Equipping Natural Killer Cells with Cetuximab through Metabolic Glycoengineering and Bioorthogonal Reaction for Targeted Treatment of KRAS Mutant Colorectal Cancer

Abstract

While Cetuximab can be used to treat KRAS wild-type colon cancer cells by targeting EGFR and inhibiting the activation of downstream signaling pathways, it exhibits little therapeutic effect on KRAS mutant colon cancer cells. Natural killer (NK) cells are a class of powerful immune cells with anticancer activities. However, NK cells typically lack inherent tumor targeting abilities. Here, a new method is established to bestow NK-92 cells with tumor targeting abilities by installing cetuximab on the cell surface. Through metabolic glycoengineering, azide groups were introduced onto the surface of NK-92 cells. Bioorthogonal strain promoted the azide–alkyne cycloaddition click reaction of engineered NK-92 cells with alkyne modified cetuximab functionalized NK cells with the antibody. The resulting NK-92 cells were significantly more effective than the parent NK-92 cells in protecting against tumor development in a KRAS mutant mouse tumor model resistant to cetuximab treatment. Thus, NK cell functionalization with antibodies enabled by metabolic glycoengineering is a promising strategy to enhance anticancer immune therapy.

General and Robust Chemoenzymatic Method for Glycan-Mediated Site-Specific Labeling and Conjugation of Antibodies: Facile Synthesis of Homogeneous Antibody–Drug Conjugates

“…Taken together, the results suggest that ADCs made by this method carrying 6–8 payloads per antibody might result in an optimal potency and safety profile. In addition to ADCs, the azido-tagged antibodies described here can be also used for other antibody conjugations, such as site-selective construction of antibody–enzyme conjugates, , antibody–cell conjugates, , antibody–antibiotic conjugates, and lysosome-targeting chimeras for targeted protein degradation. − …”

“…In addition to ADCs, the azido-tagged antibodies described here can be also used for other antibody conjugations, such as site-selective construction of antibody−enzyme conjugates, 73,74 antibody−cell conjugates, 75,76 antibody−antibiotic conjugates, 77 and lysosome-targeting chimeras for targeted protein degradation. 78−81…”

Metabolic glycoengineering – exploring glycosylation with bioorthogonal chemistry

“…209 Recently, NK cell surfaces were decorated with azide functionalities via MGE using 9-azido N-acetyl neuraminic acid methyl ester. 211 The tumor-targeting capacity was improved by bioconjugation of DBCO-modified cetuximab to the azide groups presented on the NK cell surface. Cetuximab is an antibody, which is specific for the epidermal growth factor receptor, and it is widely used in cancer immunotherapy.…”

KRAS Mutations in Solid Tumors: Characteristics, Current Therapeutic Strategy, and Potential Treatment Exploration

“…The inhibition of ERK signaling was found to assist in reducing PD-L1 expression through autophagy in intrahepatic cholangiocarcinoma (iCCA), indicating that ERK-targeted therapy may be combined with anti-PD-(L)1 immunotherapy for KRAS-mutant iCCA [ 135 ]. Equipping cetuximab on the surface of NK cells may solve the problem of cetuximab resistance in KRAS-mutant CRC [ 136 ].…”