Extended Data Fig. 9: CREM KO improves CAR-NK cell function in multiple in vivo models.
From: CREM is a regulatory checkpoint of CAR and IL-15 signalling in NK cells
(a) Body weight of the mice over time in the various groups in experiment depicted in Fig. 4a (n = 5 mice/group); (b) Ex vivo cytotoxicity of CREM WT and CREM KO CAR70/IL-15 NK cells harvested from mice sacrificed at day 10 and day 20 following NK cell infusion compared to the infusion products against mCherry+ K562 (n = 5 mice in each group); (c) Representative images from the assay in (b) at day 10; (d) Ex vivo cytotoxicity of CREM WT and CREM KO CAR70/IL-15 NK cells harvested from mice sacrificed at day 10 and day 20 following infusion compared to the infusion products against GFP+ Raji (n = 5 mice in each group); (e) Representative images from the assay in (d) at day 10; (f) Heatmap for the concentrations of the various cytokines in the serum of mice from day 10 and day 20 post-infusion in pg/ml; (g) Representative H&E staining of full lung sections from mice injected intravenously with BCX.010 PDX cell line and treated with CREM WT NT, CREM WT CAR70/IL-15, or CREM KO CAR70/IL-15 NK cells from the timed sacrifice experiment depicted in Fig. 4g; (h) Body weight of the mice over time in the various groups of the survival experiment depicted in Fig. 4g; (i) Quantification of flow cytometry analysis of the number of hCD45 + CD56 + CD16 + CAR+ (CD27 + ) cells (a marker of CAR+ NK cell engraftment) in the blood of mice 10 or 20 days after NK cell infusion in the various groups; data in (h) and (i) were pooled from two CB donors (n = 4 mice in BCX.010 alone group and n = 10 mice in each of the NT, CREM WT CAR70/IL-15 and CREM KO CAR70/IL-15 groups with 5 mice/CB donor); (j) Schematic illustration of the in vivo experimental plan for the PATC148 orthotopic mouse model. Created in BioRender. Rafei, H. (2025) https://BioRender.com/ryvr1mz; (k) Number of pancreatic tumor nodules in mice that were treated with CREM WT or KO CAR.TROP2/IL-15 NK cells (n = 4-5 mice per group); (l) Graph depicting flow cytometry analysis of the number of hCD45+ CD56+ CD16+ cells (a marker of NK cell engraftment) in the blood of mice 10 or 20 days after NK cell infusion in the CREM WT or KO CAR.TROP2/IL-15 NK cell groups (n = 4-5 mice per group). ns: non-significant. Statistical comparisons were performed using two-way ANOVA with Tukey correction (a,b-left,d-left,h,i), two-way ANOVA (b-right,d-right); two-tailed t-test (k), and two-way ANOVA (Fisher’s LSD test, l). Data are represented as mean ± SEM.
