A genome-wide CRISPR/Cas9 screen in acute myeloid leukemia cells identifies regulators of TAK-243 sensitivity
A genome-wide CRISPR/Cas9 screen in acute myeloid leukemia cells identifies regulators of TAK-243 sensitivity
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DOI：
10.1172/jci.insight.141518
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发表时间：
2021-03-08
期刊：
影响因子：
8
通讯作者：
Schimmer, Aaron D.
中科院分区：
文献类型：
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作者：
Barghout, Samir H.;Aman, Ahmed;Schimmer, Aaron D.
TAK-243是一类泛素样修饰物激活酶1的抑制剂，它催化泛素活化，这是泛素化级联的第一步。基于其临床前疗效和耐受性，TAK-243已进入晚期恶性肿瘤的I期临床试验。尽管如此，TAK-243敏感性的决定因素在很大程度上仍然未知。在这里，我们在存在TAK-243的急性髓性白血病（AML）细胞中进行了全基因组CRISPR/Cas9敲除筛选，以鉴定TAK-243作用所必需的基因。我们发现BEN结构域蛋白3 （BEN domain containing protein 3, BEND3）是一种转录抑制因子和染色质组织的调节因子，是在体外和体内对TAK-243具有抗性的顶级基因。敲除BEND3可抑制TAK-243对泛素化、蛋白毒性应激和DNA损伤反应的影响。BEND3敲除上调atp结合盒外排转运体乳腺癌耐药蛋白（BCRP; ABCG2），降低细胞内TAK-243水平。TAK-243敏感性与BCRP在不同来源癌细胞系中的表达相关。此外，BCRP的化学抑制和基因敲低使内在耐药的高BCRP细胞对TAK-243敏感。因此，我们的数据表明BEND3调节BCRP的表达，而TAK-243是BCRP的底物。此外，BCRP表达可作为TAK-243敏感性的预测因子。
TAK-243 is a first-in-class inhibitor of ubiquitin-like modifier activating enzyme 1 that catalyzes ubiquitin activation, the first step in the ubiquitylation cascade. Based on its preclinical efficacy and tolerability, TAK-243 has been advanced to phase I clinical trials in advanced malignancies. Nonetheless, the determinants of TAK-243 sensitivity remain largely unknown. Here, we conducted a genome-wide CRISPR/Cas9 knockout screen in acute myeloid leukemia (AML) cells in the presence of TAK-243 to identify genes essential for TAK-243 action. We identified BEN domain-containing protein 3 (BEND3), a transcriptional repressor and a regulator of chromatin organization, as the top gene whose knockout confers resistance to TAK-243 in vitro and in vivo. Knockout of BEND3 dampened TAK-243 effects on ubiquitylation, proteotoxic stress, and DNA damage response. BEND3 knockout upregulated the ATP-binding cassette efflux transporter breast cancer resistance protein (BCRP; ABCG2) and reduced the intracellular levels of TAK-243. TAK-243 sensitivity correlated with BCRP expression in cancer cell lines of different origins. Moreover, chemical inhibition and genetic knockdown of BCRP sensitized intrinsically resistant high-BCRP cells to TAK-243. Thus, our data demonstrate that BEND3 regulates the expression of BCRP for which TAK-243 is a substrate. Moreover, BCRP expression could serve as a predictor of TAK-243 sensitivity.