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

Sonogenetic holds great potential for modulating cellular functions and behaviors. However, developing hypersensitive ultrasound (US)-responsive receptors remains a substantial challenge. Here, we combine AlphaFold3 prediction with molecular dynamics simulations to identify a hypersensitive Gpr114 protein variant (T-Gpr114) that responds to short-pulse US stimulation for activation. We further design sonogenetic macrophages (MΦ) by fusing T-Gpr114 to a synthetic genetic circuit, this circuit initiates expression of the transcription factor ID3 upon short-pulse US treatment. In vivo assays using female tumor-bearing mice demonstrate that sonogenetic-MΦ suppress tumor growth with no obvious systemic toxicity, and exhibit enhanced anti-tumor efficacy relative to cells with constitutive ID3 expression. Single-cell RNA sequencing reveals that sonogentic-MΦ potentiate the recruitment recruitment of CD8+ T cells compared with constitutive ID3-overexpressing macrophages. In this work, this T-Gpr114-based US-hypersensitive gene circuit extends the scope of remote control over the activation of diverse cell types, thereby facilitating the programming of therapeutic cells. Sonogenetics holds great potential for modulating cellular functions and behaviours. Here the authors present a T-Gpr114 sonogenetic system for non-invasive cellular modulation and engineer T-Gpr114 -based macrophages for anti-tumour activity and enhanced CD8+ T cell recruitment.