Abstract B6: Epigenetic regulation of Hedgehog pathway transcriptional output by BET bromodomain proteins

Aberrant activation of Hedgehog signaling drives oncogenesis in several types of cancer. As a result, there has been significant interest in developing therapeutic strategies targeting this pathway, most notably through inhibition of Smoothened. Though Smoothened inhibitors have shown efficacy in several cancer clinical trials, the initial enthusiasm for these inhibitors has been tempered by emergence of resistance and a priori resistance, often via mutation of Smoothened itself or through dysregulation of downstream components of the Hedgehog signaling axis. Here we reveal a strategy that overcomes these resistance mechanisms by targeting the far downstream transcriptional mediators of Hedgehog signaling through inhibition of the BET bromodomain protein, BRD4. We show that knockdown of BRD4 or treatment with the BET bromodomain inhibitor, JQ1, dramatically inhibits transcription of GLI1 and other Hedgehog target genes upon ligand-mediated or genetic activation of the Hedgehog pathway. We confirm the inhibitory effect of JQ1 occurs downstream of SMO and SUFU and verify by chromatin immunoprecipitation that Brd4 directly occupies the GLI1 and GLI2 promoters, with a substantial decrease in the engagement of these genomic sites upon treatment with JQ1. We observe a corresponding downregulation of genes associated with medulloblastoma-specific GLI1 binding sites upon exposure to JQ1, confirming the direct regulation of GLI1 by BET bromodomain proteins. Finally, using patient- and GEMM-derived cell lines of Hedgehog-driven cancer (basal cell carcinoma, medulloblastoma and ATRT), we show that JQ1 decreases Hh pathway output and proliferation, even in cell lines resistant to Smoothened inhibitors. These results expand the role of BET bromodomain inhibitors to targeting Hedgehog-driven cancers and highlight a strategy that overcomes the limitation of Hedgehog pathway inhibitors currently in clinical use. Citation Format: Yujie Tang, Simone Schubert, Jun Qi, Brian Nguyen, Sabran Masoud, Nujsaunusi Vue, Brianna Balansay, Furong Yu, Scott X. Atwood, Ramon J. Whitson, Anitha Ponnuswami, Spencer Chen, Sharareh Gholamin, Woo J. Pamelyn, Michelle Monje-Diesseroth, Sekyung Oh, Alex Lee, Jean Y. Tang, Rob Wechsler-Reya, Anthony E. Oro, James E. Bradner, Yoon-Jae Cho. Epigenetic regulation of Hedgehog pathway transcriptional output by BET bromodomain proteins. [abstract]. In: Proceedings of the AACR Special Conference on Pediatric Cancer at the Crossroads: Translating Discovery into Improved Outcomes; Nov 3-6, 2013; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2013;74(20 Suppl):Abstract nr B6.