Single-cell signaling network profiling during redox stress reveals dynamic redox regulation in immune cells

Yi-Chuan Wang, Ping-Hsun Wu, Wen-Chieh Ting, Yi-Fu Wang, Ming-Han Yang, Tung-Hung Su, Jia-Ying Su, Hsun-I Sun, Wei-Min Huang, Pei-Ling Tsai, Gerlinde Wernig, Ping-Chih Ho, Limei Wang, Chen-Tu Wu, Yih-Leong Chang, Tseng-Cheng Chen, Tzu-Ching Meng, Yao-Ming Chang, Shih-Lei Lai, Chia-Wei Li, Tai-Ming Ko, Kai-Chien Yang, Ya-Jen Chang, Yijuang Chern, Mei-Chuan Kuo, Yen-Tsung Huang, Yi-Shiuan Tzeng, Jih-Luh Tang and Shih-Yu Chen ()
Additional contact information
Yi-Chuan Wang: National Yang Ming Chiao Tung University
Ping-Hsun Wu: Kaohsiung Medical University
Wen-Chieh Ting: National Taiwan University
Yi-Fu Wang: Academia Sinica
Ming-Han Yang: Academia Sinica
Tung-Hung Su: National Taiwan University Hospital
Jia-Ying Su: Academia Sinica
Hsun-I Sun: National Taiwan University
Wei-Min Huang: Retain Biotech
Pei-Ling Tsai: Retain Biotech
Gerlinde Wernig: Stanford University School of Medicine
Ping-Chih Ho: University of Lausanne
Limei Wang: University of Lausanne
Chen-Tu Wu: National Taiwan University Hospital and National Taiwan University College of Medicine
Yih-Leong Chang: National Taiwan University Hospital and National Taiwan University College of Medicine
Tseng-Cheng Chen: National Taiwan University Hospital and National Taiwan University College of Medicine
Tzu-Ching Meng: Academia Sinica
Yao-Ming Chang: Academia Sinica
Shih-Lei Lai: Academia Sinica
Chia-Wei Li: Academia Sinica
Tai-Ming Ko: Academia Sinica
Kai-Chien Yang: Academia Sinica
Ya-Jen Chang: Academia Sinica
Yijuang Chern: Academia Sinica
Mei-Chuan Kuo: Kaohsiung Medical University
Yen-Tsung Huang: Academia Sinica
Yi-Shiuan Tzeng: Academia Sinica
Jih-Luh Tang: National Taiwan University
Shih-Yu Chen: Academia Sinica

Nature Communications, 2025, vol. 16, issue 1, 1-18

Abstract: Abstract 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.

Date: 2025
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DOI: 10.1038/s41467-025-60727-z

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