Figure 1. CAR T-cell therapy improves survival in Group 3 medulloblastoma orthotopic xenograft models.

(a) Experimental scheme - Patient-derived Group 3 medulloblastoma cells expressing eGFP-firefly luciferase (PDX-GFP-Luc) were xenografted into NSG mice. Bioluminescence imaging (BLI) was performed to determine orthotopic engraftment, at which point a single dose of 5×10⁶ (equivalent to a 20:1 ratio tumour cells:CAR-T-cells) monovalent EPHA2 CAR-T-cells, trivalent CAR-T (EPHA2, HER2 and IL13Rα2) cells, or non-transduced T-cell controls were injected into intraventricularly via the lateral ventricle. Tumour burden was monitored weekly by BLI until humane endpoint. (b) Survival analysis of T-cell treated xenografts of Med-114FH, Med411FH and MDT-MMB. Two-sided log-rank test Benjamini-Hochberg (BH), n = 19 EPHA2 CAR T-cells, 19 TRI CARs-T-cells, 19 non-transduced T-cells and 13 no treatment (NT) controls across 3 independent PDX models. Med114FH: EPHA2 vs T-cells P=0.004, EPHA2 vs NT P=0.0025, EPHA2 vs TRI P=0.9223, TRI vs T-cells P=0.0134, TRI vs NT P=0.0040; Med411FH: EPHA2 vs T-cells P=0.002, EPHA2 vs NT P=0.004, EPHA2 vs TRI P=0.2439, TRI vs T-cells P=0.0017, TRI vs NT P=0.0040; MDT-MMB: EPHA2 vs T-cells P=0.0062, EPHA2 vs NT P=0.0063, EPHA2 vs TRI P=0.0062, TRI vs T-cells P=0.2138, TRI vs NT P=0.2709 (c) BLI and final Hematoxylin-eosin (H&E) staining analysis of NSG mice xenografted with Med114FH, intraventricularly infused via the lateral ventricle with non-transduced T-cells, TRI CAR T-cells or EPHA2 CAR T-cells (colour map for all images indicates radiance), n = 7 EPHA2 CAR T-cells, 5 TRI CARs-T-cells, 7 non-transduced T-cells and 5 no treatment (NT) biologically independent PDX models. Each column represents one mouse, each row represents a time point, with mouse endpoint (days post therapy) noted in ‘Days’.