尽管在控制使用 ADT/ARSI 难以治愈的前列腺癌 (PCa) 进展方面取得了一些进展，但大多数患者最终还是死于该疾病，并且迫切需要了解导致 CRPC 发展的机制。一个常见的机制是能够整合来自睾酮水平消失的 AR 信号，YAP 作为共激活剂频繁参与，并指出 Hippo 通路的放松管制是一个主要决定因素。我们最近表明，YAP 通过稳定 Y407 处的磷酸化，通过 TLK1>NEK1 轴进行转录后激活。我们现在通过展示以下内容来巩固这项工作：(1) Y407 的磷酸化对于 YAP 在细胞核中的保留/分配至关重要，而 J54 (TLK1i) 与 YAP-Y407 去磷酸化一起逆转了这一点。 (2) J54 增强了（细胞质）YAP 的降解，抵消了恩杂鲁胺诱导的积累。 (3) 所有这些效应（包括 YAP 核保留）的基础可以通过 pYAP-Y407 与其转录共激活因子 AR 和 TEAD1 更强的关联来解释。 (4) 我们证明，在典型 ARE 和 TEAD1 驱动基因的启动子处，对于 GFP-YAP-wt 的 ChIP 很容易检测到，但对于 GFP-YAP-Y407F 突变体却很难检测到，但在用 J54 处理后会被替换。 (5) 虽然 LNCaP 细胞的异种移植物在 ARSI J54 治疗后显示出快速消退，但在 VCaP 模型中，在 TMPRSS2-ERG 致癌易位的驱动下，肿瘤最初对组合反应良好，但随后复发，尽管 pNek1-T141 持续受到抑制和pYAP-Y407。这表明从长远来看，VCaP 肿瘤的 CRPC 进展有一条替代平行途径，该途径可能与观察到的 ENZ 驱动的 YAP 失调无关，尽管显然一些 YAP 基因靶标（如 PD-L1）在长期 ENZ 治疗后会积累，仍然受到同时添加 J54 的抑制。

Despite some advances in controlling the progression of prostate cancer (PCa) that is refractory to the use of ADT/ARSI, most patients eventually succumb to the disease, and there is a pressing need to understand the mechanisms that lead to the development of CRPC. A common mechanism is the ability to integrate AR signals from vanishing levels of testosterone, with the frequent participation of YAP as a co-activator, and pointing to the deregulation of the Hippo pathway as a major determinant. We have recently shown that YAP is post-transcriptionally activated via the TLK1>NEK1 axis by stabilizing phosphorylation at Y407. We are now solidifying this work by showing the following: (1) The phosphorylation of Y407 is critical for YAP retention/partition in the nuclei, and J54 (TLK1i) reverses this along with YAP-Y407 dephosphorylation. (2) The enhanced degradation of (cytoplasmic) YAP is increased by J54 counteracting its Enzalutamide-induced accumulation. (3) The basis for all these effects, including YAP nuclear retention, can be explained by the stronger association of pYAP-Y407 with its transcriptional co-activators, AR and TEAD1. (4) We demonstrate that ChIP for GFP-YAP-wt, but hardly for the GFP-YAP-Y407F mutant, at the promoters of typical ARE- and TEAD1-driven genes is readily detected but becomes displaced after treatment with J54. (5) While xenografts of LNCaP cells show rapid regression following treatment with ARSI+J54, in the VCaP model, driven by the TMPRSS2-ERG oncogenic translocation, tumors initially respond well to the combination but subsequently recur, despite the continuous suppression of pNek1-T141 and pYAP-Y407. This suggests an alternative parallel pathway for CRPC progression for VCaP tumors in the long term, which may be separate from the observed ENZ-driven YAP deregulation, although clearly some YAP gene targets like PD-L1, that are found to accumulate following prolonged ENZ treatment, are still suppressed by the concomitant addition of J54.