Identification of Specific Pluripotent Stem Cell Death-Inducing Small Molecules by Chemical Screening
Identification of Specific Pluripotent Stem Cell Death-Inducing Small Molecules by Chemical Screening
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DOI：
10.1007/s12015-011-9248-4
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发表时间：
2012-03-01
影响因子：
4.8
通讯作者：
Alberto Carrodeguas, Jose
中科院分区：
文献类型：
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作者：
Conesa, Celia;Doss, Michael Xavier;Alberto Carrodeguas, Jose
胚胎和诱导多能干细胞治疗退行性疾病的潜在应用涉及纯体细胞，不含致瘤性未分化胚胎和诱导多能干细胞。在具有药理潜力的复杂化学物质集合中，我们期望找到能够诱导特异性多能干细胞死亡的分子，这些分子可用于某些细胞治疗设置，以消除未分化的细胞。因此，我们筛选了一个包含1120种小化学物质的化学文库，以确定在未分化的小鼠胚胎干细胞（ESCs）中特异性诱导凋亡细胞死亡的化合物。有趣的是，目前作为临床批准药物使用的三种化合物，即去甲替林、氯化苄索铵和甲基氯化苄索铵，对ESCs和小鼠胚胎成纤维细胞（mef）的细胞活力产生了不同的影响。去甲替林在mef中诱导细胞凋亡，而在ESCs中不诱导，而苄索溴铵和甲基苄索氯铵则表现出相反的效果。去甲替林是一种三环抗抑郁药，也被认为是线粒体通透性转变的有效抑制剂，线粒体通透性转变是凋亡过程中线粒体膜通透化的两种主要机制之一。苄索氯铵和甲基苄索氯铵是一种广谱抗菌季铵盐，也被描述为抗癌剂。与原代人皮肤成纤维细胞和已建立的人成纤维细胞系相比，苯并氯铵在人诱导多能干细胞（hiPSCs）中也观察到类似的作用。人类成纤维细胞和hiPSCs对去甲替林具有相似的耐药性，尽管表现不同。我们的研究结果表明，ESCs、hiPSCs和成纤维细胞对某些化合物的敏感性存在差异，这可能在干细胞治疗中具有重要的应用价值，通过从干细胞来源的体细胞中去除未分化的多能干细胞，以防止移植后肿瘤的形成，用于治疗退行性疾病。
A potential application of embryonic and inducible pluripotent stem cells for the therapy of degenerative diseases involves pure somatic cells, free of tumorigenic undifferentiated embryonic and inducible pluripotent stem cells. In complex collections of chemicals with pharmacological potential we expect to find molecules able to induce specific pluripotent stem cell death, which could be used in some cell therapy settings to eliminate undifferentiated cells. Therefore, we have screened a chemical library of 1120 small chemicals to identify compounds that induce specifically apoptotic cell death in undifferentiated mouse embryonic stem cells (ESCs). Interestingly, three compounds currently used as clinically approved drugs, nortriptyline, benzethonium chloride and methylbenzethonium chloride, induced differential effects in cell viability in ESCs versus mouse embryonic fibroblasts (MEFs). Nortriptyline induced apoptotic cell death in MEFs but not in ESCs, whereas benzethonium and methylbenzethonium chloride showed the opposite effect. Nortriptyline, a tricyclic antidepressant, has also been described as a potent inhibitor of mitochondrial permeability transition, one of two major mechanisms involved in mitochondrial membrane permeabilization during apoptosis. Benzethonium chloride and methylbenzethonium chloride are quaternary ammonium salts used as antimicrobial agents with broad spectrum and have also been described as anticancer agents. A similar effect of benzethonium chloride was observed in human induced pluripotent stem cells (hiPSCs) when compared to both primary human skin fibroblasts and an established human fibroblast cell line. Human fibroblasts and hiPSCs were similarly resistant to nortriptyline, although with a different behavior. Our results indicate differential sensitivity of ESCs, hiPSCs and fibroblasts to certain chemical compounds, which might have important applications in the stem cell-based therapy by eliminating undifferentiated pluripotent stem cells from stem cell-derived somatic cells to prevent tumor formation after transplantation for therapy of degenerative diseases.