Abstract B049: DNA damage echoes are key to prostate cancer susceptibility to supraphysiologic androgen levels
Authors: Chatterjee Payel , Lucas Jared M. , Nyquist Michael D. , Corey Eva , Nelson Peter S. 
Published: February 2019

Purpose: Prostate cancer cells are dependent on AR for growth and survival, but recent reports have shown a paradoxical role of supraphysiologic androgen in growth inhibition, known as hormone interference. Androgen stimulation in AR-overexpressing prostate cancer cells leads to formation of double-strand breaks (DSBs). Here, we investigate the potential mechanism of high androgen induced DSBs and growth inhibition and its effect on survival following androgen stimulation in combination with DNA damage repair inhibitors or defects.

Experimental Design: AR-dependent LNCaP, AR overexpressing LNCaP and PC3, AR-independent PC3, and APIPC were used for this study. High levels of androgen (R1881)/ DHT and olaparib-induced DNA damage were evaluated by gH2AX and 53BP1 foci formation after 6 or 24h. For cell death assays the drug was administered continuously for 3 days following the plating day. DNA PKcs activity was determined by Ser2056foci formation and dissociation from chromatin was measured by Thr2609 foci. AR activity or DNA binding was evaluated by Ser81 AR phosphorylation. An ex vivo PDX model (LuCaP 96CR) was utilized to mimic the BRCA2 defect condition.

Results: We demonstrated that supraphysiologic androgen treatment on androgen-dependent and AR-overexpressing prostate cancer cell lines resulted in a dose-dependent induction of widespread DSBs, which was augmented after PARP inhibition (olaparib). AR expression correlated with persistent DNA damage as AR-independent cells lacked DSBs. High androgen treatment significantly induced caspase activity and reduced survival in AR-overexpressing cells. Hyperactivated DNA PKcs was retained on chromatin following high androgen exposure in these cells, leading to impaired dissociation of DNA PKcs from chromatin and inducing DNA damage. AR phosphorylation at Ser81 was also in concordance with hyperphosphorylated DNA PKcs, particularly in AR-overexpressing cells following high androgen and olaparib treatment. HR-deficient, BRCA2-null cells induced more DSBs after high androgen treatment with or without olaparib and were susceptible to DNA PKcs inhibition following androgen treatment. In contrast to this, AR-overexpressing cells did not exhibit any DNA damage following DNA PKcs inhibition after high androgen exposure. Ex vivo PDX models demonstrated a synergistic interaction between olaparib and high levels of androgen-induced DSB formation as well as hyperactive DNA PKcs.

Conclusions: These results suggest a potential mechanism of supraphysiologic androgen stimulated AR-induced DNA damage and proliferation inhibition. AR binds to DNA PKcs following high androgen and until AR levels become saturated DNA PKcs and AR stay on the chromatin, which leads to inefficient DNA damage repair. HR-deficient cells were particularly susceptible to high androgen. Our data provide a mechanistic rationale for the response of AR upregulated cells as well as DNA repair deficient cells to supraphysiologic androgen therapy in combination with PARP inhibition.

Citation Format: Payel Chatterjee, Jared M. Lucas, Michael D. Nyquist, Eva Corey, Peter S. Nelson. DNA damage echoes are key to prostate cancer susceptibility to supraphysiologic androgen levels [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 B049.