Targeting hypersialylation in pancreatic cancer microfluidic models reverses its malignant phenotype and enhances CAR-T therapy

Pancreatic ductal adenocarcinoma (PDA) exhibits extensive hypersialylation, a glycosylation hallmark associated with tumor progression and immune evasion. Here, we employed physiologically relevant 3D microfluidic models, which mimic key aspects of the tumor microenvironment, to evaluate the impact of sialyltransferase inhibition using the sialic acid analog Ac 5 3F ax Neu5Ac in PDA cell models. This glycomimetic compound reduced α2,3-sialylation without compromising cell viability in self-organized PDA spheroids. Desialylation decreased the number and velocity of migrating cells in confined and unconfined microenvironments, and reduced cell adhesion to engineered endothelial vessels, consistent with reduced metastatic ability. Notably, 3D co-culture with CAR-T cells revealed enhanced T cell-mediated cytotoxicity upon sialylation inhibition, leading to reduced spheroid size and viability. Collectively, this work provides new insights into the role of sialylation in modulating tumor morphology, cell viability, migratory behavior, and interactions with immune effector cells, underscoring the value of biomimetic microfluidic models for evaluation of anti-metastatic strategies. Overall, targeting hypersialylation reprograms the malignant phenotype of PDA and may enhance immunotherapeutic efficacy, supporting the development of glycan-targeting strategies in cancer therapy.