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Chemo-enzymatic Synthesis of N-glycans for Array Development and HIV Antibody Profiling
Published on: February 5, 2018
N-deglycosylation targeting chimera (DGlyTAC): a strategy for immune checkpoint proteins inactivation by specifically
Li Li1, Jiajia Wu2, Weiqian Cao3
1Tongji University Cancer Center, Shanghai Tenth People's Hospital, School of Medicine, Tongji University, Shanghai, China.
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.
Insights
A novel deglycosylation targeting chimera (DGlyTAC) technique specifically removes N-glycans from target proteins like PD-L1. This approach shows promise for cancer immunotherapy by inactivating immune checkpoints, with superior in vivo therapeutic effects compared to antibodies.
Area of Science:
- Biotechnology
- Immunology
- Cancer Therapy
Background:
- Immune checkpoint inhibition is key for anti-cancer immunity.
- N-glycosylation is crucial for immune checkpoint protein function.
- Lack of specific de-N-glycosylation techniques limits therapeutic potential.
Purpose of the Study:
- To develop a novel, target-specific N-glycan removal technique.
- To assess the efficacy of this technique in cancer immunotherapy.
Main Methods:
- Developed deglycosylation targeting chimera (DGlyTAC) using Peptide-N-glycosidase F (PNGF) fused with target-specific nanobodies/affibodies.
- Applied DGlyTAC to target cell surface immune checkpoints (CD24, CD47, PD-L1).
- Evaluated specificity and off-target effects on overall N-glycosylation.
Main Results:
- DGlyTAC achieved specific N-glycan removal from target proteins with minimal impact on other proteins.
- Successfully inactivated immune checkpoints, particularly PD-L1.
- PD-L1 targeted DGlyTAC demonstrated superior in vivo therapeutic effects against tumors compared to PD-L1 antibodies.
Conclusions:
- Created a novel target-specific N-glycosylation erasing technique (DGlyTAC).
- DGlyTAC offers a modular strategy for inactivating membrane proteins.
- This technique has broad implications for developing advanced tumor immune therapeutics.
