N-deglycosylation targeting chimera (DGlyTAC): a strategy for immune checkpoint proteins inactivation by specifically removing N-glycan

Abstract Among the leading methods for triggering therapeutic anti-cancer immunity is the inhibition of immune checkpoint pathways. N-glycosylation is found to be essential for the function of various immune checkpoint proteins, playing a critical role in their stability and interaction with immune...

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Main Authors: Li Li, Jiajia Wu, Weiqian Cao, Wei Zhang, Qi Wu, Yaxu Li, Yanrong Yang, Zezhi Shan, Zening Zheng, Xin Ge, Liang Lin, Ping Wang
Format: Article
Language:English
Published: Nature Publishing Group 2025-04-01
Series:Signal Transduction and Targeted Therapy
Online Access:https://doi.org/10.1038/s41392-025-02219-6
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author Li Li
Jiajia Wu
Weiqian Cao
Wei Zhang
Qi Wu
Yaxu Li
Yanrong Yang
Zezhi Shan
Zening Zheng
Xin Ge
Liang Lin
Ping Wang
author_facet Li Li
Jiajia Wu
Weiqian Cao
Wei Zhang
Qi Wu
Yaxu Li
Yanrong Yang
Zezhi Shan
Zening Zheng
Xin Ge
Liang Lin
Ping Wang
author_sort Li Li
collection DOAJ
description Abstract Among the leading methods for triggering therapeutic anti-cancer immunity is the inhibition of immune checkpoint pathways. N-glycosylation is found to be essential for the function of various immune checkpoint proteins, playing a critical role in their stability and interaction with immune cells. Removing the N-glycans of these proteins seems to be an alternative therapy, but there is a lack of a de-N-glycosylation technique for target protein specificity, which limits its clinical application. Here, we developed a novel technique for specifically removing N-glycans from a target protein on the cell surface, named deglycosylation targeting chimera (DGlyTAC), which employs a fusing protein consisting of Peptide-N-glycosidase F (PNGF) and target-specific nanobody/affibody (Nb/Af). The DGlyTAC technique was developed to target a range of glycosylated surface proteins, especially these immune checkpoints—CD24, CD47, and PD-L1, which minimally affected the overall N-glycosylation landscape and the N-glycosylation of other representative membrane proteins, ensuring high specificity and minimal off-target effects. Importantly, DGlyTAC technique was successfully applied to lead inactivation of these immune checkpoints, especially PD-L1, and showed more potential in cancer immunotherapy than inhibitors. Finally, PD-L1 targeted DGlyTAC showed therapeutic effects on several tumors in vivo, even better than PD-L1 antibody. Overall, we created a novel target-specific N-glysocylation erasing technique that establishes a modular strategy for directing membrane proteins inactivation, with broad implications on tumor immune therapeutics.
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publishDate 2025-04-01
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spelling doaj-art-b4e7a6658eff4e2b8f8b5028fba4a3df2025-08-20T03:52:24ZengNature Publishing GroupSignal Transduction and Targeted Therapy2059-36352025-04-0110111510.1038/s41392-025-02219-6N-deglycosylation targeting chimera (DGlyTAC): a strategy for immune checkpoint proteins inactivation by specifically removing N-glycanLi Li0Jiajia Wu1Weiqian Cao2Wei Zhang3Qi Wu4Yaxu Li5Yanrong Yang6Zezhi Shan7Zening Zheng8Xin Ge9Liang Lin10Ping Wang11Tongji University Cancer Center, Shanghai Tenth People’s Hospital, School of Medicine, Tongji UniversityState Key Laboratory of Chemical Biology, Shanghai Institute of Organic Chemistry, Chinese Academy of SciencesInstitutes of Biomedical Sciences, Fudan UniversityInstitutes of Biomedical Sciences, Fudan UniversityTongji University Cancer Center, Shanghai Tenth People’s Hospital, School of Medicine, Tongji UniversityShanghai Frontiers Science Center of Nanocatalytic Medicine, School of Medicine, Tongji UniversityShanghai Frontiers Science Center of Nanocatalytic Medicine, School of Medicine, Tongji UniversityDepartment of Colorectal Surgery, Fudan University Shanghai Cancer CenterShanghai Frontiers Science Center of Nanocatalytic Medicine, School of Medicine, Tongji UniversityTongji University Cancer Center, Shanghai Tenth People’s Hospital, School of Medicine, Tongji UniversityState Key Laboratory of Chemical Biology, Shanghai Institute of Organic Chemistry, Chinese Academy of SciencesTongji University Cancer Center, Shanghai Tenth People’s Hospital, School of Medicine, Tongji UniversityAbstract Among the leading methods for triggering therapeutic anti-cancer immunity is the inhibition of immune checkpoint pathways. N-glycosylation is found to be essential for the function of various immune checkpoint proteins, playing a critical role in their stability and interaction with immune cells. Removing the N-glycans of these proteins seems to be an alternative therapy, but there is a lack of a de-N-glycosylation technique for target protein specificity, which limits its clinical application. Here, we developed a novel technique for specifically removing N-glycans from a target protein on the cell surface, named deglycosylation targeting chimera (DGlyTAC), which employs a fusing protein consisting of Peptide-N-glycosidase F (PNGF) and target-specific nanobody/affibody (Nb/Af). The DGlyTAC technique was developed to target a range of glycosylated surface proteins, especially these immune checkpoints—CD24, CD47, and PD-L1, which minimally affected the overall N-glycosylation landscape and the N-glycosylation of other representative membrane proteins, ensuring high specificity and minimal off-target effects. Importantly, DGlyTAC technique was successfully applied to lead inactivation of these immune checkpoints, especially PD-L1, and showed more potential in cancer immunotherapy than inhibitors. Finally, PD-L1 targeted DGlyTAC showed therapeutic effects on several tumors in vivo, even better than PD-L1 antibody. Overall, we created a novel target-specific N-glysocylation erasing technique that establishes a modular strategy for directing membrane proteins inactivation, with broad implications on tumor immune therapeutics.https://doi.org/10.1038/s41392-025-02219-6
spellingShingle Li Li
Jiajia Wu
Weiqian Cao
Wei Zhang
Qi Wu
Yaxu Li
Yanrong Yang
Zezhi Shan
Zening Zheng
Xin Ge
Liang Lin
Ping Wang
N-deglycosylation targeting chimera (DGlyTAC): a strategy for immune checkpoint proteins inactivation by specifically removing N-glycan
Signal Transduction and Targeted Therapy
title N-deglycosylation targeting chimera (DGlyTAC): a strategy for immune checkpoint proteins inactivation by specifically removing N-glycan
title_full N-deglycosylation targeting chimera (DGlyTAC): a strategy for immune checkpoint proteins inactivation by specifically removing N-glycan
title_fullStr N-deglycosylation targeting chimera (DGlyTAC): a strategy for immune checkpoint proteins inactivation by specifically removing N-glycan
title_full_unstemmed N-deglycosylation targeting chimera (DGlyTAC): a strategy for immune checkpoint proteins inactivation by specifically removing N-glycan
title_short N-deglycosylation targeting chimera (DGlyTAC): a strategy for immune checkpoint proteins inactivation by specifically removing N-glycan
title_sort n deglycosylation targeting chimera dglytac a strategy for immune checkpoint proteins inactivation by specifically removing n glycan
url https://doi.org/10.1038/s41392-025-02219-6
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