CDK12 inhibition mediates DNA damage and is synergistic with sorafenib treatment in hepatocellular carcinoma
CDK12 inhibition mediates DNA damage and is synergistic with sorafenib treatment in hepatocellular carcinoma
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CDK12 抑制介导 DNA 损伤，并与索拉非尼治疗肝细胞癌具有协同作用
DOI：
10.1136/gutjnl-2019-318506
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发表时间：
2020-04-01
期刊：
影响因子：
24.5
通讯作者：
Bernards, Rene
中科院分区：
文献类型：
--
作者：
Wang, Cun;Wang, Hui;Bernards, Rene
肝细胞癌（HCC）是世界范围内最常见的恶性肿瘤之一，也是癌症相关死亡的主要原因。几种治疗选择，如索拉非尼和瑞格非尼，仅为HCC患者提供适度的生存益处。本研究的目的是为HCC患者确定新的可药物候选基因。设计进行靶向所有已知人类激酶的无偏倚CRISPR（成簇的规则间隔短回文重复序列）功能丧失遗传筛选以鉴定HCC细胞的脆弱性。通过全转录组测序（RNA-Seq）和生物信息学分析，探讨细胞周期蛋白依赖性激酶12（CDK 12）抑制剂在肝癌细胞中的作用机制。使用多种体外和体内测定来研究CDK 12抑制和索拉非尼的组合的协同效应。结果我们鉴定出CDK 12是大多数肝癌细胞系所必需的。使用短发夹RNA（shRNA）抑制CDK 12或通过共价小分子抑制剂THZ 531抑制CDK 12导致稳健的增殖抑制。THZ 531优先抑制DNA修复相关基因的表达，并诱导HCC细胞系中强烈的DNA损伤反应。THZ 531和索拉非尼的组合通过诱导HCC细胞中的凋亡或衰老显示出惊人的协同作用。THZ 531和索拉非尼之间的协同作用可能源自THZ 531损害由索拉非尼治疗诱导的HCC细胞的适应性应答的概念。结论CDK 12有可能成为治疗肝癌的药物靶点。THZ 531和索拉非尼的惊人协同作用表明了这种难以治疗的癌症的潜在联合疗法。
Objectives Hepatocellular carcinoma (HCC) is one of the most frequent malignancies and a major leading cause of cancer-related deaths worldwide. Several therapeutic options like sorafenib and regorafenib provide only modest survival benefit to patients with HCC. This study aims to identify novel druggable candidate genes for patients with HCC. Design A non-biased CRISPR (clustered regularly interspaced short palindromic repeats) loss-of-function genetic screen targeting all known human kinases was performed to identify vulnerabilities of HCC cells. Whole-transcriptome sequencing (RNA-Seq) and bioinformatics analyses were performed to explore the mechanisms of the action of a cyclin-dependent kinase 12 (CDK12) inhibitor in HCC cells. Multiple in vitro and in vivo assays were used to study the synergistic effects of the combination of CDK12 inhibition and sorafenib. Results We identify CDK12 as critically required for most HCC cell lines. Suppression of CDK12 using short hairpin RNAs (shRNAs) or its inhibition by the covalent small molecule inhibitor THZ531 leads to robust proliferation inhibition. THZ531 preferentially suppresses the expression of DNA repair-related genes and induces strong DNA damage response in HCC cell lines. The combination of THZ531 and sorafenib shows striking synergy by inducing apoptosis or senescence in HCC cells. The synergy between THZ531 and sorafenib may derive from the notion that THZ531 impairs the adaptive responses of HCC cells induced by sorafenib treatment. Conclusion Our data highlight the potential of CDK12 as a drug target for patients with HCC. The striking synergy of THZ531 and sorafenib suggests a potential combination therapy for this difficult to treat cancer.