Extended Data Fig. 10: CAR T cell generation and GPR183-dependent in vivo infiltration and in vivo efficacy.
From: Engineering NK and T cells with metabolite-sensing receptors to target solid tumors
a, Surface expression of anti-EpCAM-CAR was assessed by flow cytometry. GFP+CAR+ T cells were sorted (left). CAR T cells were transduced with either control or GPR183-ORF vectors and surface GPR183 expression was assessed by flow cytometry (right). b, A gating strategy for in vivo CAR T cell experiments. Only live (7-AAD negative) cells were considered. mCD45−hCD45+hCD8+ cells were identified as injected CAR T cells. IFNg and CD107a expression were further assessed at baseline and upon PMA/ionomycin stimulation. c, GPR183 increases CAR T cell tumor infiltration (biological replicate of the experiment shown in Fig. 6f with T cells derived from another human donor). Flow cytometry-based quantification of in vivo accumulation of CAR T cells in lung, spleen, and tumor. d, IFNγ and CD107a expression in tumor-infiltrating CAR T cells. (c-d) Data are presented as mean values ± SEM of GPR183OE-CAR T cells (n = 6 mice) and control-CAR T (n = 5 mice) compared via two-tailed unpaired Student’s t-tests. e, GPR183 increases CAR T cell in vivo efficacy (biological replicate of the experiment shown in Fig. 6h,i with T cells derived from another human donor). Growth and survival curve of EpCAMOE MDA-MD-231 breast cancer tumors in NSG mice treated with anti-EpCAM-CAR T (n = 6 mice), or GPR183OE anti-EpCAM-CAR T (n = 7 mice). In (e) data are shown as the mean ± SEM (left) and individual growth curves per mouse (middle). Data analysis was performed using two-way ANOVA with Sidak correction for multiple hypotheses testing for growth curve and log-rank test for survival curve.
