Central Laboratory, Guangxi Health Commission Key Laboratory of Glucose and Lipid Metabolism Disorders, The Second Affiliated Hospital of Guilin Medical University, Guilin 541000, China.
Clinical Medical Research Center, The Second Clinical Medical College, Jinan University, Shenzhen People's Hospital, Shenzhen 518020, China.
Biol Open. 2021 Feb 15;10(2):bio053348. doi: 10.1242/bio.053348.
Human induced pluripotent stem cells (iPSCs) are important source for regenerative medicine. However, the links between pluripotency and oncogenic transformation raise safety issues. To understand the characteristics of iPSC-derived cells at single-cell resolution, we directly reprogrammed two human iPSC lines into cardiomyocytes and collected cells from four time points during cardiac differentiation for single-cell sequencing. We captured 32,365 cells and identified five molecularly distinct clusters that aligned well with our reconstructed differentiation trajectory. We discovered a set of dynamic expression events related to the upregulation of oncogenes and the decreasing expression of tumor suppressor genes during cardiac differentiation, which were similar to the gain-of-function and loss-of-function patterns during oncogenesis. In practice, we characterized the dynamic expression of the and Yamanaka factor genes (, , and ), which were widely used for human iPSCs lines generation; and revealed the co-occurrence of overexpression and silencing in some of human iPSC-derived + cardiomyocytes. In summary, our oncogenic expression atlas is valuable for human iPSCs application and the single-cell resolution highlights the clues potentially associated with the carcinogenic risk of human iPSC-derived cells.
人类诱导多能干细胞（iPSCs）是再生医学的重要来源。然而，多能性与致癌转化之间的联系引发了安全问题。为了在单细胞分辨率水平上了解 iPSC 衍生细胞的特征，我们直接将两条人类 iPSC 系重编程为心肌细胞，并在心脏分化的四个时间点收集细胞进行单细胞测序。我们捕获了 32365 个细胞，并鉴定出五个分子上明显不同的簇，与我们重建的分化轨迹很好地吻合。我们发现了一组与心脏分化过程中致癌基因上调和肿瘤抑制基因表达下调相关的动态表达事件，这些事件与致癌过程中的功能获得和功能丧失模式相似。实际上，我们对广泛用于人类 iPSC 系生成的 和 Yamanaka 因子基因（、、和 ）的动态表达进行了表征；并揭示了一些人类 iPSC 衍生的 +心肌细胞中 过表达和 沉默的共发生。总之，我们的致癌表达图谱对于人类 iPSC 的应用具有重要价值，单细胞分辨率突出了与人类 iPSC 衍生细胞致癌风险相关的潜在线索。
