Abstract 5058: Silencing of NOTCH signaling enhances the sensitivity of ERG positive prostate cancer cells to AR inhibitors
Authors: Mohamed Ahmed A. , Tan Shyh-Han , Katta Shilpa , Xavier Charles P. , Ravindranath Lakshmi , Huang Wei , Li Hua , Srivastava Meera , Sharad Shashwat , Sreenath Taduru , Petrovics Gyorgy , Dobi Albert , Srivastava Shiv 
Published: August 2015

Introduction: Androgen receptor (AR) signaling plays a critical role in all the stages of prostate cancer (CaP) ranging from organ confined to castration-resistant (CRPC) phases. Although androgen deprivation therapy (ADT) remains the mainstay treatment for advanced CaP, the inevitable transition from androgen- sensitive to CRCP presents the most significant challenge in CaP therapy. Androgen dependent expression of oncogenic ETS related gene (ERG) in half of all CaP in western countries plays critical role in the tumorigenesis of CaP through regulation of cancer specific signaling pathways. We found that NOTCH transcription factors are common targets of ERG in ERG positive cancer cells. NOTCH signaling pathway is an important signaling pathway in the development of drug-resistant tumor growth. In the current study we evaluated the combinatorial effects of NOTCH and AR inhibitors in the context of ERG positive prostate cancer cells.

Methods: ERG, NOTCH1, NOTCH2 and downstream targets of NOTCH transcription factors were analyzed by Western blot assays. Dose and time kinetics of combining NOTCH inhibitor (γ-Secretase inhibitor 1, GSI-1) and AR inhibitors (Bicalutamide, Enzalutamide, and Abiraterone) were assessed in a panel of ERG positive or ERG negative CaP cells. Trypan blue exclusion, methylthiazole tetrazolium (MTT), or ApoTox-Glo™ Triplex assays were used to asses cell proliferation, apoptosis and drug cytotoxicity.

Results: Prostate cancer cell lines with endogenous or ectopic expression of ERG showed upregulation of NOTCH1 and NOTCH2. The NOTCH inhibitor, GSI-1 conferred an increased sensitivity to all tested AR inhibitors (Bicalutamide, Enzalutamide, and Abiraterone) with bicalutamide showing the most robust inhibition of AR, ERG, NOTCH1, NOTCH2, PSA, decreased cell growth and enhanced apoptosis in ERG positive VCaP cells. This observation was not seen in ERG negative LNCaP cells or in ERG positive primary endothelial cells.

Conclusions: NOTCH inhibitor enhanced sensitivity of AR inhibitors in ERG positive VCaP cells growth. The combination of the GSI-1 with AR inhibitors has shown synergistic effect when compared to single agent treatment. Taken together, our study suggests that NOTCH inhibitors may enhance the actions of AR inhibitors in the treatment of ERG positive prostate cancers. Inhibition of AR and NOTCH signaling may offer new opportunities in assessing ERG targeted therapy for prostate cancer.

Citation Format: Ahmed A. Mohamed, Shyh-Han Tan, Shilpa Katta, Charles P. Xavier, Lakshmi Ravindranath, Wei Huang, Hua Li, Meera Srivastava, Shashwat Sharad, Taduru Sreenath, Gyorgy Petrovics, Albert Dobi, Shiv Srivastava. Silencing of NOTCH signaling enhances the sensitivity of ERG positive prostate cancer cells to AR inhibitors. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 5058. doi:10.1158/1538-7445.AM2015-5058