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. 2022 Aug 4;13:4443. doi: 10.1038/s41467-022-32198-z

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© The Author(s) 2022

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a Heatmap depicting the 1355 most differential expressed genes between the 3 CMS4-PM subgroups (1 PM sample per patient, n = 45), clustered by CMS4-PM subgroup. Location of lesions, primary tumor type, development of PM (syn- or metachronous), mutational status of CRC driver genes and MSI are indicated below (two-sided Fisher’s exact test). b Z-scores of KRAS (left) and TP53 (right) activation gene signatures in the CMS4-PM subgroups. Red and green dots indicate respectively mutated or wild-type genes (one-way ANOVA with Tukey’s multiple comparisons test, ***P = 0.0001, ****P < 0.0001, **P = 0.0049, *P = 0.0232). c Distribution of RAS mutations (KRAS and NRAS) over the 3 CMS4-PM subtypes. d Gene set enrichment analysis depicting differentially expressed (P < 0.05, two-sided t-test) signatures between the PM subgroups. Colors indicate relative signature expression (z-score). e Deconvolution of bulk RNA sequencing data was used to determine the fraction of immune cells per sample (Welch ANOVA with Dunnett’s multiple comparisons test, P = 0.0036, P = 0.0048, and P = 0.0166). f Relative distribution of immune cell types over total immune cell compartment (one-way ANOVA with Tukey’s multiple comparisons test, T cells *P = 0.049; Myeloid cells *P = 0.0476 and P = 0.0112; Mast cells *P = 0.0142). g Heatmap depicting relative expression of 18 immune checkpoint-associated genes within CMS4-PM subgroups. h Boxplots showing PCI scores within the 3 CMS4-PM subgroups (one-way Anova, P = 0.058). i Cumulative overall survival (OS) of patients with CMS4 classified peritoneal lesions, stratified to the 3 CMS4 subgroups. CMS4-PM.B has a significantly worse survival when compared to the other two combined (Breslow, P = 0.006). b, e, f, h n = 15 (CMS4-PM.A), n = 7 (CMS4-PM.B) or n = 23 (CMS4-PM.C) biologically independent samples. a, b, e, f, i Asterisks indicate level of significance: *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. b, e, f, h Boxplots indicate median, first and third quartiles (Q1 and Q3), whiskers extend to the furthest values. Source data are provided as a Source data file.