: Clinical Cooperation Unit Pediatric Oncology, German Cancer Research Center (DKFZ) and German Consortium for Translational Cancer Research (DKTK), INF, Heidelberg, Germany.|Department of Pediatric Hematology, Oncology and SCT, Charité - University Hospital Berlin, Campus Virchow-Klinikum, Berlin, Germany.
authors: Marco
authors: M
authors: Lodrini
authors: Department of Pediatric Hematology and Oncology, University of Cologne, Cologne, Germany.
authors: Barbara
authors: B
authors: Hero
authors: Department of Pediatric Hematology and Oncology, University of Cologne, Cologne, Germany.
authors: Ruth
authors: R
authors: Volland
authors: Center for Medical Genetics Ghent, Ghent University, De Pintelaan, Ghent, Belgium.|Cancer Research Institute Ghent, De Pintelaan, Ghent, Belgium.
authors: Anneleen
authors: A
authors: Beckers
authors: Center for Medical Genetics Ghent, Ghent University, De Pintelaan, Ghent, Belgium.|Cancer Research Institute Ghent, De Pintelaan, Ghent, Belgium.
authors: Katleen
authors: K
authors: de Preter
authors: Center for Medical Genetics Ghent, Ghent University, De Pintelaan, Ghent, Belgium.|Cancer Research Institute Ghent, De Pintelaan, Ghent, Belgium.
authors: Anneleen
authors: A
authors: Decock
authors: Department of Experimental Surgery, Medical Faculty Mannheim, University of Heidelberg, Centre for Biomedicine and Medical Technology, Mannheim, Germany.
authors: Nitin
authors: N
authors: Patil
authors: Department of Experimental Surgery, Medical Faculty Mannheim, University of Heidelberg, Centre for Biomedicine and Medical Technology, Mannheim, Germany.
authors: Mohammed
authors: M
authors: Abba
authors: Department of Biostatistics, German Cancer Research Center (DKFZ), INF, Heidelberg, Germany.
authors: Annette
authors: A
authors: Kopp-Schneider
authors: Department of Pediatric Hematology, Oncology and SCT, Charité - University Hospital Berlin, Campus Virchow-Klinikum, Berlin, Germany.
authors: Kathy
authors: K
authors: Astrahantseff
authors: Clinical Cooperation Unit Pediatric Oncology, German Cancer Research Center (DKFZ) and German Consortium for Translational Cancer Research (DKTK), INF, Heidelberg, Germany.|Department of Pediatric Hematology, Oncology and SCT, Charité - University Hospital Berlin, Campus Virchow-Klinikum, Berlin, Germany.
authors: Jasmin
authors: J
authors: Wünschel
authors: Department of Pediatric Hematology, Oncology and SCT, Charité - University Hospital Berlin, Campus Virchow-Klinikum, Berlin, Germany.
authors: Sebastian
authors: S
authors: Pfeil
authors: Department of Pediatric Hematology, Oncology and SCT, Charité - University Hospital Berlin, Campus Virchow-Klinikum, Berlin, Germany.
authors: Maria
authors: M
authors: Ercu
authors: Department of Pediatric Hematology, Oncology and SCT, Charité - University Hospital Berlin, Campus Virchow-Klinikum, Berlin, Germany.
authors: Annette
authors: A
authors: Künkele
authors: Clinical Cooperation Unit Pediatric Oncology, German Cancer Research Center (DKFZ) and German Consortium for Translational Cancer Research (DKTK), INF, Heidelberg, Germany.|Yale University, New Haven, CT.
authors: Jamie
authors: J
authors: Hu
authors: Clinical Cooperation Unit Pediatric Oncology, German Cancer Research Center (DKFZ) and German Consortium for Translational Cancer Research (DKTK), INF, Heidelberg, Germany.|Department of Pediatric Hematology, Oncology and SCT, Charité - University Hospital Berlin, Campus Virchow-Klinikum, Berlin, Germany.
authors: Theresa
authors: T
authors: Thole
authors: Department of Neuropathology, University of Heidelberg, INF, Heidelberg, Germany.
authors: Leonille
authors: L
authors: Schweizer
authors: Department of Pathology, University of Heidelberg, INF, Heidelberg, Germany.
authors: Gunhild
authors: G
authors: Mechtersheimer
authors: Children's Cancer Institute, UNSW, Randwick, NSW, Australia.
authors: Daniel
authors: D
authors: Carter
authors: Children's Cancer Institute, UNSW, Randwick, NSW, Australia.
authors: Belamy B
authors: BB
authors: Cheung
authors: Division of Epigenomics and Cancer Risk Factors, DKFZ, INF, Heidelberg, Germany.
