Ultrasound priming gated by solid tumor hallmarks to guide CAR-T therapy.

CAR-T therapy is highly effective in hematologic malignancies but remains challenging in solid tumors. To address this challenge, here, we developed SHIFTERS (solid-tumor hallmark inducible, focused-ultrasound triggered enhanced reprogramming system), an AND-gated circuit that rewires tumor hallmarks into the clinically validated antigen CD19 under focused-ultrasound (FUS) control. SHIFTERS uses a split Gal4-VP64 transcription factor with one module driven by a hypoxia-responsive promoter and the other by a FUS-inducible heat-shock promoter, restricting CD19 induction to sonicated, hallmark-positive regions. This design enables robust, sustained local CD19 expression to activate and train CD19 synNotch CAR-T cells. In 3D spheroids and in vivo models, hypoxia-gated SHIFTERS combined with synNotch CAR-T cells achieved strong tumor suppression, supporting FUS-guided, localized activation. SHIFTERS is modular and can be adapted to tumor-type-specific promoters such as enAFP in hepatocellular carcinoma to enable both spatial and cell type-specific targeting. Together, SHIFTERS provides a modular, tumor-constrained, ultrasound-controllable platform to improve the precision and efficacy of CAR-T therapy for solid tumors.