Jonas Ecker1,2,3, Venu Thatikonda1,4, Gianluca Sigismondo5, Florian Selt1,3,6, Gintvile Valinciute1,3,7, Ina Oehme1,3, Carina Müller1,3, Juliane L Buhl1,3,7, Johannes Ridinger1,3, Diren Usta1,3, Nan Qin8,9, Cornelis M van Tilburg1,2,3, Christel Herold-Mende10, Marc Remke8,9, Felix Sahm1,11, Frank Westermann1,3,12, Marcel Kool1,4, Robert J Wechsler-Reya13, Lukas Chavez14, Jeroen Krijgsveld5,15, Natalie Jäger4, Stefan M Pfister1,2,4, Olaf Witt1,2,3, Till Milde1,2,3. 1. Hopp Children's Cancer Center Heidelberg (KiTZ), Heidelberg, Germany. 2. KiTZ Clinical Trial Unit, Department of Pediatric Hematology and Oncology, Heidelberg University Hospital, Heidelberg, Germany. 3. Clinical Cooperation Unit Pediatric Oncology, German Cancer Research Center and German Consortium for Translational Cancer Research, Heidelberg, Germany. 4. Division of Pediatric Neurooncology, German Cancer Research Center and German Consortium for Translational Cancer Research, Heidelberg, Germany. 5. Division of Proteomics of Stem Cells and Cancer, German Cancer Research Center, Heidelberg, Germany. 6. Clinical Cooperation Unit Neuropathology, German Consortium for Translational Cancer Research, German Cancer Research Center, Heidelberg, Germany. 7. Faculty of Biosciences, Heidelberg University, Heidelberg, Germany. 8. Department of Pediatric Oncology, Hematology, and Clinical Immunology, Medical Faculty, University Hospital Düsseldorf, Germany. 9. Department of Pediatric Neuro-Oncogenomics, German Cancer Consortium and German Cancer Research Center, Heidelberg, Germany. 10. Department of Neurosurgery, Heidelberg University Hospital, Heidelberg, Germany. 11. Department of Neuropathology, Institute of Pathology, University Hospital, Heidelberg, Germany. 12. Division of Neuroblastoma Genomics, German Cancer Research Center, Heidelberg, Germany. 13. Tumor Initiation and Maintenance Program, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, California, USA. 14. Department of Medicine, University of California San Diego, San Diego, California, USA. 15. Heidelberg University, Medical Faculty, Heidelberg, Germany.
Abstract
BACKGROUND: The sensitivity of myelocytomatosis oncogene (MYC) amplified medulloblastoma to class I histone deacetylase (HDAC) inhibition has been shown previously; however, understanding the underlying molecular mechanism is crucial for selection of effective HDAC inhibitors for clinical use. The aim of this study was to investigate the direct molecular interaction of MYC and class I HDAC2, and the impact of class I HDAC inhibition on MYC function. METHODS: Co-immunoprecipitation and mass spectrometry were used to determine the co-localization of MYC and HDAC2. Chromatin immunoprecipitation (ChIP) sequencing and gene expression profiling were used to analyze the co-localization of MYC and HDAC2 on DNA and the impact on transcriptional activity in primary tumors and a MYC amplified cell line treated with the class I HDAC inhibitor entinostat. The effect on MYC was investigated by quantitative real-time PCR, western blot, and immunofluorescence. RESULTS: HDAC2 is a cofactor of MYC in MYC amplified medulloblastoma. The MYC-HDAC2 complex is bound to genes defining the MYC-dependent transcriptional profile. Class I HDAC inhibition leads to stabilization and reduced DNA binding of MYC protein, inducing a downregulation of MYC activated genes (MAGs) and upregulation of MYC repressed genes (MRGs). MAGs and MRGs are characterized by opposing biological functions and by distinct enhancer-box distribution. CONCLUSIONS: Our data elucidate the molecular interaction of MYC and HDAC2 and support a model in which inhibition of class I HDACs directly targets MYC's transactivating and transrepressing functions.
