肝细胞癌（HCC）是全球癌症相关死亡的主要原因。β-连环素（CTNNB1）突变型HCC占疾病30％的案例，但无精准治疗方法可用。我们使用从临床多激酶抑制剂（KI）中衍生的化学文库，对HCC器官oid进行了筛选，以鉴定WNTinib，这是一种在CTNNB1突变的人和小鼠模型中具有极高选择性的KI，包括患者样本。结合多组学和靶点参与分析，以及救治实验和体外和体内疗效研究，我们发现WNTinib优于临床KI，并在多个节点上抑制KIT/丝裂原活化蛋白激酶（MAPK）信号传导。此外，我们证明了相对于几种多重KI，WNTinib对BRAF和p38α激酶的结合减少是必要的，以避免补偿性反馈信号传导，通过EZH2转录抑制因子的核转位提供持久且选择性的突变β-连环素/Wnt靶点的转录抑制作用。我们的研究揭示了一种以前未知的机制，利用KIT/MAPK/EZH2途径强力并且选择性地拮抗CTNNB1突变型HCC，具有前所未有的广泛治疗指数。© 2023. 作者，独家授权Springer Nature America，Inc.

Hepatocellular carcinoma (HCC) is a leading cause of cancer-related deaths worldwide. β-Catenin (CTNNB1)-mutated HCC represents 30% of cases of the disease with no precision therapeutics available. Using chemical libraries derived from clinical multi-kinase inhibitor (KI) scaffolds, we screened HCC organoids to identify WNTinib, a KI with exquisite selectivity in CTNNB1-mutated human and murine models, including patient samples. Multiomic and target engagement analyses, combined with rescue experiments and in vitro and in vivo efficacy studies, revealed that WNTinib is superior to clinical KIs and inhibits KIT/mitogen-activated protein kinase (MAPK) signaling at multiple nodes. Moreover, we demonstrate that reduced engagement on BRAF and p38α kinases by WNTinib relative to several multi-KIs is necessary to avoid compensatory feedback signaling-providing a durable and selective transcriptional repression of mutant β-catenin/Wnt targets through nuclear translocation of the EZH2 transcriptional repressor. Our studies uncover a previously unknown mechanism to harness the KIT/MAPK/EZH2 pathway to potently and selectively antagonize CTNNB1-mutant HCC with an unprecedented wide therapeutic index.© 2023. The Author(s), under exclusive licence to Springer Nature America, Inc.