Synergistic activity of mTORC1/2 kinase and MEK inhibitors suppresses pediatric low-grade glioma tumorigenicity and vascularity
Synergistic activity of mTORC1/2 kinase and MEK inhibitors suppresses pediatric low-grade glioma tumorigenicity and vascularity
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DOI：
10.1093/neuonc/noz230
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发表时间：
2020-04-01
期刊：
影响因子：
15.9
通讯作者：
Raabe, Eric H.
中科院分区：
文献类型：
--
作者：
Arnold, Antje;Yuan, Ming;Raabe, Eric H.
背景。儿童低级别胶质瘤（pLGG）是最常见的儿童脑肿瘤。许多患有不可切除或复发/难治性肿瘤的患者具有严重的终身残疾。大多数 pLGG 具有增加 Ras/丝裂原激活蛋白激酶 (MAPK) 途径活性的突变。哺乳动物雷帕霉素靶点 (mTOR) 的激活也是 pLGG 的一个标志。因此，我们假设雷帕霉素复合物 1 和 2 (TORC1/2) 激酶抑制剂 TAK228 的双重靶点将与 pLGG 中丝裂原激活的细胞外信号调节激酶 (MEK) 抑制剂曲美替尼 (Trametinib) 产生协同作用。我们在患者来源的 pLGG 细胞系中测试了 TAK228 和曲美替尼，该细胞系含有 pLGG 驱动因素，包括 BRAF(V600E) 和 1 型神经纤维瘤病缺失。我们测量了细胞增殖、途径抑制、细胞死亡和衰老。使用 Chou-Talalay 方法通过 MTS 测定分析协同作用。在体内，我们测试了总体存活率和通路抑制，并进行了增殖和血管化的免疫组织化学分析。我们进行了划痕实验并测量了人脐静脉内皮细胞 (HUVEC) 中血管生成蛋白的激活情况。结果。 TAK228 在所有测试的细胞系中以临床相关剂量与 pLGG 中的曲美替尼协同作用，抑制增殖，诱导细胞凋亡，并以细胞系依赖性方式引起衰老。与单一治疗和对照队列相比，联合治疗使中位生存期增加了 70%，并减少了肿瘤体积。根据 CD31 和 CD34 的测量，肿瘤的血管化减少。与单独使用每种药物相比，联合治疗可阻断 HUVEC 细胞中粘着斑激酶 (FAK) 和肉瘤原癌基因非受体酪氨酸激酶 (SRC) 的激活，并减少 HUVEC 迁移。结论。 TAK228 和曲美替尼的组合协同抑制 pLGG 的生长。这些药物协同作用，通过阻断 FAK 和 SRC 的激活来减少肿瘤血管分布以及内皮细胞的生长和迁移。
Background. Pediatric low-grade glioma (pLGG) is the most common childhood brain tumor. Many patients with unresectable or recurrent/refractory tumors have significant lifelong disability. The majority of pLGG have mutations increasing the activity of the Ras/mitogen-activated protein kinase (MAPK) pathway. Activation of mammalian target of rapamycin (mTOR) is also a hallmark of pLGG. We therefore hypothesized that the dual target of rapamycin complexes 1 and 2 (TORC1/2) kinase inhibitor TAK228 would synergize with the mitogen-activated extracellular signal-regulated kinase (MEK) inhibitor trametinib in pLGG.Methods. We tested TAK228 and trametinib in patient-derived pLGG cell lines harboring drivers of pLGG including BRAF(V600E) and neurofibromatosis type 1 loss. We measured cell proliferation, pathway inhibition, cell death, and senescence. Synergy was analyzed via MTS assay using the Chou-Talalay method. In vivo, we tested for overall survival and pathway inhibition and performed immunohistochemistry for proliferation and vascularization. We performed a scratch assay and measured angiogenesis protein activation in human umbilical vein endothelial cells (HUVECs).Results. TAK228 synergized with trametinib in pLGG at clinically relevant doses in all tested cell lines, suppressing proliferation, inducing apoptosis, and causing senescence in a cell line-dependent manner. Combination treatment increased median survival by 70% and reduced tumor volume compared with monotreatment and control cohorts. Vascularization of tumors decreased as measured by CD31 and CD34. Combination treatment blocked activation of focal adhesion kinase (FAK) and sarcoma proto-oncogene non-receptor tyrosine kinase (SRC) in HUVEC cells and reduced HUVEC migration compared with each drug alone.Conclusions. The combination of TAK228 and trametinib synergized to suppress the growth of pLGG. These agents synergized to reduce tumor vascularity and endothelial cell growth and migration by blocking activation of FAK and SRC.