A novel quantitative high-throughput screen identifies drugs that both activate SUMO conjugation via the inhibition of microRNAs 182 and 183 and facilitate neuroprotection in a model of oxygen and glucose deprivation.
A novel quantitative high-throughput screen identifies drugs that both activate SUMO conjugation via the inhibition of microRNAs 182 and 183 and facilitate neuroprotection in a model of oxygen and glucose deprivation.
复制标题
DOI：
10.1177/0271678x15609939
复制
发表时间：
2016-02
期刊：
影响因子：
--
通讯作者：
Hallenbeck JM
中科院分区：
文献类型：
--
作者：
Bernstock JD;Lee YJ;Peruzzotti-Jametti L;Southall N;Johnson KR;Maric D;Volpe G;Kouznetsova J;Zheng W;Pluchino S;Hallenbeck JM
小泛素样修饰物（SUMO）的缀合/去缀合已显示与多种生理/病理条件相关。通过选择性控制整个SUMO化过程所提供的临床意义和表面上的治疗效用在缺血病理生理学中已经变得显而易见。在此，我们描述了一种新的定量高通量筛选（qHTS）系统的开发，该系统旨在鉴定能够通过调节/抑制microRNA（miRNA）-182家族成员来增加SUMO化的小分子。该测定采用用双荧光素酶-海肾荧光素酶报告系统稳定转染的SHSY 5 Y人神经母细胞瘤细胞系，用于鉴定miR-182或miR-183的特异性抑制剂。在这项研究中，我们已经确定了小分子能够诱导增加的全球结合的SUMO在SHSY 5 Y细胞和大鼠E18衍生的原代皮层神经元。通过体外缺血模型（氧/葡萄糖剥夺）证实了许多鉴定的化合物的保护作用。值得注意的是，这种测定可以很容易地重新利用，以允许高通量分析缺血病理生物学中其他相关miRNA的潜在可药用性。
The conjugation/de-conjugation of Small Ubiquitin-like Modifier (SUMO) has been shown to be associated with a diverse set of physiologic/pathologic conditions. The clinical significance and ostensible therapeutic utility offered via the selective control of the global SUMOylation process has become readily apparent in ischemic pathophysiology. Herein, we describe the development of a novel quantitative high-throughput screening (qHTS) system designed to identify small molecules capable of increasing SUMOylation via the regulation/inhibition of members of the microRNA (miRNA)-182 family. This assay employs a SHSY5Y human neuroblastoma cell line stably transfected with a dual firefly-Renilla luciferase reporter system for identification of specific inhibitors of either miR-182 or miR-183. In this study, we have identified small molecules capable of inducing increased global conjugation of SUMO in both SHSY5Y cells and rat E18-derived primary cortical neurons. The protective effects of a number of the identified compounds were confirmed via an in vitro ischemic model (oxygen/glucose deprivation). Of note, this assay can be easily repurposed to allow high-throughput analyses of the potential drugability of other relevant miRNA(s) in ischemic pathobiology.