Functional RNAi Screens Define Distinct Protein Kinase Vulnerabilities in EGFR-Dependent HNSCC Cell Lines
Functional RNAi Screens Define Distinct Protein Kinase Vulnerabilities in EGFR-Dependent HNSCC Cell Lines
复制标题
DOI：
10.1124/mol.119.117804
复制
发表时间：
2019-12-01
影响因子：
3.6
通讯作者：
Heasley, Lynn E.
中科院分区：
文献类型：
--
作者：
Hinz, Trista K.;Kleczko, Emily K.;Heasley, Lynn E.
抑制性表皮生长因子受体（EGFR）抗体，西妥昔单抗，是一种批准的治疗头颈部鳞状细胞癌（HNSCC）。尽管在一些HNSCC患者中观察到肿瘤反应，但西妥昔单抗单独或与放疗或化疗联合不能产生长期控制或治愈。我们假设，一个灵活的受体酪氨酸激酶共激活信号网络支持HNSCC的生存在EGFR阻断的设置，并破坏该网络的药物将提供上级肿瘤控制时，与EGFR抑制剂组合。在这项工作中，我们提交了EGFR依赖性HNSCC细胞系的RNA干扰为基础的功能基因组学筛选，以确定，在一个公正的方式，生长和生存的必需蛋白激酶，以及合成的致命目标与EGFR拮抗剂联合抑制。雷帕霉素激酶（MTOR）和成红细胞增多症癌基因B（ERBB）3的机制靶标被鉴定为HNSCC细胞系中的高级必需激酶命中。通过不同的短发夹RNA（shRNA）和细胞系对AZD 8055的高敏感性证实了MTOR依赖性，而通过shRNA介导的沉默验证了ERBB 3依赖性。此外，使用泛ERBB抑制剂AZD 8931进行的合成致死激酶组shRNA筛选鉴定了细胞外信号调节激酶（ERK）促分裂原活化蛋白激酶途径的多种组分，与EGFR抑制剂单药治疗后ERK再活化和/或ERK途径不完全抑制一致。作为验证，不同的丝裂原活化蛋白激酶激酶（MEK）抑制剂与EGFR抑制剂吉非替尼和AZD 8931联合使用时产生协同生长抑制作用。研究结果确定ERBB 3和MTOR作为重要的药理学弱点在HNSCC和支持MEK和EGFR抑制剂相结合，以提高临床疗效HNSCC.SIGNIFICANCE声明许多癌症是由nonmutated受体酪氨酸激酶共激活网络，无视完全抑制与单一靶向药物驱动。本研究确定成红细胞增多症癌基因B（ERBB）3是表皮生长因子受体依赖性头颈部鳞状细胞癌（HNSCC）细胞系中的一种必需蛋白激酶，并与细胞外信号调节激酶丝裂原活化蛋白激酶途径发生合成致死性相互作用，为将泛ERBB和丝裂原活化蛋白激酶抑制剂联合用作HNSCC亚群的治疗方法提供了理论基础。
The inhibitory epidermal growth factor receptor (EGFR) antibody, cetuximab, is an approved therapy for head and neck squamous cell carcinoma (HNSCC). Despite tumor response observed in some HNSCC patients, cetuximab alone or combined with radio- or chemotherapy fails to yield long-term control or cures. We hypothesize that a flexible receptor tyrosine kinase coactivation signaling network supports HNSCC survival in the setting of EGFR blockade, and that drugs disrupting this network will provide superior tumor control when combined with EGFR inhibitors. In this work, we submitted EGFR-dependent HNSCC cell lines to RNA interference-based functional genomics screens to identify, in an unbiased fashion, essential protein kinases for growth and survival as well as synthetic lethal targets for combined inhibition with EGFR antagonists. Mechanistic target of rapamycin kinase (MTOR) and erythroblastosis oncogene B (ERBB)3 were identified as high-ranking essential kinase hits in the HNSCC cell lines. MTOR dependency was confirmed by distinct short hairpin RNAs (shRNAs) and high sensitivity of the cell lines to AZD8055, whereas ERBB3 dependency was validated by shRNA-mediated silencing. Furthermore, a synthetic lethal kinome shRNA screen with a pan-ERBB inhibitor, AZD8931, identified multiple components of the extracellular signal-regulated kinase (ERK) mitogen-activated protein kinase pathway, consistent with ERK reactivation and/or incomplete ERK pathway inhibition in response to EGFR inhibitor monotherapy. As validation, distinct mitogen-activated protein kinase kinase (MEK) inhibitors yielded synergistic growth inhibition when combined with the EGFR inhibitors, gefitinib and AZD8931. The findings identify ERBB3 and MTOR as important pharmacological vulnerabilities in HNSCC and support combining MEK and EGFR inhibitors to enhance clinical efficacy in HNSCC.SIGNIFICANCE STATEMENTMany cancers are driven by nonmutated receptor tyrosine kinase coactivation networks that defy full inhibition with single targeted drugs. This study identifies erythroblastosis oncogene B (ERBB) 3 as an essential protein kinase in epidermal growth factor receptor-dependent head and neck squamous cell cancer (HNSCC) cell lines and a synthetic lethal interaction with the extracellular signal-regulated kinase mitogen-activated protein kinase pathway that provides a rationale for combining pan-ERBB and mitogen-activated protein kinase inhibitors as a therapeutic approach in subsets of HNSCC.