Expression data from human pluripotent stem cells and differentiated cells treated with pluripotent-specific inhibitors

The use of human pluripotent stem cells (hPSCs) in cell therapy is hindered by the tumorigenic risk from residual undifferentiated cells. Here we performed a high-throughput screen of over 52,000 small molecules, and identified 15 highly selective cytotoxic inhibitors of hPSCs (PluriSIns). Cellular and molecular analyses revealed that the most selective compound, PluriSIn #1, is a pluripotent-specific inhibitor of stearoyl-coA desaturase (SCD1), the key enzyme in the biosynthesis of monounsaturated fatty acids (MUFA). SCD1 inhibition in hPSCs induced ER stress, protein synthesis attenuation, and apoptosis of these cells, revealing that MUFA biosynthesis is crucial for their survival. PluriSIn #1 was also cytotoxic toward the ICM cells of mouse embryos, indicating that the dependence on SCD1 is inherent to the pluripotent state. Finally, application of PluriSIn #1 prevented teratoma formation from tumorigenic undifferentiated cells. Our novel method to eliminate undifferentiated cells from culture should thus increase the safety of hPSC-based treatments. Expression data from undifferentiated and differentiated human embryonic stem cells. Total RNA was isolated from undifferentiated human pluripotent stem cells grown on matrigel with mTeSR1 medium, or from early endodermal progenitor cells differentiated from human embryonic stem cells.

人类多能干细胞（human pluripotent stem cells, hPSCs）应用于细胞治疗时，常因残留未分化细胞存在致瘤风险，这一缺陷制约了其临床转化。本研究针对52000余种小分子化合物开展高通量筛选，成功鉴定出15种对hPSCs具有高度选择性的细胞毒性抑制剂（PluriSIns）。细胞及分子水平分析表明，选择性最优的化合物PluriSIn #1是一种多能状态特异性的硬脂酰辅酶A去饱和酶（stearoyl-coA desaturase, SCD1）抑制剂，而SCD1是单不饱和脂肪酸（monounsaturated fatty acids, MUFA）生物合成的关键酶。在hPSCs中抑制SCD1可诱发内质网应激（ER stress）、抑制蛋白质合成并诱导细胞凋亡，证实MUFA生物合成对hPSCs的存活具有不可或缺的作用。PluriSIn #1对小鼠胚胎内细胞团（ICM）细胞同样具有细胞毒性，提示对SCD1的依赖性是多能细胞状态的固有特征。进一步实验显示，使用PluriSIn #1可有效阻断致瘤性未分化细胞形成畸胎瘤。本研究建立的从培养体系中清除未分化细胞的新方法，将有助于提升基于hPSCs的治疗方案的安全性。本数据集包含未分化及分化人类胚胎干细胞的表达谱数据：总RNA分别提取自以mTeSR1培养基在基质胶上培养的未分化人类多能干细胞，以及由人类胚胎干细胞分化得到的早期内胚层祖细胞。