OVOL1 Influences the Determination and Expansion of iPSC Reprogramming Intermediates
OVOL1 Influences the Determination and Expansion of iPSC Reprogramming Intermediates
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DOI：
10.1016/j.stemcr.2018.12.008
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发表时间：
2019-02-12
影响因子：
5.9
通讯作者：
Woltjen, Knut
中科院分区：
文献类型：
--
作者：
Kagawa, Harunobu;Shimamoto, Ren;Woltjen, Knut
在体细胞重编程为诱导多能干细胞(IPSCs)的过程中，成纤维细胞经历了动态的分子变化，包括间充质向上皮的转变(MET)和获得多能性；这些过程受到山中因子化学计量比的影响。例如，在早期的重新编程中，高水平的KLF4与功能未定义的、瞬时表达的MET基因的诱导相关。在这里，我们确定了细胞表面蛋白TROP2作为高KLF4条件下瞬时MET诱导细胞的标志。我们观察到表达多能标记SSEA-1(+)的细胞主要来自TROP2(+)组分。在CRISPR/Cas9介导的MET基因候选筛选中，我们使用TROP2作为标记，我们鉴定了转录因子OVOL1是一种潜在的调节以非IPSC MET基因表达和低细胞增殖为特征的可选择的上皮细胞命运的因子。我们的研究揭示了重新编程因子化学计量比如何改变中间细胞命运的光谱，最终影响重新编程的结果。
During somatic cell reprogramming to induced pluripotent stem cells (iPSCs), fibroblasts undergo dynamic molecular changes, including a mesenchymal-to-epithelial transition (MET) and gain of pluripotency; processes that are influenced by Yamanaka factor stoichiometry. For example, in early reprogramming, high KLF4 levels are correlated with the induction of functionally undefined, transiently expressed MET genes. Here, we identified the cell-surface protein TROP2 as a marker for cells with transient MET induction in the high-KLF4 condition. We observed the emergence of cells expressing the pluripotency marker SSEA-1(+) mainly from within the TROP2(+) fraction. Using TROP2 as a marker in CRISPR/Cas9-mediated candidate screening of MET genes, we identified the transcription factor OVOL1 as a potential regulator of an alternative epithelial cell fate characterized by the expression of non-iPSC MET genes and low cell proliferation. Our study sheds light on how reprogramming factor stoichiometry alters the spectrum of intermediate cell fates, ultimately influencing reprogramming outcomes.