Fig. 6: Microglia impact on the GD2 CAR T cell response.
a, Representative immunofluorescence 3D images of a 200-µm-thick slice containing microglia, 1 week after start of GD2 CAR T cell treatment. Cells are labeled for DAPI (white), GFP+ DMG tumor cells (green), IBA1+ microglia (orange), CD3+ T cells (cyan) and cCasp3 (red). n = 2 DMGOs. Bottom, zoomed-in view of area in white inset (n = 2 DMGOs). Scale bar, 10 µm. b, Heat map depicting the fold change in concentration of selected cytokines, chemokines and growth factors of DMGOs containing microglia normalized against no microglia on day 3, day 7 and day 14 after GD2 CAR T cell addition (n = 6 DMGOs). TNF, tumor necrosis factor. c, Percentage of cells within GD2 CAR T cell clusters, including a new microglia-affected cluster, for nonexposed GD2 CAR T cells (left), GD2 CAR T cells retrieved from DMGO without (middle) or with integrated microglia (right). d, Heat map highlighting the average scaled expression of curated TIL gene signatures46 in the TMA GD2 CAR T cell cluster. e, Reduction in tumor area (normalized z score per DMGO relative to time point 0) after the addition of GD2 CAR T cells in DMGO without (blue) or with (orange) integrated microglia. The arrow indicates the time point of GD2 CAR T cell administration. Data are shown as the mean ± s.e.m. Statistical analysis at each time point was performed using a linear mixed-effects model, accounting for experimental and organoid variation. Reported P values are two-sided and were adjusted for multiple comparisons using the false discovery rate (Benjamini–Hochberg) (t3, P = 0.04389; t7, P = 0.04389; t10, P = 0.08409; t14, P = 0.0578). For b–e, n = 6 DMGOs with microglia and n = 6 DMGOs without microglia from four independent batches.
