科研成果: “Genome-wide CRISPR-Cas9 screen reveals selective vulnerability of ATRX-mutant cancers to WEE1 inhibition” (王晓月)

发布时间:2020-11-04


导读:  王晓月课题组于2020年2月1日在”Cancer Research”杂志发表研究论文“Genome-wide CRISPR-Cas9 screen reveals selective vulnerability of ATRX-mutant cancers to WEE1 inhibition”。该研究工作通过全基因组范围的CRISPR筛选,发现细胞周期检查点激酶WEE1是ATRX的合成致死基因并揭示了其合成致死机制。

文章链接:https://cancerres.aacrjournals.org/content/80/3/510

1. 全基因组范围CRISPR筛选鉴定ATRX合成致死基因

 

ATRX是SWI/SNF染色质重塑复合物的一员,肿瘤基因组研究表明ATRX基因在多种肿瘤中发生突变,包括胰腺神经内分泌肿瘤、神经胶质瘤、肝癌、神经母细胞瘤等。ATRX突变主要表现为无义突变,从而导致其蛋白功能缺失,表明ATRX在这些肿瘤中发挥抑癌基因的作用。

为了寻找ATRX突变型肿瘤的潜在治疗靶点,我们利用CRISPR/Cas9基因组编辑技术,进行了全基因组范围的合成致死筛选。在敲除ATRX基因的肝癌细胞系中,我们共鉴定出58个基因,其中包括细胞周期检查点激酶WEE1WEE1对ATRX缺失细胞的生长非常关键。WEE1抑制剂AZD1775对几种ATRX缺失型肝癌细胞系以及 PLC/PRF/5来源的移植瘤生长均有明显的抑制作用。进一步研究发现,对WEE1抑制剂敏感性的增加是由DNA损伤诱导的细胞凋亡积累所致。此外,AZD1775还选择性抑制具有ATRX突变的胶质瘤患者来源的原代细胞系的增殖,表明WEE1ATRX之间的合成致死关系可以在更广泛的肿瘤类型中得以利用。由于WEE1抑制剂已经在多个II期临床试验中进行了研究,我们的研究为针对ATRX缺失癌症的临床治疗提供了新的策略。相关工作发表在Cancer Research杂志,并被选为当期研究亮点。https://cancerres.aacrjournals.org/content/80/3/375

 

【摘要】The tumor suppressor gene ATRX is frequently mutated in a variety of tumors including gliomas and liver cancers, which are highly unresponsive to current therapies. Here, we performed a genome-wide synthetic lethal screen, using CRISPR-Cas9 genome editing, to identify potential therapeutic targets specific for ATRX-mutated cancers. In isogenic hepatocellular carcinoma (HCC) cell lines engineered for ATRX loss, we identified 58 genes, including the checkpoint kinase WEE1, uniquely required for the cell growth of ATRX null cells. Treatment with the WEE1 inhibitor AZD1775 robustly inhibited the growth of several ATRX-deficient HCC cell lines in vitro, as well as xenografts in vivo. The increased sensitivity to the WEE1 inhibitor was caused by accumulated DNA damage-induced apoptosis. AZD1775 also selectively inhibited the proliferation of patient-derived primary cell lines from gliomas with naturally occurring ATRX mutations, indicating that the synthetic lethal relationship between WEE1 and ATRX could be exploited in a broader spectrum of human tumors. As WEE1 inhibitors have been investigated in several phase II clinical trials, our discovery provides the basis for an easily clinically testable therapeutic strategy specific for cancers deficient in ATRX.

 

该研究由中国医学科学院基础医学研究所和医学分子生物学国家重点实验室王晓月团队与国家癌症中心焦宇辰团队共同完成。基础医学研究所和重点实验室梁俊波博士、国家癌症中心赵宏教授、杜克大学Bill H. Diplas和北京协和医院刘松博士为本文的共同第一作者,王晓月、焦宇辰为该论文的共同通讯作者。此研究获得中国医学科学院医学与健康科技创新工程基金(2017-I2M-4-003;2017-I2M-4-002)、国家自然科学基金(81472559;81502420)、医学分子生物学国家重点实验室自主研究课题(2060204)等资助。杜克大学医学中心阎海教授、中山大学肿瘤中心曾益新教授参与了该项研究。

医学分子生物学国家重点实验室