Lv Jing, Yi Ying, Qi Yan, Yan Chenchao, Jin Wenwen, Meng Liming, Zhang Donghui, Jiang Wei
Department of Biological Repositories, Frontier Science Center for Immunology and Metabolism, Medical Research Institute, Zhongnan Hospital of Wuhan University, Wuhan University, Wuhan, 430071, China.
State Key Laboratory of Biocatalysis and Enzyme Engineering, School of Life Science, Hubei University, Wuhan, 430062, China.
Cell Death Discov. 2022 Feb 17;8(1):69. doi: 10.1038/s41420-022-00867-z.
Cellular organelles play fundamental roles in almost all cell behaviors. Mitochondria have been reported to be functionally linked to various biological processes, including reprogramming and pluripotency maintenance. However, very little about the role of mitochondria has been revealed in human early development and lineage specification. Here, we reported the characteristics and function of mitochondria during human definitive endoderm differentiation. Using a well-established differentiation system, we first investigated the change of mitochondrial morphology by comparing undifferentiated pluripotent stem cells, the intermediate mesendoderm cells, and differentiated endoderm cells, and found that mitochondria were gradually elongated and matured along differentiation. We further analyzed the expression pattern of mitochondria-related genes by RNA-seq, indicating that mitochondria became active during differentiation. Supporting this notion, the production of adenosine triphosphate (ATP) and reactive oxygen species (ROS) was increased as well. Functionally, we utilized chemicals and genome editing techniques, which could interfere with mitochondrial homeostasis, to determine the role of mitochondria in human endoderm differentiation. Treatment with mitochondrial inhibitors, or genetic depletion of mitochondrial transcription factor A (TFAM), significantly reduced the differentiation efficiency of definitive endoderm. In addition, the defect in endoderm differentiation due to dysfunctional mitochondria could be restored to some extent by the addition of ATP. Moreover, the clearance of excessive ROS due to dysfunctional mitochondria by N-acetylcysteine (NAC) improved the differentiation as well. We further found that ATP and NAC could partially replace the growth factor activin A for definitive endoderm differentiation. Our study illustrates the essential role of mitochondria during human endoderm differentiation through providing ATP and regulating ROS levels, which may provide new insight for metabolic regulation of cell fate determination.
细胞器在几乎所有细胞行为中都发挥着重要作用。据报道，线粒体在功能上与多种生物学过程相关联，包括重编程和多能性维持。然而，线粒体在人类早期发育和谱系特化中的作用却鲜为人知。在此，我们报道了人类定形内胚层分化过程中线粒体的特征和功能。利用成熟的分化系统，我们首先通过比较未分化的多能干细胞、中间中胚层细胞和分化的内胚层细胞，研究了线粒体形态的变化，发现线粒体在分化过程中逐渐伸长并成熟。我们进一步通过RNA测序分析了线粒体相关基因的表达模式，表明线粒体在分化过程中变得活跃。支持这一观点的是，三磷酸腺苷（ATP）和活性氧（ROS）的产生也增加了。在功能上，我们利用能够干扰线粒体稳态的化学物质和基因组编辑技术，来确定线粒体在人类内胚层分化中的作用。用线粒体抑制剂处理或线粒体转录因子A（TFAM）的基因缺失，显著降低了定形内胚层的分化效率。此外，通过添加ATP，线粒体功能障碍导致的内胚层分化缺陷在一定程度上可以得到恢复。而且，用N-乙酰半胱氨酸（NAC）清除线粒体功能障碍产生的过量ROS也改善了分化。我们还发现，ATP和NAC可以部分替代生长因子激活素A促进定形内胚层的分化。我们的研究通过提供ATP和调节ROS水平，阐明了线粒体在人类内胚层分化中的重要作用，这可能为细胞命运决定的代谢调控提供新的见解。
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