A new class of pluripotent stem cell cytotoxic small molecules.
A new class of pluripotent stem cell cytotoxic small molecules.
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DOI：
10.1371/journal.pone.0085039
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Lee JM
中科院分区：
文献类型：
--
作者：
Richards M;Phoon CW;Goh GT;Seng EK;Guo XM;Tan CM;Chan WK;Lee JM
多能干细胞（PSC）衍生的细胞替代疗法中的主要问题是由污染的未分化细胞形成畸胎瘤的风险。从分化培养物中去除未分化细胞是PSC细胞疗法在临床环境中安全部署之前的重要步骤。我们报告了一组新的小分子，对PSC的细胞毒性。我们的数据表明，这些分子具有特异性和有效性，可以快速根除未分化细胞。利用混合PSC和原代人神经元和心肌细胞培养物的实验证明，可以获得高达6倍的特化细胞富集，而不会对细胞活力和功能产生不利影响。合成了几种结构变体以鉴定关键官能团并提高特异性和有效性。比较微阵列分析和随后的RNA敲除研究揭示了PERK/ATF 4/DDIT 3 ER应激途径的参与。令人惊讶的是，ER应激诱导后的细胞死亡与PSC中内源性ROS水平的伴随降低相关。在移植前用这些分子处理的未分化细胞不能在SCID小鼠中形成畸胎瘤。此外，这些分子对斑马鱼胚胎保持无毒和无致畸性，表明它们可以安全地在体内使用。
A major concern in Pluripotent Stem Cell (PSC)-derived cell replacement therapy is the risk of teratoma formation from contaminating undifferentiated cells. Removal of undifferentiated cells from differentiated cultures is an essential step before PSC-based cell therapies can be safely deployed in a clinical setting. We report a group of novel small molecules that are cytotoxic to PSCs. Our data indicates that these molecules are specific and potent in their activity allowing rapid eradication of undifferentiated cells. Experiments utilizing mixed PSC and primary human neuronal and cardiomyocyte cultures demonstrate that up to a 6-fold enrichment for specialized cells can be obtained without adversely affecting cell viability and function. Several structural variants were synthesized to identify key functional groups and to improve specificity and efficacy. Comparative microarray analysis and ensuing RNA knockdown studies revealed involvement of the PERK/ATF4/DDIT3 ER stress pathway. Surprisingly, cell death following ER stress induction was associated with a concomitant decrease in endogenous ROS levels in PSCs. Undifferentiated cells treated with these molecules preceding transplantation fail to form teratomas in SCID mice. Furthermore, these molecules remain non-toxic and non-teratogenic to zebrafish embryos suggesting that they may be safely used in vivo.
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影响因子：
3.7
作者：
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通讯作者：
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影响因子：
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通讯作者：
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