Abstract 5815: The HBP1 tumor suppressor is a negative epigenetic regulator of MYCN driven neuroblastoma through interaction with the PRC2 complex
Authors: Denecker Geertrui , Claeys Shana , Lambertz Irina , Janssens Els , Vanhauwaert Suzanne , Decaesteker Bieke , Maerken Tom Van , Wilde Bram De , Laureys Genevieve , Althoff Kristina , Schulte Johannes , Demoulin Jean-Baptiste , Roberts Stephen S. , Bate-Eya Laurel , Molenaar Jan J. , Westermann Frank , Preter Katleen De , Speleman Frank 
Published: July 2017

Abstract
MYCN is a key driver in initiation and progression of neuroblastoma (NB) and represents a major target for novel drug strategies. We previously reported that the MYC repressor gene HBP1 was down regulated by mutant ALK via the PI3K-AKT-FOXO3 signaling axis1. Here, we further demonstrate that HBP1 upregulation suppresses proliferation of neuroblastoma cells by decreasing the MYCN signaling pathway. HBP1 levels were also shown to be repressed in neuroblastoma cells through MYC/MYCN driven upregulation of the miR-17~92 cluster, indicating that MYCN and mutant ALK both act to inhibit HBP1 expression in NB. Next, we tested the green tea polyphenol epigallocatechin gallate (EGCG), known to upregulate HBP1, and the BET inhibitor JQ1, which represses MYCN activity in neuroblastoma cells, and showed in vitro and in vivo synergistic effects on cell viability and tumor growth. Treatment with the PI3K/mTOR dual inhibitor BEZ-235 together with JQ1 also showed very strong synergistic effects. Further dissection of the HBP1 regulome using Gene Set Enrichment Analysis (GSEA) and iRegulon analysis (http://iregulon.aertslab.org) allowed identification of the PRC2 component SUZ12 as a central node in HBP1 regulated signaling, mainly through controlling the repression of MYCN regulated genes. In keeping with this finding, GSEA analysis of our HBP1 overexpression data set revealed also strong enrichment for genes that are differentially expressed upon EZH2 inhibition in neuroblastoma cells. Because HBP1 has previously been shown to interact with HDAC, we tested the effects of the HDAC inhibitors vorinostat and panobinostat as single agents and in combination with BEZ-235. Both combinations showed a strong synergistic effect on cell viability. The molecular mechanism of this synergism will be explored through RNAseq expression analysis. We conclude that HBP1 is a crucial component in MYCN controlled repression of gene activity through PRC2 interaction and demonstrate novel opportunities for precision drugging of MYCN overexpressing NB cells. 1Lambertz et al. Clin Cancer Res. (2015)
Citation Format: Geertrui Denecker, Shana Claeys, Irina Lambertz, Els Janssens, Suzanne Vanhauwaert, Bieke Decaesteker, Tom Van Maerken, Bram De Wilde, Genevieve Laureys, Kristina Althoff, Johannes Schulte, Jean-Baptiste Demoulin, Stephen S. Roberts, Laurel Bate-Eya, Jan J. Molenaar, Frank Westermann, Katleen De Preter, Frank Speleman. The HBP1 tumor suppressor is a negative epigenetic regulator of MYCN driven neuroblastoma through interaction with the PRC2 complex [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5815. doi:10.1158/1538-7445.AM2017-5815