Targeting USP9X-AMPK Axis in ARID1A-Deficient Hepatocellular Carcinoma.
Targeting USP9X-AMPK Axis in ARID1A-Deficient Hepatocellular Carcinoma.
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DOI：
10.1016/j.jcmgh.2022.03.009
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发表时间：
2022
影响因子：
7.2
通讯作者：
Li, Jing-Jing
中科院分区：
文献类型：
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作者：
Zhang, Feng-Kun;Ni, Qian-Zhi;Wang, Kang;Cao, Hui-Jun;Guan, Dong-Xian;Zhang, Er-Bin;Ma, Ning;Wang, Yi-Kang;Zheng, Qian-Wen;Xu, Sheng;Zhu, Bing;Chen, Tian-Wei;Xia, Ji;Qiu, Xiao-Song;Ding, Xu-Fen;Jiang, Hao;Qiu, Lin;Wang, Xiang;Chen, Wei;Cheng, Shu-Qun;Xie, Dong;Li, Jing-Jing
肝细胞癌是一种高度异质性的实体瘤，发病率和死亡率都很高。富含AT的相互作用域1A(ARID1A)在肝癌的突变中所占比例高达10%，然而，它在肝癌中的作用仍然存在争议，目前还没有建立针对性的治疗方法。用Western印迹和免疫组织化学方法检测ARID1a在临床标本中的表达。用聚集的规则间隔短回文重复序列(CRISPR)/CRISPR相关蛋白9(Cas9)基因敲除肝癌细胞株中的ARID1A，检测缺糖对细胞存活、增殖和凋亡的影响。用质谱分析寻找ARID1a相互作用的蛋白质，并通过免疫共沉淀和谷胱甘肽S转移酶下拉实验验证了结果。通过染色质免疫沉淀、谷胱甘肽S转移酶下拉、荧光素酶报告实验等方法研究ARID1A靶基因USP9X的调控。最后，进行药物治疗，以探索针对ARID1A缺陷型肝癌的药物在体内外的治疗潜力。我们的研究表明，ARID1A丢失对葡萄糖剥夺诱导的细胞死亡具有保护作用。机制研究表明，AIRD1a通过其C末端区域DUF3518将组蛋白脱乙酰酶1募集到USP9X启动子上，导致USP9X及其靶蛋白激酶AMP激活的催化亚基α2(PRKAA2)表达下调。在肝癌中，ARID1a基因敲除和1989年的∗截断突变体取消了这一效应，增加了USP9X启动子上H3K9和H3K27的乙酰化水平，上调了USP9X和蛋白激酶AMP激活的催化亚基α2的表达，从而介导了肿瘤细胞对葡萄糖饥饿的适应。化合物C能显著抑制ARID1A缺陷肿瘤的生长，延长荷瘤小鼠的存活时间。具有ARID1A突变的肝细胞癌患者可能受益于针对泛素特异性多肽酶9X-连锁(USP9X)-腺苷5‘-单磷酸激活蛋白激酶(AMPK)轴的综合致命治疗。
Hepatocellular carcinoma (HCC) is a highly heterogeneous solid tumor with high morbidity and mortality. AT-rich interaction domain 1A (ARID1A) accounts for up to 10% of mutations in liver cancer, however, its role in HCC remains controversial, and no targeted therapy has been established. The expression of ARID1A in clinical samples was examined by Western blot and immunohistochemical staining. ARID1A was knocked out by Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) in HCC cell lines, and the effects of glucose deprivation on cell viability, proliferation, and apoptosis were measured. Mass spectrometry analysis was used to find ARID1A-interacting proteins, and the result was verified by co-immunoprecipitation and Glutathione S Transferase (GST) pull-down. The regulation of ARID1A target gene USP9X was investigated by chromatin immunoprecipitation, Glutathione S Transferase (GST) pull-down, luciferase reporter assay, and so forth. Finally, drug treatments were performed to explore the therapeutic potential of the agents targeting ARID1A-deficient HCC in vitro and in vivo. Our study has shown that ARID1A loss protected cells from glucose deprivation–induced cell death. A mechanism study disclosed that AIRD1A recruited histone deacetylase 1 via its C-terminal region DUF3518 to the promoter of USP9X, resulting in down-regulation of USP9X and its target protein kinase AMP-activated catalytic subunit α2 (PRKAA2). ARID1A knockout and a 1989∗ truncation mutant in HCC abolished this effect, increased the levels of H3K9 and H3K27 acetylation at the USP9X promoter, and up-regulated the expression of USP9X and protein kinase AMP-activated catalytic subunit α2 (PRKAA2), which mediated the adaptation of tumor cells to glucose starvation. Compound C dramatically inhibited the growth of ARID1A-deficient tumors and prolongs the survival of tumor-bearing mice. HCC patients with ARID1A mutation may benefit from synthetic lethal therapy targeting the ubiquitin-specific peptidase 9 X-linked (USP9X)–adenosine 5‘-monophosphate–activated protein kinase (AMPK) axis.
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影响因子：
13.3
作者：
Liu H;Ding J;Köhnlein K;Urban N;Ori A;Villavicencio-Lorini P;Walentek P;Klotz LO;Hollemann T;Pfirrmann T
通讯作者：
Pfirrmann T
影响因子：
3.7
作者：
Gupta, Piyushi;Singh, Ankita;Sen, Ellora
通讯作者：
Sen, Ellora
影响因子：
6.4
作者：
Ali, Jennifer L.;Lagasse, Brittany J.;Nachtigal, Mark W.
通讯作者：
Nachtigal, Mark W.
影响因子：
13.3
作者：
Driessen, Stefan;Berleth, Niklas;Stork, Bjoern
通讯作者：
Stork, Bjoern
影响因子：
82.9
作者：
通讯作者：
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