Yang Yuanyuan, Zhang Xiaobai, Yi Li, Hou Zhenzhen, Chen Jiayu, Kou Xiaochen, Zhao Yanhong, Wang Hong, Sun Xiao-Fang, Jiang Cizhong, Wang Yixuan, Gao Shaorong
Clinical and Translational Research Center of Shanghai First Maternity & Infant Hospital, School of Life Sciences and Technology, Tongji University, Shanghai, People's Republic of China.
Key Laboratory for Major Obstetric Diseases of Guangdong Province, The Third Affiliated Hospital, Guangzhou Medical University, Guangdong, People's Republic of China.
Stem Cells Transl Med. 2016 Jan;5(1):8-19. doi: 10.5966/sctm.2015-0157. Epub 2015 Dec 16.
Conventional primed human embryonic stem cells and induced pluripotent stem cells (iPSCs) exhibit molecular and biological characteristics distinct from pluripotent stem cells in the naïve state. Although naïve pluripotent stem cells show much higher levels of self-renewal ability and multidifferentiation capacity, it is unknown whether naïve iPSCs can be generated directly from patient somatic cells and will be superior to primed iPSCs. In the present study, we used an established 5i/L/FA system to directly reprogram fibroblasts of a patient with β-thalassemia into transgene-free naïve iPSCs with molecular signatures of ground-state pluripotency. Furthermore, these naïve iPSCs can efficiently produce cross-species chimeras. Importantly, using the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 nuclease genome editing system, these naïve iPSCs exhibit significantly improved gene-correction efficiencies compared with the corresponding primed iPSCs. Furthermore, human naïve iPSCs could be directly generated from noninvasively collected urinary cells, which are easily acquired and thus represent an excellent cell resource for further clinical trials. Therefore, our findings demonstrate the feasibility and superiority of using patient-specific iPSCs in the naïve state for disease modeling, gene editing, and future clinical therapy.
In the present study, transgene-free naïve induced pluripotent stem cells (iPSCs) directly converted from the fibroblasts of a patient with β-thalassemia in a defined culture system were generated. These naïve iPSCs, which show ground-state pluripotency, exhibited significantly improved single-cell cloning ability, recovery capacity, and gene-targeting efficiency compared with conventional primed iPSCs. These results provide an improved strategy for personalized treatment of genetic diseases such as β-thalassemia.
传统的经预诱导的人类胚胎干细胞和诱导多能干细胞（iPSC）表现出与原始状态下的多能干细胞不同的分子和生物学特征。尽管原始多能干细胞表现出更高水平的自我更新能力和多向分化能力，但尚不清楚能否直接从患者体细胞产生原始iPSC，以及其是否优于经预诱导的iPSC。在本研究中，我们使用已建立的5i/L/FA系统，将一名β地中海贫血患者的成纤维细胞直接重编程为无转基因的具有原始多能性分子特征的原始iPSC。此外，这些原始iPSC能够高效产生跨物种嵌合体。重要的是，使用成簇规律间隔短回文重复序列（CRISPR）/CRISPR相关蛋白9核酸酶基因组编辑系统，与相应的经预诱导的iPSC相比，这些原始iPSC表现出显著提高的基因校正效率。此外，人类原始iPSC可直接从非侵入性收集的尿液细胞中产生，尿液细胞易于获取，因此是进一步临床试验的优质细胞资源。因此，我们的研究结果证明了使用原始状态的患者特异性iPSC进行疾病建模、基因编辑和未来临床治疗的可行性和优越性。
在本研究中，在特定培养系统中从一名β地中海贫血患者的成纤维细胞直接转化产生了无转基因的原始诱导多能干细胞（iPSC）。这些表现出原始多能性的原始iPSC，与传统的经预诱导的iPSC相比，表现出显著提高的单细胞克隆能力、恢复能力和基因靶向效率。这些结果为β地中海贫血等遗传性疾病的个性化治疗提供了一种改进策略。
