Fig. 4: Snail lectin HPA-based CAR T cells show antitumor activity and associated toxicity against Tn antigen-positive pancreatic tumor cell line.
From: Tuning CAR-T cells by targeting cancer-associated glycan in pancreatic cancer
a Biosynthetic pathway for Tn antigen. b HPA staining quantified by H-score in PDAC versus normal pancreatic tissues (PA807; n = 31 and 7). H-score represents the weighted sum of cells with low (0.14–0.4), moderate (0.4–0.6), or strong (>0.6 OD) intensity. Significance was assessed by unpaired two-tailed t-test. c Representative IHC images of HPA-stained tissue sections from b. Scale bar, 100 μm. d Surface level of HPA binding on Capan-2 following itraconazole incubation at the indicated concentrations for 48 h. e Schematic showing HPA-based CAR T cells. f Strength of interaction between target cells and indicated HPA CAR-T cells. Percentage of total CAR-T cells remaining bound to target cells as the acoustic force ramp is applied from 0 to 1000 pN are shown. Results are mean ± s.e.m. of n = 3 independent measurements. g Percentage of CAR-T cells remaining bound to the target cells at the avidity plateau under 1000 pN force from Fig. 4f. Significance was assessed by unpaired two-tailed t-test. h Representative confocal images of HPA-stained cells. Scale bar, 10 μm. i Surface level of HPA binding on indicated target cells. j HPA binding on each cell from (g). Results are mean ± s.d. of n = 3 independent measurements. Real-time cytotoxicity assay against PDX1294 (k, l), Capan-2 (m), Capan2- C1GALT1 KO (n) target cells with indicated CAR T cells at 1:1-2:1 E:T ratio. Results are mean ± s.d. of n = 3 independent measurements. o NSG mice implanted s.c. with 5 × 106 Capan-2 C1GALT1 KO cells received 2 × 106 dual-HPA CAR-T or UTD cells on day 0 (5 mice/group; n = 3 donors). Body weight change (left) and survival percentage for n = 5 mice. p Representative HPA staining of tumor and major organs from o (n = 5 mice). Scale bar, 100 μm. In (d, i) representative flow cytometry data from three independent experiments. In f and k–n statistical analysis was performed by two-way ANOVA with correction for multiple comparisons. In (j), one-way ANOVA with Tukey’s post hoc tests.
