Inhibiting androgen receptor splice variants with cysteine-selective irreversible covalent inhibitors to treat prostate cancer.
Inhibiting androgen receptor splice variants with cysteine-selective irreversible covalent inhibitors to treat prostate cancer.
复制标题
DOI：
10.1073/pnas.2211832120
复制
发表时间：
2023-01-03
影响因子：
11.1
通讯作者：
中科院分区：
文献类型：
--
作者：
前列腺癌(PCA)在美国影响着超过25万名男性。雄激素受体(AR)信号转导抑制剂是治疗前列腺癌的主要药物。表达AR剪接变异体(AR-SVS)的高级PCA对当前的治疗策略没有反应。在这篇论文中，我们揭示了AR和AR-SV内在无序的反式激活结构域的物理化学性质，并开发了AR不可逆转的共价拮抗剂(SARICA)到反式激活结构域，用于治疗晚期PCa。SARICA还被用来进一步确定AR和AR-SV反式激活结构域中的潜在结合区域。雄激素受体(AR)及其剪接变异体(AR-SVS)通过转录重编程促进前列腺癌(PCa)的生长。AR和AR-SVS的低复杂性和内在无序的初级反式激活域(AF-1)调控PCa转录编程的机制尚不清楚。利用组学、细胞的活和固定荧光显微镜以及纯化的AF-1和AR-V7重组蛋白，我们在这里表明，AF-1和AR-V7剪接变体通过液-液相分离(LLP)形成分子凝聚体，表现出细胞内快速移动、共激活子相互作用和常染色质诱导等无序特征。LLP和其他无序特征被一类以UT-143为代表的小分子选择性AR不可逆共价拮抗剂(SARICA)逆转，它们与AF-1区域的C406和C327共价选择性结合。用UT-143干扰LLP的形成或突变导致染色质凝聚和AR-V7相互作用组的解离，最终形成转录不能的复合体。生化研究表明，AF-1区的C327和C406对于凝析油的形成、AR-V7的功能以及UT-143‘S不可逆的AR抑制是关键的。在治疗方面，UT-143具有类药物的药代动力学和代谢特性，并抑制前列腺癌细胞的增殖和肿瘤的生长。我们的工作提供了关键信息，表明临床上重要的AR-V7形成转录活性的分子缩合物，并与AF-1中的C327和C406共价结合，溶解缩合物，并抑制其功能。这项工作还确定了用于治疗前列腺癌的AF-1结合AR和AR-SV选择性共价抑制剂的文库。
Prostate cancer (PCa) affects over 250,000 men in the US. Androgen receptor (AR) signaling inhibitors are the mainstay therapeutics for PCa. Advanced PCa that expresses AR splice variants (AR-SVs) does not respond to current therapeutic strategies. In this manuscript, we uncovered the physicochemical properties of the intrinsically disordered transactivation domain of AR and AR-SV and developed AR-irreversible covalent antagonists (SARICA) to the transactivation domain for the treatment of advanced PCa. SARICAs were also used to identify a potential binding region in the AR and AR-SV transactivation domain. Androgen receptor (AR) and its splice variants (AR-SVs) promote prostate cancer (PCa) growth by orchestrating transcriptional reprogramming. Mechanisms by which the low complexity and intrinsically disordered primary transactivation domain (AF-1) of AR and AR-SVs regulate transcriptional programming in PCa remains poorly defined. Using omics, live and fixed fluorescent microscopy of cells, and purified AF-1 and AR-V7 recombinant proteins we show here that AF-1 and the AR-V7 splice variant form molecular condensates by liquid–liquid phase separation (LLPS) that exhibit disorder characteristics such as rapid intracellular mobility, coactivator interaction, and euchromatin induction. The LLPS and other disorder characteristics were reversed by a class of small-molecule-selective AR-irreversible covalent antagonists (SARICA) represented herein by UT-143 that covalently and selectively bind to C406 and C327 in the AF-1 region. Interfering with LLPS formation with UT-143 or mutagenesis resulted in chromatin condensation and dissociation of AR-V7 interactome, all culminating in a transcriptionally incompetent complex. Biochemical studies suggest that C327 and C406 in the AF-1 region are critical for condensate formation, AR-V7 function, and UT-143’s irreversible AR inhibition. Therapeutically, UT-143 possesses drug-like pharmacokinetics and metabolism properties and inhibits PCa cell proliferation and tumor growth. Our work provides critical information suggesting that clinically important AR-V7 forms transcriptionally competent molecular condensates and covalently engaging C327 and C406 in AF-1, dissolves the condensates, and inhibits its function. The work also identifies a library of AF-1-binding AR and AR-SV-selective covalent inhibitors for the treatment of PCa.
登录
查看更多内容
相位分离驱动异常的染色质循环和癌症发展。
影响因子：
64.8
作者：
Ahn JH;Davis ES;Daugird TA;Zhao S;Quiroga IY;Uryu H;Li J;Storey AJ;Tsai YH;Keeley DP;Mackintosh SG;Edmondson RD;Byrum SD;Cai L;Tackett AJ;Zheng D;Legant WR;Phanstiel DH;Wang GG
通讯作者：
Wang GG
影响因子：
7.7
作者：
Balbas MD;Evans MJ;Hosfield DJ;Wongvipat J;Arora VK;Watson PA;Chen Y;Greene GL;Shen Y;Sawyers CL
通讯作者：
Sawyers CL
影响因子：
4.8
作者：
Bohl, CE;Miller, DD;Dalton, JT
通讯作者：
Dalton, JT
影响因子：
8
作者：
Ahmed, Junaid;Meszaros, Attila;Tompa, Peter
通讯作者：
Tompa, Peter
影响因子：
50.3
作者：
Andersen, Raymond J.;Mawji, Nasrin R.;Sadar, Marianne D.
通讯作者：
Sadar, Marianne D.