Extended Data Fig. 2: Ferroptosis inhibitors promote CAR-T cell viability and functionality in vitro.
From: Iron-mediated ferroptosis impairs CAR-T cell function and antitumor efficacy
a-c, Expansion curves of CAR-T cells treated with ferrostatin-1 (Fer-1) (a), liproxstatin-1 (Lip-1) (b), or UAMC-3203 (UAM) (c) at indicated concentrations. Inhibitors were added every 48 h from day 6 post-CAR-T cell generation. d-i, Flow cytometric analyses of Tcm (central memory T cells, CD45RO+, CD62L+) and Tem (effector memory T cells, CD45RO+, CD62L−) subsets in CAR-T cells pretreated with Fer-1 (d, g), Lip-1 (e, h) or UAM (f, i) for 72 h starting at day 6. j-l, Flow cytometric analysis of T-cell differentiation based on CD45RA and CD27 expression in CAR-T cells (j) or in CD4+ and CD8+ subsets (k, l) pretreated with the indicated inhibitors. m, Expression of exhaustion markers (PD-1, TIM3 and LAG3) in CAR-T cells treated with different doses of Fer-1, Lip-1, and UAM: group A (1 μM, 100 nM, 5 nM), B (2 μM, 200 nM, 10 nM), and C (4 μM, 400 nM, 20 nM). n-p, Tumor growth in mice not treated with CAR-T cells. Bioluminescence kinetics (n), representative bioluminescence images (o) and Kaplan-Meier survival curves (p) of luciferase-expressing Nalm6 tumor-bearing mice treated with vehicle or the indicated inhibitors by intraperitoneal injection (n = 5 mice per group). Images were acquired at the indicated time points from the same animals. Data in a-c, g-i and k-m are shown as mean ± s.d. of 3 technical replicates from one representative donor. The experiment was repeated using one additional independent donor, with similar trends observed for panels a-m; the related quantification is available in the Source Data. Panels d-f and j show representative flow cytometry plots from one representative donor. No statistical analysis was performed for a-c, g-i and k-m. P values were calculated by two-way ANOVA with Dunnett’s multiple-comparisons test (n), log-rank (Mantel-Cox) test (p).
