Jiang Dacheng, Wang Yuchen, Chen Yanhao, Tian Cheng, Li Xin, Li Shuang, Gu Xiaosong, Jiang Chunping, Ding Qiurong
Shanghai Institute of Nutrition and Health, University of Chinese Academy of Sciences, Chinese Academy of Sciences, Shanghai, P. R. China.
Jinan Microecological Biomedicine Shandong Laboratory, Jinan, Shandong, P. R. China.
J Cell Physiol. 2025 Aug;240(8):e70083. doi: 10.1002/jcp.70083.
Mitochondria are crucial for cell fate determination, yet their roles in human pluripotent stem cell (hPSC) fate changes have remained underexplored. Here, we designed a CRISPR library targeting 661 mitochondrial proteins and identified the MPC (mitochondrial pyruvate carrier) as a critical regulator of hPSC self-renewal and pluripotency. Notably, MPC inhibition reduced hPSC self-renewal and endoderm differentiation while promoting mesoderm differentiation, with no effect on ectoderm differentiation, all mediated by influencing glycolytic acetyl-CoA production. Specifically, the decrease in acetyl-CoA following MPC inhibition affected histone acetylation in hPSCs, compromising self-renewal. In contrast, MPC inhibition did not impact histone acetylation in differentiated cells; instead, it reduced the acetylation of non-histone proteins-EP300 and SMAD2-thereby enhancing mesoderm differentiation and repressing endoderm differentiation, respectively. These findings suggest that distinct effector proteins respond to variations in acetyl-CoA levels at different developmental stages, leading to a context-dependent regulation of cell fate determination by glycolytic acetyl-CoA in hPSCs.
线粒体对于细胞命运的决定至关重要，然而它们在人类多能干细胞（hPSC）命运转变中的作用仍未得到充分探索。在此，我们设计了一个靶向661种线粒体蛋白的CRISPR文库，并确定线粒体丙酮酸载体（MPC）是hPSC自我更新和多能性的关键调节因子。值得注意的是，MPC抑制降低了hPSC的自我更新和内胚层分化，同时促进中胚层分化，对外胚层分化没有影响，所有这些都是通过影响糖酵解乙酰辅酶A的产生来介导的。具体而言，MPC抑制后乙酰辅酶A的减少影响了hPSC中的组蛋白乙酰化，损害了自我更新。相反，MPC抑制并不影响分化细胞中的组蛋白乙酰化；相反，它降低了非组蛋白蛋白EP300和SMAD2的乙酰化，从而分别增强了中胚层分化并抑制了内胚层分化。这些发现表明，不同的效应蛋白在不同发育阶段对乙酰辅酶A水平的变化做出反应，导致hPSC中糖酵解乙酰辅酶A对细胞命运决定的背景依赖性调节。
