Wang Yi-Chuan, Wu Ping-Hsun, Ting Wen-Chieh, Wang Yi-Fu, Yang Ming-Han, Su Tung-Hung, Su Jia-Ying, Sun Hsun-I, Huang Wei-Min, Tsai Pei-Ling, Wernig Gerlinde, Ho Ping-Chih, Wang Limei, Wu Chen-Tu, Chang Yih-Leong, Chen Tseng-Cheng, Meng Tzu-Ching, Chang Yao-Ming, Lai Shih-Lei, Li Chia-Wei, Ko Tai-Ming, Yang Kai-Chien, Chang Ya-Jen, Chern Yijuang, Kuo Mei-Chuan, Huang Yen-Tsung, Tzeng Yi-Shiuan, Tang Jih-Luh, Chen Shih-Yu
Program in Molecular Medicine, National Yang Ming Chiao Tung University, Taipei, Taiwan.
Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan.
Nat Commun. 2025 Jul 1;16(1):5600. doi: 10.1038/s41467-025-60727-z.
In eukaryotic cells, reactive oxygen species (ROS) serve as crucial signaling components. ROS are potentially toxic, so constant adjustments are needed to maintain cellular health. Here we describe a single-cell, mass cytometry-based method that we call signaling network under redox stress profiling (SN-ROP) to monitor dynamic changes in redox-related pathways during redox stress. SN-ROP quantifies ROS transporters, enzymes, oxidative stress products and associated signaling pathways to provide information on cellular redox regulation. Applied to diverse cell types and conditions, SN-ROP reveals unique redox patterns and dynamics including coordinated shifts in CD8 T cells upon antigen stimulation as well as variations in CAR-T cell persistence. Furthermore, SN-ROP analysis uncovers environmental factors such as hypoxia and T cell exhaustion for influencing redox balance, and also reveals distinct features in patients on hemodialysis. Our findings thus support the use of SN-ROP to elucidate intricate redox networks and their implications in immune cell function and disease.
在真核细胞中，活性氧（ROS）是关键的信号传导成分。ROS具有潜在毒性，因此需要不断调节以维持细胞健康。在此，我们描述了一种基于单细胞质谱流式细胞术的方法，我们称之为氧化还原应激分析下的信号网络（SN-ROP），用于监测氧化还原应激期间氧化还原相关途径的动态变化。SN-ROP对ROS转运蛋白、酶、氧化应激产物及相关信号通路进行定量，以提供细胞氧化还原调节的信息。应用于多种细胞类型和条件下，SN-ROP揭示了独特的氧化还原模式和动态变化，包括抗原刺激后CD8 T细胞的协同变化以及CAR-T细胞持久性的差异。此外，SN-ROP分析揭示了诸如缺氧和T细胞耗竭等影响氧化还原平衡的环境因素，还揭示了血液透析患者的独特特征。因此，我们的研究结果支持使用SN-ROP来阐明复杂的氧化还原网络及其在免疫细胞功能和疾病中的意义。
