A high-throughput kinome screen reveals serum/glucocorticoid-regulated kinase 1 as a therapeutic target for NF2-deficient meningiomas.
A high-throughput kinome screen reveals serum/glucocorticoid-regulated kinase 1 as a therapeutic target for NF2-deficient meningiomas.
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DOI：
10.18632/oncotarget.4858
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发表时间：
2015-07-10
期刊：
影响因子：
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通讯作者：
Ramesh V
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文献类型：
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作者：
Beauchamp RL;James MF;DeSouza PA;Wagh V;Zhao WN;Jordan JT;Stemmer-Rachamimov A;Plotkin SR;Gusella JF;Haggarty SJ;Ramesh V
脑膜瘤是成人颅内最常见的原发性肿瘤。所有与神经纤维瘤病2（NF 2）相关的脑膜瘤和约60%的散发性脑膜瘤显示NF 2肿瘤抑制蛋白缺失。对于进行性和复发性脑膜瘤没有有效的药物治疗。我们之前的工作证明了mTORC 1信号的异常激活，导致正在进行的针对NF 2和散发性脑膜瘤患者的雷帕霉素类似物的临床试验。在这里，我们进行了高通量的激酶组筛选，以确定在NF 2缺陷型脑膜瘤细胞中负责mTORC 1通路激活的激酶。其中最热门的候选者是mTORC 2特异性靶血清/糖皮质激素调节激酶1（SGK 1）和p21激活激酶1（PAK 1）。在NF 2缺陷型脑膜瘤细胞中，抑制SGK 1可挽救mTORC 1激活，SGK 1激活对双重mTORC 1/2抑制剂AZD 2014敏感，但对雷帕霉素不敏感。PAK 1抑制也导致mTORC 1信号减弱，而不是mTORC 2信号，这表明mTORC 2/SGK 1和Rac 1/PAK 1通路独立负责NF 2缺陷型脑膜瘤中的mTORC 1激活。使用CRISPR-Cas9基因组编辑，我们生成了表达或缺乏NF 2的等基因人类蛛网膜细胞系（AC），这是脑膜瘤的起源细胞类型。NF 2缺失型CRISPR AC重现了NF 2缺陷型脑膜瘤细胞的信号传导。有趣的是，我们在NF 2-CRISPR AC和原发性NF 2阴性脑膜瘤细胞系中观察到SGK 1转录和蛋白表达增加。此外，我们证明了双重mTORC 1/mTORC 2抑制剂AZD 2014在阻断脑膜瘤细胞增殖方面上级雷帕霉素和PAK抑制剂FRAX 597。重要的是，AZD 2014目前正在几项癌症临床试验中使用。因此，我们认为AZD 2014可能为复发性和进展性脑膜瘤提供优于雷帕霉素类似物的治疗优势。
Meningiomas are the most common primary intracranial adult tumor. All Neurofibromatosis 2 (NF2)-associated meningiomas and ~60% of sporadic meningiomas show loss of NF2 tumor suppressor protein. There are no effective medical therapies for progressive and recurrent meningiomas. Our previous work demonstrated aberrant activation of mTORC1 signaling that led to ongoing clinical trials with rapamycin analogs for NF2 and sporadic meningioma patients. Here we performed a high-throughput kinome screen to identify kinases responsible for mTORC1 pathway activation in NF2-deficient meningioma cells. Among the emerging top candidates were the mTORC2-specific target serum/glucocorticoid-regulated kinase 1 (SGK1) and p21-activated kinase 1 (PAK1). In NF2-deficient meningioma cells, inhibition of SGK1 rescues mTORC1 activation, and SGK1 activation is sensitive to dual mTORC1/2 inhibitor AZD2014, but not to rapamycin. PAK1 inhibition also leads to attenuated mTORC1 but not mTORC2 signaling, suggesting that mTORC2/SGK1 and Rac1/PAK1 pathways are independently responsible for mTORC1 activation in NF2-deficient meningiomas. Using CRISPR-Cas9 genome editing, we generated isogenic human arachnoidal cell lines (ACs), the origin cell type for meningiomas, expressing or lacking NF2. NF2-null CRISPR ACs recapitulate the signaling of NF2-deficient meningioma cells. Interestingly, we observe increased SGK1 transcription and protein expression in NF2-CRISPR ACs and in primary NF2-negative meningioma lines. Moreover, we demonstrate that the dual mTORC1/mTORC2 inhibitor, AZD2014 is superior to rapamycin and PAK inhibitor FRAX597 in blocking proliferation of meningioma cells. Importantly, AZD2014 is currently in use in several clinical trials of cancer. Therefore, we believe that AZD2014 may provide therapeutic advantage over rapalogs for recurrent and progressive meningiomas.