Oral Presentation Advances in Neuroblastoma Research Congress 2016

MYCN and HDAC5 transcriptionally repress CD9 to trigger an invasion-metastasis cascade in neuroblastoma (#55)

Johannes Fabian 1 , Desiree Opitz 1 , Kristina Althoff 2 , Marco Lodrini 3 , Barbara Hero 4 , Ruth Volland 4 , Anneleen Beckers 5 , Katleen De Preter 5 , Nitin Patil 6 , Mohammed Abba 6 , Kathy Astrahantseff 3 , Jasmin Wünschel 3 , Maria Ercu 3 , Annette Kuenkele 3 , Jamie Hu 1 , Theresa Thole 3 , Schweizer Schweizer 7 , Gunhild Mechtersheimer 8 , Daniel Carter 9 , Belamy Cheung 9 , Odilia Popanda 10 , Andreas von Deimling 7 , Jan Koster 11 , Rogier Versteeg 11 , Kai-Oliver Henrich 12 , Frank Westermann 12 , Manfred Schwab 12 , Glenn Marshall 9 , Frank Spelemann 5 , Ulrike Erb 13 , Margot Zoeller 13 , Heike Allgayer 6 , Matthias Fischer 4 , Thorsten Simon 4 , Andreas Kulozik 14 , Angelika Eggert 3 , Olaf Witt 1 , Johannes Schulte 3 , Hedi Deubzer 3 15
  1. Clinical Cooperation Unit Pediatric Oncology, German Cancer Research Center (DKFZ), Heidelberg, Germany
  2. Department of Pediatric Hematology and Oncology, University Children’s Hospital Essen, Essen, Germany
  3. Charite, University Medicine Berlin, Berlin, BERLIN, Germany
  4. Department of Pediatric Hematology and Oncology, University of Cologne, Cologne, Germany
  5. Center for Medical Genetics Ghent (CMGG), Ghent University Hospital Medical Research Building (MRB), Ghent, Belgium
  6. Molecular Oncology of Solid Tumors Unit, German Cancer Research Center (DKFZ), Heidelberg, Germany
  7. Department of Neuropathology, Heidelberg University, Heidelberg, Germany
  8. Department of Pathology, Heidelberg University, Heidelberg, Germany
  9. Children’s Cancer Institute, UNSW, Randwick, Australia
  10. Division of Epigenomics and Cancer Risk Factors, German Cancer Research Center (DKFZ), Heidelberg, Germany
  11. Department of Oncogenomic, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands
  12. Neuroblastoma Genomics, German Cancer Research Center (DKFZ), Heidelberg, Germany
  13. Experimental Surgery and Tumor Cell Biology, Heidelberg University, Heidelberg, Germany
  14. Department of Pediatric Hematology and Oncology, Heidelberg University, Heidelberg, Germany
  15. Junior Neuroblastoma Research Group, Experimental and Clinical Research Center (ECRC), Berlin, Germany

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 displayed differential CD9 methylation in 450K methylation array analyses, and CD9 hypermethylation was associated with reduced CD9 expression, 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 mediator of neuroblastoma cell invasion and metastasis.