Background: Inflammation, mediated by NF-κB, is linked to prostate cancer (PCa) progression and is commonly observed in castrate-resistant disease. We have previously shown that low expression of tristetraprolin (TTP), which modulates the NF-κB pathway and inflammation, is prognostic for lethal PCa after surgery. It has been suggested that NF-κB-driven resistance in PCa cells may be mediated by aberrant androgen receptor (AR) activation and AR splice variant production. Reversal of resistance to androgen deprivation therapy (ADT) may therefore be achieved by utilizing NF-κB inhibitors, such as dimethylaminoparthenolide (DMAPT). Materials and Methods: To examine clinical correlations between TTP/NF-κB and AR-V7, gene-expression profiling data and clinical data was obtained from several publicly available PCa cohorts (TCGA, SU2C, Taylor et. al.). Quantitative immunofluorescence for TTP and PTEN was performed on tissue microarrays from a clinical cohort of patients treated by radical prostatectomy with known time to biochemical recurrence (BCR). In vitro, mouse (MYC-CaP) and human (VCaP-CR, LNCaP-95) PCa cell lines were treated with TNF-α (20 ng/mL) to stimulate NF-κB expression, DMAPT (5-10 μM) to inhibit NF-κB and Enzalutamide (10 μM) in various combinations. Cell lines stably expressing the IκBα super repressor (MUT IκBα) were generated for comparison to DMAPT-mediated inhibition of NF-κB. Expression of AR and AR variants was measured by western blot and qPCR. For in vivo experiments, SCID mice bearing VCaP-CR xenograft tumors were treated with surgical castration, DMAPT (100 mg/kg, oral gavage, QD), surgical castration plus DMAPT or water vehicle control (n = 7-8 per group). Xenograft tumors were measured thrice weekly until they reached an ethical endpoint or after 80 days of treatment. Time-to-progression of tumors was assessed by Kaplan-Meier analysis and molecular differences within tumors analyzed by qPCR and immunohistochemistry (IHC). Results: In multiple clinical PCa cohorts, reduced TTP expression correlated with an increased AR-V7 signature. In a cohort of patients with localized PCa, decreased TTP expression reduced time to BCR and co-loss of TTP and PTEN (a known PCa tumor suppressor) further decreased time to BCR. Activation of NF-κB in PCa cell lines resulted in a significant increase in AR variant expression and minor upregulation of AR. Chemical (DMAPT) and genetic (MUT IκBα) inhibition of NF-κB slowed tumor proliferation and decreased AR variant expression. Treatment of PCa cell lines with Enzalutamide resulted in AR variant upregulation, which was blocked by DMAPT. Castration of mice bearing VCaP-CR tumors significantly increased AR and AR-V7 tumor expression. DMAPT treatment reduced NF-κB and AR-V7 expression in xenograft tumors, but did not significantly alter tumor growth compared to vehicle treatment. The combination of castration and DMAPT slowed median time-to-progression of xenograft tumors compared to vehicle-treated tumors (79 and 36 days respectively, p < 0.001). Combination therapy decreased Ki-67 (IHC), AR-V7 and AR (IHC and qPCR) expression in xenograft tumors. Conclusions: Current treatments for PCa mainly target AR; however, despite initial response, these treatments commonly fail. Our data highlight the relationship between NF-κB and AR variant expression in aggressive PCa and support the rationale for clinical use of DMAPT to treat castrate-resistant tumors in combination with AR antagonists.
Citation Format: Katherine L Morel, Anis A Hamid, Leigh Ellis, Christopher J Sweeney. Dimethylaminoparthenolide-mediated inhibition of NF-κB decreases resistance to ADT by targeting AR variants in lethal prostate cancer [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr B038. doi:10.1158/1535-7163.TARG-19-B038