Spatial mapping of NK cells. A, Percentage of CD8+ T-cell subsets and NK cells of all immune cells plotted among three tumor-infiltrating lymphocytes: “absent” (cold), “non-brisk” (excluded), and “brisk” (hot) assessed by pathologist grouping all samples and splitting by response for the NK cells. Data analyzed by a two-sided Wilcoxon test and bold = P B, Representative multiplex immunofluorescence staining on lesions from an NR for whom NK cells were detected in the immune-infiltrating patch. Green = GNLY, red = melanoma, orange = CD3, and blue = CD8. Arrows point NK cells, and the dashed line represents tumor border. C, Density per square millimeter of NK cells compared between three different tumoral regions, response, and time point. Data analyzed by a two-sided Wilcoxon test and bold = P D, Spatial plots of all identified cells from two examples of immune-excluded tumors from skin metastasis biopsies. Arrows point toward NK cells, and the dashed line represents tumor border. E, Dotplot of expression of NK-related genes by NK cells plotted among immunophenotype and time point; BT cold, n = 1; BT excluded, n = 1; OT cold, n = 1; and OT excluded, n = 5; (F) Heatmaps of neighborhood enrichment of cell types/states identified using Xenium spatial transcriptomics for one BT excluded sample and five OT excluded samples. Cells for which abundance was z-scores from a permutation test, indicating how frequently each pair of cell types was observed as neighbors compared with a randomly permuted spatial distribution. G, Stacked bar representing percentages of 20 immune cells surrounding NK cells, plotted between one BT excluded sample and five OT excluded samples. CAF, cancer-associated fibroblast; cDC1, conventional type 1 DCs; cDC2, conventional type 2 DCs; R, responder; Treg, regulatory T cell.
Poźniak et al. (Thu,) studied this question.