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, Ac 4 ManNAda, 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.