Amphiphilic Lipid-Single-Stranded DNA Conjugate-Mediated Cell Surface Engineering for Programmable Intercellular...

Intercellular tethering and interface stability critically influence cellular activation, particularly in solid tumors where physical constraints limit sustained effector-target engagement. In particular, effective immune-synapse formation in natural killer cells requires stable cell-cell contact. However, most existing strategies rely on tumor-antigen-mediated recognition and are therefore vulnerable to antigen heterogeneity and immune escape. Here, we developed an amphiphilic single-stranded DNA (ssDNA)-based surface-engineering strategy that enables controllable and receptor-independent regulation of intercellular interfaces. Lipid-conjugated ssDNA constructs were designed to (a) anchor onto cell membranes, (b) induce sequence-specific association through DNA hybridization, and (c) enable thermally reversible dissociation of tethered cell pairs. This membrane modification was rapidly achieved, and complementary ssDNA pairing markedly increased effector-target tethering, cytotoxic granule and cytokine secretion, and elimination of triple-negative breast cancer cells. Importantly, this platform remained effective in 3-dimensional tumoroid models, where amphiphilic ssDNA enabled robust membrane localization and facilitated natural-killer-cell-mediated tumor disruption. Collectively, these results demonstrate that immune-synapse efficiency could be actively modulated by engineering the physical properties of intercellular interfaces. Moreover, this programmable ssDNA-based platform offers a versatile framework for regulating diverse cell-cell interfaces, with broad applicability across immunotherapy, tissue engineering, and cell-based therapeutic systems.