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

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...

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Main Authors: 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
Format: Article
Language:English
Published: Nature Portfolio 2025-07-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-60727-z
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author 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
author_facet 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
author_sort Yi-Chuan Wang
collection DOAJ
description 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.
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publisher Nature Portfolio
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series Nature Communications
spelling doaj-art-d3606999b8d1404fb58ca23cabc5bb7e2025-08-20T03:45:33ZengNature PortfolioNature Communications2041-17232025-07-0116111810.1038/s41467-025-60727-zSingle-cell signaling network profiling during redox stress reveals dynamic redox regulation in immune cellsYi-Chuan Wang0Ping-Hsun Wu1Wen-Chieh Ting2Yi-Fu Wang3Ming-Han Yang4Tung-Hung Su5Jia-Ying Su6Hsun-I Sun7Wei-Min Huang8Pei-Ling Tsai9Gerlinde Wernig10Ping-Chih Ho11Limei Wang12Chen-Tu Wu13Yih-Leong Chang14Tseng-Cheng Chen15Tzu-Ching Meng16Yao-Ming Chang17Shih-Lei Lai18Chia-Wei Li19Tai-Ming Ko20Kai-Chien Yang21Ya-Jen Chang22Yijuang Chern23Mei-Chuan Kuo24Yen-Tsung Huang25Yi-Shiuan Tzeng26Jih-Luh Tang27Shih-Yu Chen28Program in Molecular Medicine, National Yang Ming Chiao Tung UniversityDivision of Nephrology, Department of Internal Medicine, Kaohsiung Medical University Hospital, Kaohsiung Medical UniversityTai-Chen Stem Cell Therapy Center, National Taiwan UniversityInstitute of Biomedical Sciences, Academia SinicaInstitute of Biomedical Sciences, Academia SinicaDivision of Gastroenterology and Hepatology, Department of Internal Medicine, National Taiwan University HospitalInstitute of Statistical Science, Academia SinicaTai-Chen Stem Cell Therapy Center, National Taiwan UniversityRetain BiotechRetain BiotechDepartment of Pathology, Institute of Stem Cell Biology and Regenerative Medicine, Stanford University School of MedicineDepartment of Oncology, University of LausanneDepartment of Oncology, University of LausanneDepartment of Pathology, National Taiwan University Cancer Center, National Taiwan University Hospital and National Taiwan University College of MedicineDepartment of Pathology, National Taiwan University Cancer Center, National Taiwan University Hospital and National Taiwan University College of MedicineDepartment of Otolaryngology, National Taiwan University Hospital and National Taiwan University College of MedicineInstitute of Biological Chemistry, Academia SinicaInstitute of Biomedical Sciences, Academia SinicaInstitute of Biomedical Sciences, Academia SinicaInstitute of Biomedical Sciences, Academia SinicaInstitute of Biomedical Sciences, Academia SinicaInstitute of Biomedical Sciences, Academia SinicaInstitute of Biomedical Sciences, Academia SinicaInstitute of Biomedical Sciences, Academia SinicaDivision of Nephrology, Department of Internal Medicine, Kaohsiung Medical University Hospital, Kaohsiung Medical UniversityInstitute of Statistical Science, Academia SinicaInstitute of Biomedical Sciences, Academia SinicaTai-Chen Stem Cell Therapy Center, National Taiwan UniversityInstitute of Biomedical Sciences, Academia SinicaAbstract 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.https://doi.org/10.1038/s41467-025-60727-z
spellingShingle 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
Single-cell signaling network profiling during redox stress reveals dynamic redox regulation in immune cells
Nature Communications
title Single-cell signaling network profiling during redox stress reveals dynamic redox regulation in immune cells
title_full Single-cell signaling network profiling during redox stress reveals dynamic redox regulation in immune cells
title_fullStr Single-cell signaling network profiling during redox stress reveals dynamic redox regulation in immune cells
title_full_unstemmed Single-cell signaling network profiling during redox stress reveals dynamic redox regulation in immune cells
title_short Single-cell signaling network profiling during redox stress reveals dynamic redox regulation in immune cells
title_sort single cell signaling network profiling during redox stress reveals dynamic redox regulation in immune cells
url https://doi.org/10.1038/s41467-025-60727-z
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