Fig. 5: Glyco-bridge with tandem HPAs show antitumor activity against pancreatic cancer patient-derived xenograft model.
From: Tuning CAR-T cells by targeting cancer-associated glycan in pancreatic cancer
a Schematic representation of CAR-T cells with a tandem HPA bridge, showing expanded glycan targeting and reduced bridge-mediated activation. b Schematic of mesothelin-targeting CAR T cells with tandem HPAs-based glyco-bridge. c Graphic representation of CAR and bridge constructs. d Flow cytometry analysis of mesothelin, HPA, MUC1, and Tn-MUC1 expression on Capan-2 and PDX-derived cells from of n = 3 independent experimental replicates. e Real-time cytotoxicity assay against indicated PDX-derived cells with indicated CAR T cells at 1:1-2:1 E:T ratio. Results are mean ± s.d. of n = 3 independent measurements. f Strength of interaction between PDX1294 and indicated 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 NSG mice were subcutaneously injected with 5 × 106 Capan2 C1GALT1 KO cells on day −14. On Day 0, 5 mice per group received 2 x 106 CAR-T cells with HPA bridge or HPA-directed CAR-T cell or Untransduced T cell (UTD). Tumor growth curve (h), body weight change (i) and survival percentage (j) for Capan2 C1GALT1 KO model using Donor #1. Results are mean ± s.e.m. of n = 5 mice. Tumor growth curve (k), body weight change (l) and survival percentage (m) for Capan2 C1GALT1 KO model using Donor #2. Results are mean ± s.e.m. of n = 5 mice. n Representative IHC images for human CD3 staining across different organs from Fig. 5g–m (n = 5 mice). Scale bar, 100 μm. o–r, NSG mice were subcutaneously injected with 5 × 106 PDX1294 cells on day −14. On Day 0, 5 mice per group received 2 × 106 CAR-T cells with HPA bridge or CD19-bridge or (UTD). Tumor growth curve (p), body weight change (q) and survival percentage (r) for PDX1294 cell model. Results are mean ± s.e.m. of n = 5 mice. In e, f, h, k, p, and r statistical analysis was performed by two-way ANOVA with correction for multiple comparisons.
