N6-Methyladenosine-Mediated Up-Regulation of FZD10 Regulates Liver Cancer Stem Cells' Properties and Lenvatinib Resistance Through WNT/b-Catenin and Hippo Signaling Pathways
N6-Methyladenosine-Mediated Up-Regulation of FZD10 Regulates Liver Cancer Stem Cells' Properties and Lenvatinib Resistance Through WNT/b-Catenin and Hippo Signaling Pathways
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DOI：
10.1053/j.gastro.2023.01.041
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发表时间：
2023-04-20
期刊：
影响因子：
29.4
通讯作者：
Xiang, Daimin
中科院分区：
文献类型：
--
作者：
Wang, Jinghan;Yu, Hongming;Xiang, Daimin
背景与目的：肝细胞癌（HCC）是全球癌症相关死亡的主要原因之一，但缺乏早期诊断生物标志物和治疗靶点。耐药性是大多数hcc相关死亡的原因，但对耐药性的潜在机制仍知之甚少。方法：采用RNA测序方法鉴定Frizzled -10 （FZD10）在肝癌干细胞（CSCs）中的表达，并采用实时聚合酶链反应和免疫组织化学方法进行验证。体外和体内实验探讨FZD10对肝CSC扩张和lenvatinib耐药的影响。采用RNA测序、RNA结合蛋白免疫沉淀和荧光素酶报告分析来探索fzd10介导的肝CSCs扩增和lenvinib耐药的机制。结果：肝脏CSCs中FZD10的激活是通过mettl3依赖性n6 -甲基腺苷甲基化FZD10信使RNA介导的。功能研究显示FZD10通过激活b-连环蛋白和YAP1促进肝CSCs的自我更新、致瘤性和转移。FZD10-b-catenin/YAP1轴在肝CSCs中被激活并预测不良预后。此外，FZD10-b-catenin/c-Jun轴转录激活METTL3表达，形成一个正反馈回路。重要的是，FZD10/b-catenin/c-Jun/ MEK/ERK轴决定了肝癌细胞对lenvatinib治疗的反应。对患者队列、患者来源的肿瘤类器官和患者来源的异种移植物的分析进一步表明，FZD10可能预测lenvatinib在HCC患者中的临床获益。此外，用针对FZD10或b-catenin抑制剂的腺相关病毒治疗lenvatinib耐药HCC可恢复lenvatinib应答。结论：FZD10表达升高可促进肝CSCs的扩张和lenvatinib耐药，表明FZD10表达是人类HCC的一种新的预后生物标志物和治疗靶点。
BACKGROUND & AIMS: Hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related deaths worldwide, but there is a deficiency of early diagnosis biomarkers and thera-peutic targets. Drug resistance accounts for most HCC-related deaths, yet the mechanisms underlying drug resistance remain poorly understood. METHODS: Expression of Frizzled -10 (FZD10) in liver cancer stem cells (CSCs) was identified by means of RNA sequencing and validated by means of real-time polymerase chain reaction and immunohistochemistry. In vitro and in vivo experiments were used to assess the effect of FZD10 on liver CSC expansion and lenvatinib resistance. RNA sequencing, RNA binding protein immunoprecipitation, and luciferase report assays were applied to explore the mechanism underlying FZD10-mediated liver CSCs expansion and lenvati-nib resistance. RESULTS: Activation of FZD10 in liver CSCs was mediated by METTL3-dependent N6-methyladenosine methyl-ation of FZD10 messenger RNA. Functional studies revealed that FZD10 promotes self-renewal, tumorigenicity, and metas-tasis of liver CSCs via activating b-catenin and YAP1. The FZD10-b-catenin/YAP1 axis is activated in liver CSCs and predicts poor prognosis. Moreover, FZD10-b-catenin/c-Jun axis transcriptionally activates METTL3 expression, forming a pos-itive feedback loop. Importantly, the FZD10/b-catenin/c-Jun/ MEK/ERK axis determines the responses of hepatoma cells to lenvatinib treatment. Analysis of patient cohort, patient-derived tumor organoids, and patient-derived xenografts further suggest that FZD10 might predict lenvatinib clinical benefit in patients with HCC. Furthermore, treatment of lenvatinib-resistant HCC with adeno-associated virus targeting FZD10 or a b-catenin inhibitor restored lenvatinib response. CONCLUSIONS: Elevated FZD10 expression promotes expan-sion of liver CSCs and lenvatinib resistance, indicating that FZD10 expression is a novel prognostic biomarker and thera-peutic target for human HCC.