Gray Jaimie S, Wani Sajad A, Hussain Shahid, Huang Phoebe, Nayak Debasis, Long Mark D, Yates Clayton, Clinton Steven K, Bennet Chad E, Coss Christopher C, Campbell Moray J
College of Pharmacy, Division of Pharmaceutics and Pharmacology; The Ohio State University, Columbus, OH 43210.
College of Medicine; The Ohio State University, Columbus, OH 43210.
bioRxiv. 2023 Nov 17:2023.11.15.567282. doi: 10.1101/2023.11.15.567282.
We have dissected the role of Estrogen receptor beta (ERβ) in prostate cancer (PCa) with a novel ERβ ligand, OSU-ERb-12. Drug screens revealed additive interactions between OSU-ERB-12 and either epigenetic inhibitors or the androgen receptor antagonist, Enzalutamide (Enza). Clonogenic and cell biolody studies supported the potent additive effects of OSU-ERB-12 (100nM) and Enza (1μM). The cooperative behavior was in PCa cell lines treated with either OSU-ERB-12 plus Enza or combinations involving 17β-estradiol (E2). OSU-ERb-12 plus Enza uniquely impacted the transcriptiome, accessible chromatin, and the AR, MYC and H3K27ac cistromes. This included skewed transcriptional responses including suppression of the androgen and MYC transcriptomes, and repressed MYC protein. OSU-ERb-12 plus Enza uniquely impacted chromatin accessibility at approximately 3000 nucleosome-free sites, enriched at enhancers, enriched for basic Helix-Loop-Helix motifs. CUT&RUN experiments revealed combination treatment targeting of MYC, AR, and H3K27ac again shaping enhancer accessibility. Specifically, it repressed MYC interactions at enhancer regions enriched for bHLH motifs, and overlapped with publicly-available bHLH cistromes. Finally, cistrome-transcriptome analyses identified ~200 genes that distinguished advanced PCa tumors in the SU2C cohort with high androgen and low neuroendocrine scores.
我们使用新型雌激素受体β（ERβ）配体OSU-ERb-12剖析了ERβ在前列腺癌（PCa）中的作用。药物筛选显示OSU-ERB-12与表观遗传抑制剂或雄激素受体拮抗剂恩杂鲁胺（Enza）之间存在累加相互作用。克隆形成和细胞生物学研究支持了OSU-ERB-12（100nM）和Enza（1μM）的强效累加效应。在用OSU-ERB-12加Enza或涉及17β-雌二醇（E2）的组合处理的PCa细胞系中观察到了协同作用。OSU-ERb-12加Enza独特地影响转录组、可及染色质以及雄激素受体（AR）、MYC和H3K27ac染色质免疫沉淀测序峰（cistromes）。这包括转录反应的偏差，包括雄激素和MYC转录组的抑制以及MYC蛋白的抑制。OSU-ERb-12加Enza独特地影响了大约3000个无核小体位点的染色质可及性，这些位点在增强子处富集，富含碱性螺旋-环-螺旋基序。染色质切割与靶向测序（CUT&RUN）实验表明，联合处理对MYC、AR和H3K27ac的靶向作用再次塑造了增强子的可及性。具体而言，它抑制了在富含bHLH基序的增强子区域的MYC相互作用，并且与公开可用的bHLH染色质免疫沉淀测序峰重叠。最后，染色质免疫沉淀测序峰-转录组分析确定了约200个基因，这些基因可区分SU2C队列中具有高雄激素和低神经内分泌评分的晚期PCa肿瘤。
