Identification of known drugs targeting the endoplasmic reticulum stress response
Identification of known drugs targeting the endoplasmic reticulum stress response
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DOI：
10.1007/s00216-015-8694-2
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发表时间：
2015-07-01
影响因子：
4.3
通讯作者：
Xia, Menghang
中科院分区：
文献类型：
--
作者：
Bi, Kun;Nishihara, Kana;Xia, Menghang
内质网（ER）是一种多功能细胞器，在细胞信号传导、发育和应激反应中起着核心作用。内质网稳态失调与人类疾病有关，如癌症、炎症和糖尿病。包括缺氧在内的多种应激刺激以及多种药物均可导致内质网应激反应。在本研究中，我们开发了一种稳定的内质网应激报告细胞系，该细胞系在葡萄糖调节蛋白78 kDa （GRP78）基因启动子中的内质网应激反应元件（ESRE）的控制下稳定表达β -内酰胺酶报告基因。该分析已被优化并小型化为1536孔板格式。为了确定临床使用的诱导内质网应激反应的药物，我们使用定量高通量筛选（qHTS）平台从NIH化学基因组学中心药物收集（NPC库）中筛选了大约2800种药物。从这项研究中，我们已经确定了几种已知的内质网应激诱导剂，如17-AAG（通过HSP90抑制），以及几种新的内质网应激诱导剂，如AMI-193和spiperone。进一步研究确定的药物对真核起始因子2 α (eIF2 α)磷酸化、x- box结合蛋白（XBP1）剪接和GRP78基因表达的影响。这些结果表明，使用qHTS方法从NPC文库中鉴定出的内质网应激诱导剂可以为这些药物的潜在治疗靶点提供新的线索。
The endoplasmic reticulum (ER), a multifunctional organelle, plays a central role in cellular signaling, development, and stress response. Dysregulation of ER homeostasis has been associated with human diseases, such as cancer, inflammation, and diabetes. A broad spectrum of stressful stimuli including hypoxia as well as a variety of pharmacological agents can lead to the ER stress response. In this study, we have developed a stable ER stress reporter cell line that stably expresses a beta-lactamase reporter gene under the control of the ER stress response element (ESRE) present in the glucose-regulated protein, 78 kDa (GRP78) gene promoter. This assay has been optimized and miniaturized into a 1536-well plate format. In order to identify clinically used drugs that induce ER stress response, we screened approximately 2800 drugs from the NIH Chemical Genomics Center Pharmaceutical Collection (NPC library) using a quantitative high-throughput screening (qHTS) platform. From this study, we have identified several known ER stress inducers, such as 17-AAG (via HSP90 inhibition), as well as several novel ER stress inducers such as AMI-193 and spiperone. The confirmed drugs were further studied for their effects on the phosphorylation of eukaryotic initiation factor 2 alpha (eIF2 alpha), the X-box-binding protein (XBP1) splicing, and GRP78 gene expression. These results suggest that the ER stress inducers identified from the NPC library using the qHTS approach could shed new lights on the potential therapeutic targets of these drugs.