Adoptive cell therapies(ACTs)have achieved remarkable clinical success in treating cancers;however,their broader application is greatly impeded by high cost and restricted antigen specificity.Recently,engineering the ...Adoptive cell therapies(ACTs)have achieved remarkable clinical success in treating cancers;however,their broader application is greatly impeded by high cost and restricted antigen specificity.Recently,engineering the glycocalyx has provided a convenient transgene-free means to design ACTs with highavidity glycan ligands to target CD22,offering a new avenue for B lymphoma immunotherapy.In this work,we perform a comparative analysis of the molecular profiles involved in metabolic or chemoenzymatic glycocalyx engineering and explore their multiplexing capability.The glycoproteomic results revealed content-dependent customization of the natural killer(NK)-92MI glycocalyx.Compared with metabolic engineering,exogenous chemoenzymatic engineering has comparable or even superior ligand-loading efficiency,with some immune synapse components modified to facilitate their spatial recognition against target cells.Next,we tested the orthogonal creation of ligands on NK-92MI cells by further engineering a2,3-sialylated N-acetyllactosamine moieties to produce selectin ligands that are essential for better in vivo eradication of mouse xenograft B lymphoma.Finally,we demonstrate that analogous engineering of CD19-targeted chimeric antigen receptor T(CAR-T)cells to produce CD19/CD22 bitargeted therapy can enhance antigen targeting and tumor cell killing,offering an alternative cost-efficient agent for treating cancer relapse with decreased levels of CD19 antigens.These findings establish a mechanistic foundation for glycocalyx engineering and support the rational design of nextgeneration ACTs against B lymphoma.展开更多
基金financial support from the National Natural Science Foundation of China(32150027 and 22177002)the National Key Research and Development Program of China(2022YFC3400803 and 2023YFC2308003)supported by the Russian Science Foundation(5-74-30002)。
文摘Adoptive cell therapies(ACTs)have achieved remarkable clinical success in treating cancers;however,their broader application is greatly impeded by high cost and restricted antigen specificity.Recently,engineering the glycocalyx has provided a convenient transgene-free means to design ACTs with highavidity glycan ligands to target CD22,offering a new avenue for B lymphoma immunotherapy.In this work,we perform a comparative analysis of the molecular profiles involved in metabolic or chemoenzymatic glycocalyx engineering and explore their multiplexing capability.The glycoproteomic results revealed content-dependent customization of the natural killer(NK)-92MI glycocalyx.Compared with metabolic engineering,exogenous chemoenzymatic engineering has comparable or even superior ligand-loading efficiency,with some immune synapse components modified to facilitate their spatial recognition against target cells.Next,we tested the orthogonal creation of ligands on NK-92MI cells by further engineering a2,3-sialylated N-acetyllactosamine moieties to produce selectin ligands that are essential for better in vivo eradication of mouse xenograft B lymphoma.Finally,we demonstrate that analogous engineering of CD19-targeted chimeric antigen receptor T(CAR-T)cells to produce CD19/CD22 bitargeted therapy can enhance antigen targeting and tumor cell killing,offering an alternative cost-efficient agent for treating cancer relapse with decreased levels of CD19 antigens.These findings establish a mechanistic foundation for glycocalyx engineering and support the rational design of nextgeneration ACTs against B lymphoma.
