Tumour priming by ultrasound mechanogenetics for CAR T therapy

Abstract Cell-based cancer immunotherapy holds potential as a therapeutic approach, yet its application for solid tumour treatment remains challenging. Here we report a focused-ultrasound-based approach that mechanically induces the localized expression of CD19 antigen within a subpopulation of cells within solid tumours, which function as local ‘training centres’ to activate chimeric antigen receptor T cells. Activated chimeric antigen receptor T cells attack the whole cancer cell population near the tumour site, thus achieving cancer suppression. The system achieves targeted gene expression by integrating focused-ultrasound-triggered mechanical stimulation and the subsequent calcium response of cancer cells with a doxycycline-gated AND-logic genetic circuit, both of which need to be active for effective induction of CD19 expression. We validate the functionality of the approach in vitro, in organoids and in vivo, achieving direct control of user-designed gene expressions through FUS-mediated mechanical stimulation without the need of any cofactor, demonstrating the approach’s potential as a versatile platform for precisely controllable immunotherapy. Overall, our combinatorial approach offers a focused-ultrasound-controlled remote and non-invasive priming of solid tumours for effective and safe chimeric antigen receptor T cell immunotherapy via the induced production of clinically validated antigens.