Journal: Nature Communications
Article Title: Multiplex gene-editing strategy to engineer allogeneic EGFR-targeting CAR T-cells with improved efficacy against solid tumors
doi: 10.1038/s41467-025-66737-1
Figure Lengend Snippet: a Flow cytometry staining of H226 tumor cells in vitro for CD73 expression. Isotype control (gray), anti-CD73 antibody (pink). b Immunofluorescent micrographs of H226 tumors resected from NCG mice 46 days post-implantation. Nucleated cells (DAPI, blue), hypoxia (Hypoxyprobe, green) and CD73 (pink). Representative images from four individual tumors from 10 to 20 different cutting surfaces. c Quantification of hypoxia in various tumor regions within resected H226 tumors from NCG mice determined by mean fluorescence intensity (MFI) of Hypoxyprobe. Representative image of a resected tumor section; quantification was performed across 6 independent slides (3 tumors per slide from individual mice) with an average of 13.5 regions of interest (ROI) analyzed per slide. d Spatial transcriptomics gene expression analysis from hypoxic regions in ( c ) (white = low hypoxia, light green = medium hypoxia, dark green = high hypoxia). Boxplots show the median (line), interquartile range (box), and whiskers extending to values within 1.5× the IQR. e 2 × 10 6 UTD T cells (white, n = 5 individual mice) or unedited (gray, n = 5 individual mice) and A 2A R-KO (red, n = 5 individual mice) CAR T-cells injected I.V. into H226 tumor-bearing NCG mice. Group average of tumor volumes measured via calipers over time (Two-sided Mann–Whitney t-test, n = group average of individual mice, mean ± SEM, P** = 0.0079, P** = 0.0072). f Cumulative tumor burden, calculated as area under the curve, from ( e ) (Two-sided Mann–Whitney t-test, n = average of individual mice as above, mean ± SD, n.s. = 0.0556, P** = 0.00379). g 2 × 10 6 UTD T cells (white, n = 5 individual mice) or unedited (gray, n = 5 individual mice) and A 2A R-KO (red, n = 5 individual mice) CAR T-cells injected I.V. into A549 tumor-bearing NCG mice. Group average of tumor volumes measured via calipers over time (Graph represents group mean ± SD, P** = 0.0072). h Cumulative tumor burden, calculated as area under the curve, from ( g ). (Two-sided Mann–Whitney t-test, n = average of individual mice as above, mean ± SD, n.s. = 0.490, P** = 0.0037). For all data, symbols and error bars reflect individual biological replicates and group mean ± S.E.M. e – h Mann–Whitney t-test performed to calculate statistical significance, ** P < 0.01, * P < 0.05.
Article Snippet: Representative image of a resected tumor section; quantification was performed across 6 independent slides (3 tumors per slide from individual mice) with an average of 13.5 regions of interest (ROI) analyzed per slide. d Spatial transcriptomics gene expression analysis from hypoxic regions in ( c ) (white = low hypoxia, light green = medium hypoxia, dark green = high hypoxia).
Techniques: Flow Cytometry, Staining, In Vitro, Expressing, Control, Fluorescence, Gene Expression, Injection, MANN-WHITNEY
