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
- Shih-Yu Chen
2025-07-01
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.