Brosseau S, Abreu P, Bouchez C, Charon L, Kieffer Y, Gentric G, Picant V, Veith I, Camonis J, Descroix S, Mechta-Grigoriou F, Parrini M C, Zalcman G
U830 INSERM "Cancer, Heterogenity, Instability, Plasticity", Team "Stress and Cancer", Institut Curie Research Centre, 26 rue d'Ulm, 75248 Cedex 05, Paris, France.
Medicine Faculty, Université Paris Cité, 26 rue Henri Henri Huchard, 75018, Paris, France.
Mol Cell Biochem. 2025 Jan;480(1):231-248. doi: 10.1007/s11010-024-04949-7. Epub 2024 Mar 1.
The Yes-associated protein (YAP) oncoprotein has been linked to both metastases and resistance to targeted therapy of lung cancer cells. We aimed to investigate the effect of YAP pharmacological inhibition, using YAP/TEA domain (TEAD) transcription factor interaction inhibitors in chemo-resistant lung cancer cells. YAP subcellular localization, as a readout for YAP activation, cell migration, and TEAD transcription factor functional transcriptional activity were investigated in cancer cell lines with up-regulated YAP, with and without YAP/TEAD interaction inhibitors. Parental (A549) and paclitaxel-resistant (A549R) cell transcriptomes were analyzed. The half-maximal inhibitory concentration (IC) of paclitaxel or trametinib, which are Mitogen-Activated protein kinase and Erk Kinase (MEK) inhibitors, combined with a YAP/TEAD inhibitor (IV#6), was determined. A three-dimensional (3D) microfluidic culture device enabled us to study the effect of IV#6/paclitaxel combination on cancer cells isolated from fresh resected lung cancer samples. YAP activity was significantly higher in paclitaxel-resistant cell lines. The YAP/TEAD inhibitor induced a decreased YAP activity in A549, PC9, and H2052 cells, with reduced YAP nuclear staining. Wound healing assays upon YAP inhibition revealed impaired cell motility of lung cancer A549 and mesothelioma H2052 cells. Combining YAP pharmacological inhibition with trametinib in K-Ras mutated A549 cells recapitulated synthetic lethality, thereby sensitizing these cells to MEK inhibition. The YAP/TEAD inhibitor lowered the IC of paclitaxel in A549R cells. Differential transcriptomic analysis of parental and A549R cells revealed an increased YAP/TEAD transcriptomic signature in resistant cells, downregulated upon YAP inhibition. The YAP/TEAD inhibitor restored paclitaxel sensitivity of A549R cells cultured in a 3D microfluidic system, with lung cancer cells from a fresh tumor efficiently killed by YAP/TEAD inhibitor/paclitaxel doublet. Evidence of the YAP/TEAD transcriptional program's role in chemotherapy resistance paves the way for YAP therapeutic targeting.
Yes相关蛋白（YAP）癌蛋白与肺癌细胞的转移及对靶向治疗的耐药性均有关联。我们旨在研究使用YAP/TEA结构域（TEAD）转录因子相互作用抑制剂对化疗耐药肺癌细胞进行YAP药理学抑制的效果。在YAP上调的癌细胞系中，使用和不使用YAP/TEAD相互作用抑制剂，研究了YAP亚细胞定位（作为YAP激活的读数）、细胞迁移以及TEAD转录因子的功能性转录活性。分析了亲本（A549）和耐紫杉醇（A549R）细胞的转录组。测定了紫杉醇或曲美替尼（丝裂原活化蛋白激酶和细胞外信号调节激酶（MEK）抑制剂）与YAP/TEAD抑制剂（IV#6）联合使用时的半数最大抑制浓度（IC）。三维（3D）微流控培养装置使我们能够研究IV#6/紫杉醇组合对从新鲜切除的肺癌样本中分离出的癌细胞的作用。YAP活性在耐紫杉醇细胞系中显著更高。YAP/TEAD抑制剂在A549、PC9和H2052细胞中诱导YAP活性降低，YAP核染色减少。YAP抑制后的伤口愈合试验显示肺癌A549细胞和间皮瘤H2052细胞的细胞运动性受损。在K-Ras突变的A549细胞中将YAP药理学抑制与曲美替尼联合使用重现了合成致死性，从而使这些细胞对MEK抑制敏感。YAP/TEAD抑制剂降低了A549R细胞中紫杉醇的IC。亲本细胞和A549R细胞的差异转录组分析显示，耐药细胞中YAP/TEAD转录组特征增加，YAP抑制后下调。YAP/TEAD抑制剂恢复了在3D微流控系统中培养的A549R细胞对紫杉醇的敏感性，来自新鲜肿瘤的肺癌细胞被YAP/TEAD抑制剂/紫杉醇双联组合有效杀死。YAP/TEAD转录程序在化疗耐药中作用的证据为YAP的治疗靶向铺平了道路。
