Patients with castration-resistant prostate cancer inevitably acquire resistance to antiandrogen therapies in part because of androgen receptor (AR) mutations or splice variants enabling restored AR signaling. Here we show that ligand-activated AR can form transcriptionally active condensates. Both structured and unstructured regions of AR contribute to the effective phase separation of AR and disordered N-terminal domain plays a predominant role. AR liquid–liquid phase separation behaviors faithfully report transcriptional activity and antiandrogen efficacy. Antiandrogens can promote phase separation and transcriptional activity of AR-resistant mutants in a ligand-independent manner. We conducted a phase-separation-based phenotypic screen and identified ET516 that specifically disrupts AR condensates, effectively suppresses AR transcriptional activity and inhibits the proliferation and tumor growth of prostate cancer cells expressing AR-resistant mutants. Our results demonstrate liquid–liquid phase separation as an emerging mechanism underlying drug resistance and show that targeting phase separation may provide a feasible approach for drug discovery.

去势抵抗性前列腺癌患者不可避免地对抗雄激素疗法产生耐药性，部分原因是雄激素受体 (AR) 突变或剪接变异能够恢复 AR 信号。在这里，我们表明配体激活的 AR 可以形成转录活性凝聚物。AR 的结构化和非结构化区域都有助于 AR 的有效相分离，无序的 N 末端结构域起着主导作用。AR 液-液相分离行为忠实地报告了转录活性和抗雄激素功效。抗雄激素可以以不依赖配体的方式促进 AR 抗性突变体的相分离和转录活性。我们进行了基于相分离的表型筛选，并确定了 ET516 专门破坏 AR 凝聚物，有效抑制AR转录活性，抑制表达AR耐药突变体的前列腺癌细胞的增殖和肿瘤生长。我们的结果表明液-液相分离是一种新兴的耐药机制，并表明靶向相分离可能为药物发现提供一种可行的方法。