A new study published in Engineering provides mechanistic insights into live-cell glycocalyx engineering and supports the rational design of next-generation adoptive cell therapies (ACTs) against B lymphoma, addressing limitations such as high costs and restricted antigen specificity in current treatments. Researchers from Peking University and international institutions compared metabolic glycocalyx engineering (MGE) and chemoenzymatic glycocalyx engineering (CeGE) mediated by β-galactoside α2,6 sialyltransferase 1 (ST6Gal1) 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 comparable or higher ligand-loading efficiency than MGE, with modifications to certain immune synapse components that may facilitate spatial recognition against target cells. The team further explored orthogonal ligand creation on NK-92MI cells by engineering α2,3-sialylated N-acetyllactosamine moieties to generate selectin ligands, which are critical for improved in vivo elimination of mouse xenograft B lymphoma.

The study also applied similar engineering to CD19-targeted chimeric antigen receptor T (CAR-T) cells to develop CD19/CD22 bitargeted therapy. Incorporation of 9-N-m-phenoxybenzamide–N-acetylneuraminic acid (^MPBNeu5Ac) enhanced antigen targeting and tumor cell killing, offering a cost-effective candidate for cancers with reduced CD19 antigen levels that lead to relapse. Both CeGE and MGE strategies maintained biocompatibility without impairing cell viability or proliferation, supporting their potential for clinical translation.

These findings establish a mechanistic foundation for glycocalyx engineering and demonstrate its versatility as a transgene-free approach to enhance immune cell functions. By enabling customizable loading of high-avidity glycan ligands targeting CD22 and selectins, this non-genetic engineering method expands the toolbox for optimizing ACTs, with implications for improving treatment outcomes in B lymphoma and potentially other hematologic malignancies. The work highlights glycocalyx remodeling as a compatible and flexible strategy to endow cellular immunotherapies with tailored targeting and trafficking properties, supporting continued development of next-generation cell-based cancer treatments.

The paper “A Comparative Mechanistic Study of Live-Cell Glycocalyx Engineering: Improving Adoptive Cell Therapies Against B Lymphoma,” is authored by Yuxin Li, Tao Gao, Zhaoxin Han, Valeria M. Stepanova, Han Wang, Hongmin Chen, Alexey Stepanov, Senlian Hong. Full text of the open access paper: https://doi.org/10.1016/j.eng.2025.08.037. For more information about Engineering, visit the website at https://www.sciencedirect.com/journal/engineering.