Enhancing efficacy of the MEK inhibitor trametinib with paclitaxel in KRAS-mutated colorectal cancer

Susmita Ghosh, Fan Fan, Reid T Powell, Yong Park, Clifford C Stephan, Scott Kopetz, Lee Ellis, Rajat Bhattacharya
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Abstract

Background: KRAS is frequently mutated in the tumors of patients with metastatic colorectal cancer (mCRC) and thus represents a valid target for therapy. However, the strategies of targeting KRAS directly and targeting the downstream effector mitogen-activated protein kinase kinase (MEK) via monotherapies have shown limited efficacy. Thus, there is a strong need for novel, effective combination therapies to improve MEK-inhibitor efficacy in patients with KRAS-mutated mCRC. Objective: Our objective was to identify novel drug combinations that enhance MEK-inhibitor efficacy in patients with KRAS-mutated mCRC. Design: In this study, we performed unbiased high-throughput screening (HTS) to identify drugs that enhance the efficacy of MEK inhibitors in vitro, and we validated the efficacy of the drugs in vivo. Methods: HTS was performed using 3-dimensional CRC spheroids. Trametinib, the anchor drug, was probed with 2 clinically ready libraries of 252 drugs to identify effective drug combinations. The effects of the drug combinations on CRC cell proliferation and apoptosis were further validated using cell growth assays, flow cytometry, and biochemical assays. Proteomic and immunostaining studies were performed to determine the effects of the drugs on molecular signaling and cell division. The effects of the drug combinations were examined in vivo using CRC patient-derived xenografts. Results: HTS identified paclitaxel as being synergistic with trametinib. In vitro validation showed that, compared with monotherapies, this drug combination demonstrated strong inhibition of cell growth, reduced colony formation, and enhanced apoptosis in multiple KRAS-mutated CRC cell lines. Mechanistically, combining trametinib with paclitaxel led to alterations in signaling mediators that block cell cycle progression and increases in microtubule stability that resulted in significantly higher defects in the mitosis. Finally, the combination of trametinib with paclitaxel exhibited significant inhibition of tumor growth in several KRAS-mutant patient-derived xenograft mouse models. Conclusion: Our data provide evidence supporting clinical trials of trametinib with paclitaxel as a novel therapeutic option for patients with KRAS-mutated, metastatic CRC.
在 KRAS 突变的结直肠癌中增强 MEK 抑制剂曲美替尼与紫杉醇的疗效
背景:在转移性结直肠癌(mCRC)患者的肿瘤中,KRAS经常发生突变,因此是一个有效的治疗靶点。然而,直接靶向 KRAS 和通过单一疗法靶向下游效应物质丝裂原活化蛋白激酶激酶 (MEK) 的策略疗效有限。因此,KRAS突变的mCRC患者亟需新型、有效的联合疗法来提高MEK抑制剂的疗效。目标:我们的目标是找出能提高 KRAS 突变 mCRC 患者 MEK 抑制剂疗效的新型药物组合。设计:在本研究中,我们进行了无偏见的高通量筛选(HTS),以确定能在体外增强 MEK 抑制剂疗效的药物,并在体内验证了这些药物的疗效。方法:使用三维 CRC 球形体进行高通量筛选。将锚定药物曲美替尼与 2 个包含 252 种药物的临床可用药物库进行比对,以确定有效的药物组合。利用细胞生长测定、流式细胞仪和生化测定进一步验证了药物组合对 CRC 细胞增殖和凋亡的影响。还进行了蛋白质组学和免疫染色研究,以确定药物对分子信号转导和细胞分裂的影响。使用源自 CRC 患者的异种移植体在体内检测了药物组合的效果。结果HTS确定紫杉醇与曲美替尼具有协同作用。体外验证显示,与单一疗法相比,这种药物组合在多种 KRAS 突变的 CRC 细胞系中表现出强烈的抑制细胞生长、减少集落形成和增强细胞凋亡的作用。从机理上讲,曲美替尼与紫杉醇联用会导致阻碍细胞周期进展的信号介质发生改变,微管稳定性增加,从而导致有丝分裂缺陷显著增加。最后,曲美替尼与紫杉醇的联合用药在多个 KRAS 突变患者异种移植小鼠模型中显示出显著的抑制肿瘤生长作用。结论我们的数据提供了支持曲美替尼联合紫杉醇临床试验的证据,可作为KRAS突变转移性CRC患者的一种新型治疗选择。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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