Liu Yang, Yang Yang, Suo Yangyang, Li Chuan, Chen Min, Zheng Shuwen, Li Hao, Tang Chengcheng, Fan Nana, Lan Ting, Zhou Jizeng, Li Yingying, Wang Jiaowei, Chen Huangyao, Zou Qingjian, Lai Liangxue
School of Life Sciences, University of Science and Technology of China, Hefei 230026, China.
Guangdong Provincial Key Laboratory of Large Animal Models for Biomedicine, School of Biotechnology and Health Sciences, Wuyi University, Jiangmen 529020, China.
Mol Ther Methods Clin Dev. 2022 Feb 1;24:332-341. doi: 10.1016/j.omtm.2022.01.014. eCollection 2022 Mar 10.
Pluripotent stem cells (PSCs) are promising in regenerative medicine. A major challenge of PSC therapy is the risk of teratoma formation because of the contamination of undifferentiated stem cells. Constitutive promoters or endogenous SOX2 promoters have been used to drive inducible caspase-9 () gene expression but cannot specifically eradicate undifferentiated PSCs. Here, we inserted gene into the endogenous OCT4 locus of human and mouse PSCs without affecting their pluripotency. A chemical inducer of dimerization (CID), AP1903, induced activation, which led to the apoptosis of specific undifferentiated PSCs and . Differentiated cell lineages survived because of the silence of the endogenous gene. Human and mouse PSCs were controllable when CID was administrated within 2 weeks after PSC injection in immunodeficient mice. However, an interval longer than 2 weeks caused teratoma formation and mouse death because a mass of somatic cells already differentiated from the PSCs. In conclusion, we have developed a specific and efficient PSC suicide system that will be of value in the clinical applications of PSC-based therapy.
多能干细胞（PSCs）在再生医学中前景广阔。PSC治疗的一个主要挑战是由于未分化干细胞的污染而形成畸胎瘤的风险。组成型启动子或内源性SOX2启动子已被用于驱动诱导型半胱天冬酶-9（）基因表达，但不能特异性根除未分化的PSCs。在这里，我们将基因插入到人和小鼠PSCs的内源性OCT4基因座中，而不影响它们的多能性。二聚化化学诱导剂（CID）AP1903诱导激活，导致特定未分化PSCs和的凋亡。由于内源性基因的沉默，分化的细胞谱系得以存活。当在免疫缺陷小鼠中注射PSC后2周内给予CID时，人和小鼠的PSCs是可控的。然而，超过2周的间隔会导致畸胎瘤形成和小鼠死亡，因为大量的体细胞已经从PSCs分化出来。总之，我们开发了一种特异性和高效的PSC自杀系统，这将在基于PSC的治疗的临床应用中具有价值。
