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. 2024 Nov 16;15:9949. doi: 10.1038/s41467-024-54364-1

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — a GO:BP term enrichments for genes with upregulated expression in the Club (top) and Immune (bottom) regions. One-sided Fisher’s exact test was used. b Correlation between Club region fraction and PMN-MDSC activity scores in pseudo-bulk for the discovery (left) and the validation (right) cohorts. Two-sided Spearman correlation coefficient and p-value are shown. Error bands in light grey span the 95% confidence interval calculated using a bootstrap. BPH: benign prostatic hyperplasia (n = 4), TRNA: treatment-naïve prostate cancer (n = 17); NEADT: neoadjuvant-treated prostate cancer (n = 22); CRPC: Castration-resistant prostate cancer (n = 5). Low cancer% n = 11, Mid cancer % n = 11, High cancer % n = 10. c PMN-MDSC activity score in treatment-naïve spots grouped according to their proximity to the Club region. Asterisks indicate two-sided independent samples t-test significance levels (p < 0.05) and effect size: *70th percentile <mean; **80th percentile <mean; ***90th percentile <mean. Club region (n = 278), proximal Tumor (n = 687), distant Tumor (n = 8751). d Violin plots of gene set activity scores across all regions in treatment-naïve (n = 36,198) and neoadjuvant-treated (n = 59,259) samples. Asterisks are the same as in (c). e A representative immunostained prostate tissue section (total n = 16) with selected regions of interest (ROIs). f Representative images of club-like positive (n = 47) and club-like negative (n = 54) ROIs corresponding to those annotated in (e). g A representative image of a club-like positive ROI (n = 47) with a three-colour stain showing CD66b⁺CD11b⁺CXCR2⁺ PMN-MDSCs. Arrows point to example cases. Scale bars in white are 500 μm in each panel. h Scatterplot of log-transformed club-like cell and PMN-MDSC cell counts. Each dot represents a ROI (n = 101). Correlation coefficient, p-value, and error bands were calculated the same as in (b). i Violinplot of PMN-MDSC percentage of all detected cells in club-like negative (n = 54) and club-like positive (n = 47) ROIs. A Wilcoxon rank-sum test p-value is shown.