Truncated Gpr114-based ultrasound-hypersensitive gene circuit for controlled expression of therapeutics

Sonogenetics possesses significant potential for modulating cellular functions and behaviors. However, developing hypersensitive ultrasound (US)-responsive receptors presents a substantial challenge. Here, we utilized AlphaFold3 and molecular dynamics simulation technology to identify a hypersensitive Gpr114 protein variant (T-Gpr114) that can be activated by short-pulse US stimulation. We then engineered sonogentic macrophages (MΦ) using T-Gpr114 and integrated with an engineered genetic circuit, enabling high-performance expression of the nuclear factor ID3 upon short-pulse US stimulation. In mice, sonogentic-MΦ suppressed tumor growth without causing obvious toxicity and outperformed their constitutively expressing counterparts. Single-cell RNA sequencing revealed that engineered sonogentic-MΦ exhibit enhanced phagocytosis and improved recruitment of CD8+ T cells compared to cells with constitutive ID3 expression. This T-Gpr114-based US-hypersensitive gene circuit can be extended to remotely control the activation of various cell types, thereby facilitating the programming of therapeutic cells and enhancing their suitability for clinical translation.