New research published in engineering We provide mechanistic insights into the glycoengineering of living cells and support the rational design of next-generation adoptive cell therapy (ACT) for B lymphoma, addressing limitations such as high cost and limited antigen specificity in current treatments. Researchers from Peking University and international organizations compared β-galactoside alpha 2,6 sialyltransferase 1 (ST6Gal1)-mediated metabolic glycoengineering (MGE) and chemoenzymatic glycoengineering (CeGE) in NK-92MI cells, focusing on their molecular profiles and multiplexing capabilities.
Glycoproteomic analysis revealed content-dependent customization of the NK-92MI glycocalyx. Exogenous ST6Gal1-assisted CeGE showed equivalent or higher ligand transduction efficiency than MGE, with modifications to specific immune synapse components that may promote spatial recognition toward target cells. The research team further investigated creating orthogonal ligands on NK-92MI cells by manipulating the α2,3-sialylated N-acetyllactosamine moiety to generate selectin ligands. Selectin ligands are important for improving in vivo clearance of murine xenograft B lymphomas.
In this study, we applied a similar manipulation to CD19-targeted chimeric antigen receptor T (CAR-T) cells to develop a CD19/CD22 dual-targeted therapy. Incorporation of 9-Nm-phenoxybenzamide-N-acetylneuraminic acid (MPBNeu5Ac) enhances antigen targeting and tumor cell killing, providing a cost-effective candidate for cancers with reduced CD19 antigen levels leading to recurrence. Both CeGE and MGE strategies maintain biocompatibility without compromising cell viability or proliferation, supporting their potential for clinical application.
These findings establish the mechanistic basis for glycoengineering and demonstrate its versatility as a transgene-free approach to enhance immune cell function. This non-genetically engineered approach expands the toolbox for optimizing ACT by enabling customizable loading of high avidity glycan ligands targeting CD22 and selectins, with implications for improved outcomes in B lymphoma and potentially other hematological malignancies. This study focuses on glycocalyx remodeling as a compatible and flexible strategy to impart customized targeting and transport properties to cellular immunotherapies and support the continued development of next-generation cell-based cancer therapies.
The paper “Comparative mechanistic study of live cell glycoengineering: improving adoptive cell therapy for B lymphoma” is authored by Yuxin Li, Tao Gao, Zhaoxin Han, Valeria M. Stepanova, Han Wang, Hongmin Chen, Alexey Stepanov, and Senlian Hon.
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Reference magazines:
Lee, Y. Others. (2025). A comparative mechanistic study of live cell glycoengineering: Improving adoptive cell therapy for B lymphoma. engineering. DOI: 10.1016/j.eng.2025.08.037. https://www.sciencedirect.com/science/article/pii/S2095809925005557?via%3Dihub
