Abstract A047: ONECUT2 is a targetable master regulator of aggressive variants of castration-resistant prostate cancer
Authors: Rotinen Mirja , You Sungyong , Yang Julie , Coetzee Simon , Huang Wen-Chin , Huang Fangjin , Pan Xinlei , Yáñez Alberto , Hazelett Dennis , Chu Chia-Yi , Chung Leland , Freedland Stephen , Vizio Dolores Di , Garraway Isla , Murali Ramachandran , Knudsen Beatrice , Freeman Michael 
Published: February 2019

Abstract
Background: Treatment of prostate cancer by hormone suppression leads to the appearance of aggressive variants of metastatic castration-resistant prostate cancer (mCRPC) with variable or no dependence on the androgen receptor (AR). Here we identify the neural transcription factor ONECUT2 as a negative regulator of the AR axis, that emerges in aggressive PC variants to control transcriptional networks linked to CRPC and neuroendocrine (NE) differentiation. We further demonstrate that ONECUT2 can be targeted with a small molecule that inhibits mCRPC metastasis in mice.
Methods: ONECUT2 was confirmed as a mCRPC-relevant protein and to be targetable by computational modeling and bioinformatics, enforced expression, silencing, microarray, ChIP-Seq, immunohistochemistry, immunofluorescence, quantitative imaging, functional assays, in vivo experiments, and surface plasmon resonance.
Results: We have performed a master regulator analysis using 260 mCRPC transcriptome profiles and developed a model transcription factor network for mCRPC that associates ONECUT2 for the first time with metastatic progression. Gene expression profiling of ONECUT2-engineered PC cell lines has allowed us to generate a ONECUT2 activity signature that reveals high positive correlation with pro-neural and aggressive PC signatures, and a negative correlation with AR activation pathways. We find that ONECUT2 is a negative regulator of AR expression and a repressor of its transcriptional program through direct binding to AR target genes. We also find that ONECUT2 is significantly increased in human NEPC and confers NE properties to CRPC through direct downregulation of the NEPC inhibitor FOXA1 and direct upregulation of the NEPC driver PEG10. Finally, we show that ONECUT2 is required for cell growth and survival and that it can be targeted with a small molecule that, by binding to the ONECUT2 C-terminal DNA binding domain, inhibits mCRPC metastasis in mice.
Conclusions: OC2 is a master regulator of aggressive mCRPC variants that drives AR-dependent adenocarcinoma toward NEPC differentiation by blocking AR/FOXA1-activity and inducing PEG10. OC2 can be targeted with a drug-like small molecule that inhibits CRPC metastasis in mice. Patients with OC2 active tumors may benefit from OC2 inhibitor therapy.
Citation Format: Mirja Rotinen, Sungyong You, Julie Yang, Simon Coetzee, Wen-Chin Huang, Fangjin Huang, Xinlei Pan, Alberto Yáñez, Dennis Hazelett, Chia-Yi Chu, Leland Chung, Stephen Freedland, Dolores Di Vizio, Isla Garraway, Ramachandran Murali, Beatrice Knudsen, Michael Freeman. ONECUT2 is a targetable master regulator of aggressive variants of castration-resistant prostate cancer [abstract]. In: Proceedings of the AACR Special Conference: Prostate Cancer: Advances in Basic, Translational, and Clinical Research; 2017 Dec 2-5; Orlando, Florida. Philadelphia (PA): AACR; Cancer Res 2018;78(16 Suppl):Abstract nr A047.