BACKGROUND: The sensitivity of myelocytomatosis oncogene (MYC) amplified medulloblastoma to class I histone deacetylase (HDAC) inhibition has been shown previously; however, understanding the underlying molecular mechanism is crucial for selection of effective HDAC inhibitors for clinical use. The aim of this study was to investigate the direct molecular interaction of MYC and class I HDAC2, and the impact of class I HDAC inhibition on MYC function. METHODS: Co-immunoprecipitation and mass spectrometry were used to determine the co-localization of MYC and HDAC2. Chromatin immunoprecipitation (ChIP) sequencing and gene expression profiling were used to analyze the co-localization of MYC and HDAC2 on DNA and the impact on transcriptional activity in primary tumors and a MYC amplified cell line treated with the class I HDAC inhibitor entinostat. The effect on MYC was investigated by quantitative real-time PCR, western blot, and immunofluorescence. RESULTS:HDAC2 is a cofactor of MYC in MYC amplified medulloblastoma. The MYC-HDAC2 complex is bound to genes defining the MYC-dependent transcriptional profile. Class I HDAC inhibition leads to stabilization and reduced DNA binding of MYC protein, inducing a downregulation of MYC activated genes (MAGs) and upregulation of MYC repressed genes (MRGs). MAGs and MRGs are characterized by opposing biological functions and by distinct enhancer-box distribution. CONCLUSIONS: Our data elucidate the molecular interaction of MYC and HDAC2 and support a model in which inhibition of class I HDACs directly targets MYC's transactivating and transrepressing functions.
Authors: Manpreet Kalkat; Diana Resetca; Corey Lourenco; Pak-Kei Chan; Yong Wei; Yu-Jia Shiah; Natasha Vitkin; Yufeng Tong; Maria Sunnerhagen; Susan J Done; Paul C Boutros; Brian Raught; Linda Z Penn Journal: Mol Cell Date: 2018-11-08 Impact factor: 17.970
Authors: Joanna Triscott; Cathy Lee; Colleen Foster; Branavan Manoranjan; Mary Rose Pambid; Rachel Berns; Abbas Fotovati; Chitra Venugopal; Katrina O'Halloran; Aru Narendran; Cynthia Hawkins; Vijay Ramaswamy; Eric Bouffet; Michael D Taylor; Ash Singhal; Juliette Hukin; Rod Rassekh; Stephen Yip; Paul Northcott; Sheila K Singh; Christopher Dunham; Sandra E Dunn Journal: Cancer Res Date: 2013-09-09 Impact factor: 12.701
Authors: Yanxin Pei; Kun-Wei Liu; Jun Wang; Alexandra Garancher; Ran Tao; Lourdes A Esparza; Donna L Maier; Yoko T Udaka; Najiba Murad; Sorana Morrissy; Huriye Seker-Cin; Sebastian Brabetz; Lin Qi; Mari Kogiso; Simone Schubert; James M Olson; Yoon-Jae Cho; Xiao-Nan Li; John R Crawford; Michael L Levy; Marcel Kool; Stefan M Pfister; Michael D Taylor; Robert J Wechsler-Reya Journal: Cancer Cell Date: 2016-03-14 Impact factor: 31.743
Authors: Jonas Ecker; Ina Oehme; Ralph Mazitschek; Andrey Korshunov; Marcel Kool; Thomas Hielscher; Judit Kiss; Florian Selt; Carina Konrad; Marco Lodrini; Hedwig E Deubzer; Andreas von Deimling; Andreas E Kulozik; Stefan M Pfister; Olaf Witt; Till Milde Journal: Acta Neuropathol Commun Date: 2015-04-03 Impact factor: 7.801
Authors: Florian Selt; Juliane Hohloch; Thomas Hielscher; Felix Sahm; David Capper; Andrey Korshunov; Diren Usta; Sebastian Brabetz; Johannes Ridinger; Jonas Ecker; Ina Oehme; Jan Gronych; Viktoria Marquardt; David Pauck; Heidi Bächli; Charles D Stiles; Andreas von Deimling; Marc Remke; Martin U Schuhmann; Stefan M Pfister; Tilman Brummer; David T W Jones; Olaf Witt; Till Milde Journal: Oncotarget Date: 2017-02-14
Authors: Margaret Shatara; Kathleen M Schieffer; Darren Klawinski; Diana L Thomas; Christopher R Pierson; Eric A Sribnick; Jeremy Jones; Diana P Rodriguez; Carol Deeg; Elizabeth Hamelberg; Stephanie LaHaye; Katherine E Miller; James Fitch; Benjamin Kelly; Kristen Leraas; Ruthann Pfau; Peter White; Vincent Magrini; Richard K Wilson; Elaine R Mardis; Mohamed S Abdelbaki; Jonathan L Finlay; Daniel R Boué; Catherine E Cottrell; David R Ghasemi; Kristian W Pajtler; Diana S Osorio Journal: Acta Neuropathol Commun Date: 2021-12-11 Impact factor: 7.801