细胞色素P450（CYP）基因的诱导是药物间相互作用的一个重要原因，对新药候选化合物进行诱导倾向的临床前评估是强制性的。YAP/TEAD信号传导已成为各种肿瘤治疗适应症的有吸引力的靶点，多种化学结构不同的YAP/TEAD抑制剂正在快速进入临床阶段。在这里，我们测试了一个多样的YAP/TEAD抑制剂集合的CYP诱导倾向，它们具有不同的作用方式和TEAD isoform 选择性在原代人肝细胞（PHH）的单层和3D球体中。我们发现，YAP/TEAD抑制导致2D单层中CYP广泛诱导，而在球体培养中仅有较少的诱导。综合RNA-Seq结果表明，与球体相比，YAP/TEAD信号在2D培养中增加，这与相互作用的转录因子LXR和ESRRA的活性升高相一致，可能至少部分是由于机械感受的改变。抑制此YAP/TEAD高活化导致肝细胞分化减少、肝功能增加，包括CYP。因此，这些结果表明观察到的诱导是化合物的靶向效应，而不是对异物感受核受体的直接激活。综上所述，所呈现的数据将肝细胞分化减少与YAP/TEAD失调联系起来，揭示了一种新的非经典CYP诱导途径，并凸显了器官模拟的3D培养用于预测临床相关药代动力学特性的优势，尤其是对于非典型的诱导机制。 版权所有© 2023 The Author(s). 由 Elsevier Inc.发表保留所有权利。

Induction of cytochrome P450 (CYP) genes constitutes an important cause of drug-drug interactions and preclinical evaluation of induction liability is mandatory for novel drug candidates. YAP/TEAD signaling has emerged as an attractive target for various oncological indications and multiple chemically distinct YAP/TEAD inhibitors are rapidly progressing towards clinical stages. Here, we tested the liability for CYP induction of a diverse set of YAP/TEAD inhibitors with different modes of action and TEAD isoform selectivity profiles in monolayers and 3D spheroids of primary human hepatocytes (PHH). We found that YAP/TEAD inhibition resulted in broad induction of CYPs in 2D monolayers, whereas if at all only marginal induction was seen in spheroid culture. Comprehensive RNA-Seq indicated that YAP/TEAD signaling was increased in 2D culture compared to spheroids, which was paralleled by elevated activities of the interacting transcription factors LXR and ESRRA, likely at least in part due to altered mechanosensing. Inhibition of this YAP/TEAD hyperactivation resulted in an overall reduction of hepatocyte dedifferentiation marked by increased hepatic functionality, including CYPs. These results thus demonstrate that the observed induction is due to on-target effects of the compounds rather than direct activation of xenobiotic sensing nuclear receptors. Combined, the presented data link hepatocyte dedifferentiation to YAP/TEAD dysregulation, reveal a novel non-canonical pathway of CYP induction and highlight the advantage of organotypic 3D cultures to predict clinically relevant pharmacokinetic properties, particularly for atypical induction mechanisms.Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.