An antibody‐drug conjugate targeting a GSTA glycosite‐signature epitope of MUC1 expressed by non‐small cell lung cancer

Antibodies targeting aberrantly glycosylated proteins are ineffective in treating cancer. Antibody‐drug conjugates have emerged as effective alternatives, facilitating tumor‐specific drug delivery. Previous studies have assessed the aberrantly glycosylated tandem repeat region of MUC1 glycoprotein as three site‐specific glycosylated neoantigen peptide motifs (PDTR, GSTA, and GVTS) for binding with a monoclonal antibody. This study aimed to develop an antibody‐drug conjugate for cancer treatment based on monoclonal antibodies against the aforementioned three neoantigen peptide motifs. Internalization of monoclonal antibodies was assessed via immunofluorescence staining and colocalization with lysosomal markers in live cells. Antibody positivity in tumor and peritumoral tissue samples was assessed via immunohistochemistry. The efficacy of anti‐MUC1 ADCs was evaluated using various cancer cell lines and a mouse tumor xenograft model. An anti‐MUC1 ADC was synthesized by conjugating GSTA neoantigen‐specific 16A with monomethyl auristatin E (MMAE), which displayed potent antitumoral efficacy with an IC50 ranging 0.2–49.4 nM toward various cancer cells. In vivo, 16A‐MMAE inhibited tumor growth in a dose‐dependent manner in a mouse xenograft model established using the NCI‐H838 NSCLC cell line, at a minimum effective dose of 1 mg/kg. At 3 mg/kg, 16A‐MMAE did not cause significant toxicity in a transgenic mouse expressing human MUC1. The high antitumoral efficacy of 16A‐MMAE suggests that aberrant glycosylated MUC1 neoantigen is a potential target for the development of ADCs for treating various cancers. Personalized therapy may be achieved through such glycosite‐specific ADCs. In this study, we describe an anti‐MUC1 antibody‐drug conjugate, 16A‐monomethyl auristatin E (MMAE). 16A‐MMAE showed potent antitumoral activity in multiple types of cancer, suggesting that neoantigens generated by posttranslational modification are promising targets for antibody‐drug development.