Fig. 5: Combinatorial immunotherapy in vitro.
From: Tumour priming by ultrasound mechanogenetics for CAR T therapy
a, Schematic diagram of the combinatorial immunotherapy strategy. First, tumours are transduced with the CaDox genetic system. Upon localized FUS stimulation, the CaDox system is activated in a subset of tumour cells, leading to ATP release via PANX1 hemichannels and calcium influx through purinergic receptors. This results in the expression of clinically validated antigens, such as tCD19, which prime synNotch CAR T cells (synNotch T). Activated synNotch CAR T cells then express anti-PSMA CAR to target and eliminate surrounding tumour cells. b, PC-3 cells were equipped with the CaDox–tCD19–EGFP system to express tCD19 and EGFP upon FUS activation. c,d, Representative images of induced gene expression (c) and quantified EGFP expression (d) levels measured in no treatment (n = 10) or combined treatment (n = 6) groups. Data represent mean ± s.e.m. Statistical significance was determined by unpaired two-tailed Welch’s t-test; adjusted P < 0.0001. e, Primary T cells were engineered with anti-CD19 (αCD19) synNotch and inducible anti-PSMA CAR to be activated by the induced tCD19+ cells to attack and clear PSMA+ cancer cells. UAS, upstream activating sequence (Gal4-binding promoter); SynNotch T, synthetic Notch T cells. f, Representative images of SYTOX staining to assess the cytotoxicity of PC-3 cells. Dashed line indicates the FUS-targeted region. g, Quantification of cytotoxicity of PC-3 cells in no treatment (n = 7) and treatment (n = 10) groups. Data represent mean ± s.e.m. Statistical significance was determined by unpaired two-tailed Welch’s t-test; adjusted P < 0.0001. Images in c and f are representative of ≥3 independent experiments. **** denotes adjusted P < 0.0001. Panels a, b and e created with BioRender.com.
