Detection of phenotype-specific therapeutic vulnerabilities in breast cells using a CRISPR loss-of-function screen.
Detection of phenotype-specific therapeutic vulnerabilities in breast cells using a CRISPR loss-of-function screen.
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DOI：
10.1002/1878-0261.12951
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发表时间：
2021-08
影响因子：
6.6
通讯作者：
Prasmickaite L
中科院分区：
文献类型：
--
作者：
Barkovskaya A;Goodwin CM;Seip K;Hilmarsdottir B;Pettersen S;Stalnecker C;Engebraaten O;Briem E;Der CJ;Moestue SA;Gudjonsson T;Maelandsmo GM;Prasmickaite L
上皮状态和间质状态之间的细胞表型可塑性与癌症治疗的转移和异质反应有关，并且仍然是三阴性乳腺癌（TNBC）治疗的挑战。在这里，我们使用同基因人乳腺上皮细胞系 D492 和 D492M，分别代表上皮和间质表型。我们采用针对 2240 个基因“可药物基因组”的 CRISPR-Cas9 功能丧失筛选来识别表型特异性漏洞。具有上皮表型的细胞更容易丢失与 EGFR-RAS-MAPK 信号传导相关的基因，而间充质样细胞对 G2-M 细胞周期调节因子的敲除更加敏感。此外，我们还发现了以表型特异性方式对 mTOR 抑制剂依维莫司和化疗药物氟尿嘧啶敏感的基因敲除。具体来说，EGFR 和脂肪酸合酶 (FASN) 的缺失分别增加了药物在上皮和间质表型中的有效性。这些与表型相关的遗传脆弱性已通过 EGFR（吉非替尼）、G2-M 转变（STLC）和 FASN（Fasnall）的靶向抑制剂得到证实。总之，CRISPR-Cas9 功能丧失筛选能够识别表型特异性遗传漏洞，从而确定可行的靶点和有希望的治疗组合。表型异质性阻碍了抗癌治疗的功效。在这里，我们在显示相反 EMT 表型的同基因乳腺细胞系中应用了 CRISPR-Cas9 功能丧失筛选的“可药物基因组”。我们确定了表型特异性遗传脆弱性（单独或在“治疗压力”下），它们代表不同表型中的可操作靶点：上皮表型中的 EGFR-MAPK 信号传导，以及间充质表型细胞中的 G2-M 转变和 FASN。
Cellular phenotype plasticity between the epithelial and mesenchymal states has been linked to metastasis and heterogeneous responses to cancer therapy, and remains a challenge for the treatment of triple‐negative breast cancer (TNBC). Here, we used isogenic human breast epithelial cell lines, D492 and D492M, representing the epithelial and mesenchymal phenotypes, respectively. We employed a CRISPR‐Cas9 loss‐of‐function screen targeting a 2240‐gene ‘druggable genome’ to identify phenotype‐specific vulnerabilities. Cells with the epithelial phenotype were more vulnerable to the loss of genes related to EGFR‐RAS‐MAPK signaling, while the mesenchymal‐like cells had increased sensitivity to knockout of G2‐M cell cycle regulators. Furthermore, we discovered knockouts that sensitize to the mTOR inhibitor everolimus and the chemotherapeutic drug fluorouracil in a phenotype‐specific manner. Specifically, loss of EGFR and fatty acid synthase (FASN) increased the effectiveness of the drugs in the epithelial and mesenchymal phenotypes, respectively. These phenotype‐associated genetic vulnerabilities were confirmed using targeted inhibitors of EGFR (gefitinib), G2‐M transition (STLC), and FASN (Fasnall). In conclusion, a CRISPR‐Cas9 loss‐of‐function screen enables the identification of phenotype‐specific genetic vulnerabilities that can pinpoint actionable targets and promising therapeutic combinations. Phenotypic heterogeneity hampers anticancer treatment efficacy. Here, we applied a CRISPR‐Cas9 loss‐of‐function screen of a ‘druggable genome’ in isogenic breast cell lines displaying opposing EMT phenotypes. We identified phenotype‐specific genetic vulnerabilities (alone or under ‘therapy pressure’) that represent actionable targets in distinct phenotypes: EGFR‐MAPK signaling in the epithelial phenotype, and G2‐M transition and FASN in the mesenchymal phenotype cells.
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影响因子：
10.5
作者：
Gudjonsson, T;Villadsen, R;Petersen, OW
通讯作者：
Petersen, OW
影响因子：
8
作者：
Ingthorsson S;Andersen K;Hilmarsdottir B;Maelandsmo GM;Magnusson MK;Gudjonsson T
通讯作者：
Gudjonsson T
影响因子：
28.4
作者：
Falchook, Gerald;Coleman, Robert L.;Schilder, Russell J.
通讯作者：
Schilder, Russell J.
影响因子：
64.8
作者：
Fischer KR;Durrans A;Lee S;Sheng J;Li F;Wong ST;Choi H;El Rayes T;Ryu S;Troeger J;Schwabe RF;Vahdat LT;Altorki NK;Mittal V;Gao D
通讯作者：
Gao D
影响因子：
45.3
作者：
Denduluri, Neelima;Chavez-MacGregor, Mariana;Giordano, Sharon H.
通讯作者：
Giordano, Sharon H.