Identification of Oct4-activating compounds that enhance reprogramming efficiency
Identification of Oct4-activating compounds that enhance reprogramming efficiency
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DOI：
10.1073/pnas.1219181110
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发表时间：
2012-12-18
影响因子：
11.1
通讯作者：
Shi, Yanhong
中科院分区：
文献类型：
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作者：
Li, Wendong;Tian, E.;Shi, Yanhong
诱导多能干细胞（iPSC）的实际应用的障碍之一是低效率和缓慢的重编程过程。八聚体结合转录因子4（Oct 4）已被证明是胚胎干细胞（ESC）多能性的重要调节因子，也是重编程过程的关键。为了鉴定增强重编程效率的小分子，我们进行了基于细胞的化学文库的高通量筛选。其中一种被称为Oct 4激活化合物1（OAC 1）的化合物被发现可以激活Oct 4和Nanog启动子驱动的荧光素酶报告基因。此外，当与四重编程因子（Oct 4、Sox 2、c-Myc和Klf 4）一起沿着添加到重编程混合物中时，OAC 1增强iPSC重编程效率并加速重编程过程。OAC 1的两种结构类似物也激活了Oct 4和Nanog启动子，并增强了iPSC的形成。使用Oct 4活化化合物沿着与四重因子衍生的iPSC集落表现出典型的ESC形态、基因表达模式和发育潜力。OAC 1似乎以一种独特的方式增强重编程效率，不依赖于p53-p21通路的抑制或Wnt-β-连环蛋白信号传导的激活。OAC 1增加Oct 4-Nanog-Sox 2三联体和Tet 1的转录，Tet 1是已知参与DNA去甲基化的基因。
One of the hurdles for practical application of induced pluripotent stem cells (iPSC) is the low efficiency and slow process of reprogramming. Octamer-binding transcription factor 4 (Oct4) has been shown to be an essential regulator of embryonic stem cell (ESC) pluripotency and key to the reprogramming process. To identify small molecules that enhance reprogramming efficiency, we performed a cell-based high-throughput screening of chemical libraries. One of the compounds, termed Oct4-activating compound 1 (OAC1), was found to activate both Oct4 and Nanog promoter-driven luciferase reporter genes. Furthermore, when added to the reprogramming mixture along with the quartet reprogramming factors (Oct4, Sox2, c-Myc, and Klf4), OAC1 enhanced the iPSC reprogramming efficiency and accelerated the reprogramming process. Two structural analogs of OAC1 also activated Oct4 and Nanog promoters and enhanced iPSC formation. The iPSC colonies derived using the Oct4-activating compounds along with the quartet factors exhibited typical ESC morphology, gene-expression pattern, and developmental potential. OAC1 seems to enhance reprogramming efficiency in a unique manner, independent of either inhibition of the p53-p21 pathway or activation of the Wnt-beta-catenin signaling. OAC1 increases transcription of the Oct4-Nanog-Sox2 triad and Tet1, a gene known to be involved in DNA demethylation.