authors: Odilia
authors: O
authors: Popanda
authors: Department of Neuropathology, University of Heidelberg, INF, Heidelberg, Germany.|Clinical Cooperation Unit Neuropathology, DKFZ, INF, Heidelberg, Germany.
authors: Andreas
authors: A
authors: von Deimling
authors: Department of Oncogenomics and Emma Children's Hospital, Academic Medical Center, University of Amster- dam, Meibergdreef, Amsterdam, the Netherlands.
authors: Jan
authors: J
authors: Koster
authors: Department of Oncogenomics and Emma Children's Hospital, Academic Medical Center, University of Amster- dam, Meibergdreef, Amsterdam, the Netherlands.
authors: Rogier
authors: R
authors: Versteeg
authors: Neuroblastoma Genetics, DKFZ, INF, Heidelberg, Germany.
authors: Manfred
authors: M
authors: Schwab
authors: Children's Cancer Institute, UNSW, Randwick, NSW, Australia.|Kids Cancer Centre, Sydney Children's Hospital Randwick, Randwick, NSW, Australia.
authors: Glenn M
authors: GM
authors: Marshall
authors: Center for Medical Genetics Ghent, Ghent University, De Pintelaan, Ghent, Belgium.|Cancer Research Institute Ghent, De Pintelaan, Ghent, Belgium.
authors: Frank
authors: F
authors: Speleman
authors: Experimental Surgery and Tumor Cell Biology, University of Heidelberg, INF, Heidelberg, Germany.
authors: Ulrike
authors: U
authors: Erb
authors: Experimental Surgery and Tumor Cell Biology, University of Heidelberg, INF, Heidelberg, Germany.
authors: Margot
authors: M
authors: Zoeller
authors: Department of Experimental Surgery, Medical Faculty Mannheim, University of Heidelberg, Centre for Biomedicine and Medical Technology, Mannheim, Germany.
authors: Heike
authors: H
authors: Allgayer
authors: Department of Pediatric Hematology and Oncology, University of Cologne, Cologne, Ger-many.
authors: Thorsten
authors: T
authors: Simon
authors: Department of Pediatric Hematology and Oncology, University of Cologne, Cologne, Ger-many.|Center for Molecular Medicine Cologne, University of Cologne, Cologne, Germany.|Max Plank Institute for Metabolism Research, Cologne, Germany.
authors: Matthias
authors: M
authors: Fischer
authors: Department of Pediatric Hematology and Oncology, Heidelberg University, INF, Heidelberg, Germany.
authors: Andreas E
authors: AE
authors: Kulozik
authors: Department of Pediatric Hematology, Oncology and SCT, Charité - University Hospital Berlin, Campus Virchow-Klinikum, Berlin, Germany.
authors: Angelika
authors: A
authors: Eggert
authors: Clinical Cooperation Unit Pediatric Oncology, German Cancer Research Center (DKFZ) and German Consortium for Translational Cancer Research (DKTK), INF, Heidelberg, Germany.|Department of Pediatric Hematology and Oncology, Heidelberg University, INF, Heidelberg, Germany.
authors: Olaf
authors: O
authors: Witt
authors: Department of Pediatric Hematology, Oncology and SCT, Charité - University Hospital Berlin, Campus Virchow-Klinikum, Berlin, Germany.
authors: Johannes H
authors: JH
authors: Schulte
authors: Clinical Cooperation Unit Pediatric Oncology, German Cancer Research Center (DKFZ) and German Consortium for Translational Cancer Research (DKTK), INF, Heidelberg, Germany.|Department of Pediatric Hematology, Oncology and SCT, Charité - University Hospital Berlin, Campus Virchow-Klinikum, Berlin, Germany.|Department of Pediatric Hematology and Oncology, Heidelberg University, INF, Heidelberg, Germany.|Junior Neuroblastoma Research Group, Experimental and Clinical Research Center of the Max-Delbrück Center for Molecular Medicine in the Helmholtz Community and the Charité - University Medicine Berlin, Lindenberger Weg, Berlin, Germany.
authors: Hedwig E
authors: HE
authors: Deubzer
mesh_terms: D000818:Animals; D015972:Gene Expression Regulation, Neoplastic / Q000502:physiology*; D006655:Histone Deacetylases / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D000071447:N-Myc Proto-Oncogene Protein / Q000235:genetics / Q000378:metabolism*; D009361:Neoplasm Invasiveness / Q000235:genetics; D009447:Neuroblastoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D060245:Tetraspanin 29 / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D000071447:N-Myc Proto-Oncogene Protein; D060245:Tetraspanin 29; D006655:Histone Deacetylases
keywords: antimetastatic therapy; chromatin modulation; grainyhead-like transcription factor family; histone deacetylases; tetraspanin family