A molecular screening approach to identify and characterize inhibitors of glioblastoma stem cells.
A molecular screening approach to identify and characterize inhibitors of glioblastoma stem cells.
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DOI：
10.1158/1535-7163.mct-11-0268
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发表时间：
2011-10
影响因子：
5.7
通讯作者：
Kornblum HI
中科院分区：
文献类型：
--
作者：
Visnyei K;Onodera H;Damoiseaux R;Saigusa K;Petrosyan S;De Vries D;Ferrari D;Saxe J;Panosyan EH;Masterman-Smith M;Mottahedeh J;Bradley KA;Huang J;Sabatti C;Nakano I;Kornblum HI
多形性胶质母细胞瘤（GBM）是所有癌症中最致命的。GBM由肿瘤细胞的异质群体组成，其中肿瘤起始和治疗抗性亚群（在此称为GBM干细胞（GSC））已被鉴定为主要治疗靶标。在这里，我们描述了一种高通量的小分子筛选方法，使识别和表征的化合物，是有效的对GSC。该范例使用组织培养模型来富集来自人GBM切除的GSC，并将基于表型的筛选与基因靶特异性筛选相结合以用于化合物鉴定。我们使用了来自七个化学文库的31，624个小分子，我们根据它们对一组GSC富集培养物的影响以及它们对表达与临床结果呈负相关的基因模块表达的影响进行了表征和排名：MELK，ASPM，TOP 2A和FOXM 1b。在满足对所用细胞类型发挥差异效应的标准的11种化合物中，与非富集培养物相比，4种化合物通过抑制多种GSC富集培养物表现出选择性：依美汀、N-花生四烯酸多巴胺（NADA）、N-油酰多巴胺（OLDA）和N-棕榈酰多巴胺（PALDA）。ChemBridge化合物#5560509和#5256360抑制4种有丝分裂模块基因的表达。选择OLDA、依美汀和化合物#5560509和#5256360进行更详细的研究，并在体外自我更新测定和体内异种移植模型中抑制GSC。这些研究表明，我们的筛选策略提供了潜在的候选人，以及在更大规模的屏幕或屏幕涉及其他癌症类型的铅化合物识别的蓝图。
Glioblastoma multiforme (GBM) is amongst the most lethal of all cancers. GBM consist of a heterogeneous population of tumor cells amongst which a tumor initiating and treatment-resistant subpopulation, here termed GBM stem cells (GSC), have been identified as primary therapeutic targets. Here, we describe a high-throughput small molecule screening approach that enables the identification and characterization of chemical compounds that are effective against GSC. The paradigm uses a tissue culture model to enrich for GSC derived from human GBM resections and combines a phenotype-based screen with gene target-specific screens for compound identification. We used 31,624 small molecules from seven chemical libraries that we characterized and ranked based on their effect on a panel of GSC-enriched cultures as well as their effect on the expression of a module of genes whose expression negatively correlates with clinical outcome: MELK, ASPM, TOP2A and FOXM1b. Of the 11 compounds meeting criteria for exerting differential effects across cell types used, 4 compounds demonstrated selectivity by inhibiting multiple GSC-enriched cultures compared to non-enriched cultures: Emetine, N-Arachidonoyldopamine (NADA), N-Oleoyldopamine (OLDA), and N-Palmitoyldopamine (PALDA). ChemBridge compounds #5560509 and #5256360 inhibited the expression of the 4 mitotic module genes. OLDA, Emetine, and compounds #5560509 and #5256360 were chosen for more detailed study and inhibited GSC in self-renewal assays in vitro and in a xenograft model in vivo. These studies demonstrate that our screening strategy provides potential candidates as well as a blueprint for lead compound identification in larger scale screens or screens involving other cancer types.