A High-Throughput Screen Identifies 2,9-Diazaspiro[5.5]Undecanes as Inducers of the Endoplasmic Reticulum Stress Response with Cytotoxic Activity in 3D Glioma Cell Models.
A High-Throughput Screen Identifies 2,9-Diazaspiro[5.5]Undecanes as Inducers of the Endoplasmic Reticulum Stress Response with Cytotoxic Activity in 3D Glioma Cell Models.
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DOI：
10.1371/journal.pone.0161486
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Simeonov A
中科院分区：
文献类型：
--
作者：
Martinez NJ;Rai G;Yasgar A;Lea WA;Sun H;Wang Y;Luci DK;Yang SM;Nishihara K;Takeda S;Sagor M;Earnshaw I;Okada T;Mori K;Wilson K;Riggins GJ;Xia M;Grimaldi M;Jadhav A;Maloney DJ;Simeonov A
内质网(ER)参与钙信号和蛋白质折叠。内质网钙离子的耗竭和未折叠蛋白的积累激活了分子伴侣GRP78(葡萄糖调节蛋白78)，GRP78进而触发了内质网应激反应(ERSR)途径，旨在恢复内质网内稳态。然而，如果不能适应压力，就会导致细胞凋亡。我们和其他人已经表明，肿瘤细胞比正常细胞更容易受到ERSR诱导的凋亡的影响，这意味着ERSR是癌症治疗的潜在靶点。基于这些发现，我们开发了一种使用GRP78生物传感器来识别胶质瘤细胞中ERSR的小分子激活剂的方法。我们对大约425,000种化合物进行了定量高通量筛选(QHTS)，并制定了一套全面的正交二次分析面板，以进行严格的化合物验证。我们确定了新的ERSR激活剂，包括一种含有2，9-二氮螺[5.5]十一烷核心的化合物，它可以耗尽细胞内的钙存储并诱导几种癌细胞系的凋亡介导的细胞死亡，包括患者来源的和3D培养的胶质瘤细胞。这项研究表明，我们的筛选平台能够识别和分析具有细胞毒活性的ERSR诱导剂，并倡导在体内模型中表征这些化合物。
The endoplasmic reticulum (ER) is involved in Ca2+ signaling and protein folding. ER Ca2+ depletion and accumulation of unfolded proteins activate the molecular chaperone GRP78 (glucose-regulated protein 78) which in turn triggers the ER stress response (ERSR) pathway aimed to restore ER homeostasis. Failure to adapt to stress, however, results in apoptosis. We and others have shown that malignant cells are more susceptible to ERSR-induced apoptosis than their normal counterparts, implicating the ERSR as a potential target for cancer therapeutics. Predicated on these findings, we developed an assay that uses a GRP78 biosensor to identify small molecule activators of ERSR in glioma cells. We performed a quantitative high-throughput screen (qHTS) against a collection of ~425,000 compounds and a comprehensive panel of orthogonal secondary assays was formulated for stringent compound validation. We identified novel activators of ERSR, including a compound with a 2,9-diazaspiro[5.5]undecane core, which depletes intracellular Ca2+ stores and induces apoptosis-mediated cell death in several cancer cell lines, including patient-derived and 3D cultures of glioma cells. This study demonstrates that our screening platform enables the identification and profiling of ERSR inducers with cytotoxic activity and advocates for characterization of these compound in in vivo models.
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影响因子：
3.6
作者：
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通讯作者：
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