Fig. 5: Prophylactic uPAR CAR T cells rejuvenate ISCs.
a, Experimental scheme for q–t: Young (3 months) mice were treated with 0.5 × 106 untransduced T cells (UT) or uPAR CAR T cells (m.uPAR-m.28z). Mice were harvested 15 months after infusion at the age of 18 months, and scRNA-seq was performed from whole small intestine: duodenum, jejunum and ileum (n = 1 per group). b, UMAP visualization of small intestinal cell types generated by 10X chromium protocol. Color scale indicates difference in localized cellular density between m.uPAR-m.28z and UT treated mice. c, Fraction of cells for each of the different cell types shown in q in mice treated with UT or m.uPAR-m.28z cells. d, UMAP visualization of small intestinal cell types generated by 10X chromium protocol. Color scale indicates log2FC differences in stemness signature score between m.uPAR-m.28z- and UT-treated mice. e, Split-violin plot indicates the expression level of five different stem-related genes in the stem cells from old UT- and m.uPAR-m.28z-treated mice. Boxplots display median (center line) and interquartile range (box). f, Experimental scheme for v–w: Young (3 months) mice were treated with 0.5 × 106 untransduced T cells (UT) or uPAR CAR T cells (m.uPAR-m.28z). Mice were harvested 15 months after infusion at the age of 18 months and organoids were generated from their intestinal crypts (n = 3 mice per group for UT and n = 4 mice for m.uPAR-m.28z, four replicates per mouse). g, Representative images of organoids at day 4. h, Number of organoids per field at day 4 (n = 3 mice per group for UT and n = 4 mice for m.uPAR-m.28z, four replicates per mouse). Shown are results of one independent experiment (a–h). Significance was determined by two-tailed Wilcoxon rank-sum test (*P < 0.05,**P < 0.01, ***P < 0.001, ****P < 0.0001) (e). Data are mean ± s.e.m. (h). Significance was determined by two-tailed unpaired Student’s t-test (h). Illustration was created with Biorender.com (a,f).
