A quantitative high-throughput screen for modulators of IL-6 signaling: a model for interrogating biological networks using chemical libraries.
A quantitative high-throughput screen for modulators of IL-6 signaling: a model for interrogating biological networks using chemical libraries.
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IL-6信号调节器的定量高通量屏幕：一种使用化学文库询问生物网络的模型。
DOI：
10.1039/b902021g
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发表时间：
2009-09
影响因子：
--
通讯作者：
Inglese J
中科院分区：
文献类型：
--
作者：
Johnson RL;Huang R;Jadhav A;Southall N;Wichterman J;MacArthur R;Xia M;Bi K;Printen J;Austin CP;Inglese J
小分子调节剂对于在分子水平上剖析和理解信号通路至关重要。白细胞介素6（IL-6）是一种细胞因子，通过JAK/STAT途径发出信号，并与癌症和炎症有关。为了鉴定该途径的调节剂，我们针对稳定表达与sis诱导元件融合的β-内酰胺酶报告基因的IL-6应答细胞系（SIE-bla细胞）筛选化学收集物。该测定针对1536孔微孔板格式进行了优化，并使用定量高通量筛选（qHTS）对11，693个小分子进行了筛选，qHTS是一种在多个浓度下测定化学文库以生成每种化合物的滴定响应曲线的方法。qHTS回收了564种具有良好拟合曲线的活性物质，这些活性物质聚集成32个不同的化学系列，包括13种活化剂和19种抑制剂。对qHTS数据的回顾性分析表明，1.5和7.7 μ M的单一浓度数据分别将35%和71%的qHTS活性物质评分为无活性，因此为假阴性。计数器筛选后，以确定荧光和非选择性系列，我们发现四个激活剂和一个抑制剂系列，调制SIE-bla细胞，但没有表现出类似的活性，在报告基因检测诱导EGF和缺氧。这些系列中的小分子将成为研究JAK/STAT激活介导的IL-6信号传导的有用工具化合物。
Small molecule modulators are critical for dissecting and understanding signaling pathways at the molecular level. Interleukin 6 (IL-6) is a cytokine that signals via the JAK/STAT pathway and is implicated in cancer and inflammation. To identify modulators of this pathway, we screened a chemical collection against an IL-6 responsive cell line stably expressing a beta-lactamase reporter gene fused to a sis-inducible element (SIE-bla cells). This assay was optimized for a 1536-well microplate format and screened against 11,693 small molecules using quantitative high-throughput screening (qHTS), a method that assays a chemical library at multiple concentrations to generate titration-response profiles for each compound. The qHTS recovered 564 actives with well-fit curves that clustered into 32 distinct chemical series of 13 activators and 19 inhibitors. A retrospective analysis of the qHTS data indicated that single concentration data at 1.5 and 7.7 uM scored 35 and 71% of qHTS actives, respectively, as inactive and were therefore false negatives. Following counter screens to identify fluorescent and nonselective series, we found four activator and one inhibitor series that modulated SIE-bla cells but did not show similar activity in reporter gene assays induced by EGF and hypoxia. Small molecules within these series will make useful tool compounds to investigate IL-6 signaling mediated by JAK/STAT activation.
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影响因子：
11.5
作者：
Duan, Zhenfeng;Bradner, James E.;Seiden, Michael V.
通讯作者：
Seiden, Michael V.
影响因子：
1.8
作者：
Davis, R. Eric;Zhang, Ya-Qin;Auld, Douglas S.
通讯作者：
Auld, Douglas S.
影响因子：
3.8
作者：
Duan, Z;Lamendola, DE;Seide, MV
通讯作者：
Seide, MV
影响因子：
5.4
作者：
ANDUS, T;GEIGER, T;HEINRICH, PC
通讯作者：
HEINRICH, PC
影响因子：
1.8
作者：
Chin, J;Adams, AD;Mitnaul, LJ
通讯作者：
Mitnaul, LJ