Artificial Host-Guest Recognition Directs Glycometabolically Engineered Macrophages to Tumors

CD3ζ-based chimeric antigen receptor macrophage (CAR-M) therapy for solid tumors is limited by complex viral-mediated genetic engineering and the challenge of maintaining a durable pro-inflammatory phenotype within the immunosuppressive tumor microenvironment. Here, we introduce a supramolecular glycoengineering strategy that bypasses genetic modification. By utilizing a novel metabolic labeling agent, Ac₄ManNAda, we successfully install adamantane (Ada, a guest molecule) tags onto macrophage surfaces via native biosynthetic pathways to generate glycoengineered supramolecular macrophages (GSAR-M). This labeling not only significantly enhances recognition of β-cyclodextrin (β-CD, a host molecule)-tagged tumor cells via CD-Ada host-guest interactions but also, unexpectedly, acts as an intrinsic activator. It induces a sustained distinct activated state in GSAR-M, characterized by upregulated activation markers and enhanced migratory, phagocytic, and tumoricidal capacities. Furthermore, subsequent LPS stimulation of these cells (termed GSAR-M+) cooperatively amplifies F-actin content and pseudopodia formation, leading to superior tumor cell capture in vitro. In a murine 4T1 breast cancer model, this supramolecular glycoengineering strategy achieves profound tumor growth arrest and effectively remodels the immunosuppressive tumor microenvironment. This study establishes a streamlined, cost-effective, and non-viral engineering paradigm that integrates host-guest recognition with glycometabolic engineering, providing critical insights for the development of next-generation engineered immune cells in adoptive cell therapies.