Fig. 5: TAK1 inhibition does not cause a pro-inflammatory immune response.
a Experimental approach to assess the immune response elicited by ubiquitous TAK1 inhibition on tumor spheroids generated by isolation of total cell populations from tumor tissue of one PDAC-patient71. b Cell viability assay of PDAC-derived tumor spheroids (n = 4 per condition) treated with DMSO or 5Z-7-Oxozeaenol (25 µM) for 72 h. Results are expressed as mean ± SEM. P value was calculated by Mann–Whitney U test (two-tailed), *p = 0.0286. c UMAP of single-cell transcriptomes showing DMSO and 5Z-7-Oxozeaenol-treated immune cells (DMSO: n = 208; 5Z-7-Oxozeaenol: n = 78). d–f UMAP of CD8+ (DMSO: n = 67; 5Z-7-Oxozeaenol: n = 7), CD3E+ (DMSO: n = 117; 5Z-7-Oxozeaenol: n = 22) and CD44+ (DMSO: n = 122; 5Z-7-Oxozeaenol: n = 23) T-cells, isolated from PDAC-derived tumor spheroids, indicating T-cells as the main cell population. Color bar indicates log2-normalized expression. g, h Gene Set Enrichment Analysis (GSEA) in cluster 1 of T-cells isolated from DMSO vs. 5Z-7-Oxozeaenol-treated PDAC patient-derived tumor spheroids. Normalized enrichment score (NES) of significantly enriched (red), suppressed (blue) or non-significantly regulated (gray) (FDR > 0.05) pathways after 5Z-7-Oxozeaenol treatment (***FDR q < 0.001) are presented. i Experimental approach to assess the immune response elicited upon TAK1 inhibition on CD45+ cell-depleted tumor spheroids generated by isolation and in vitro reconstitution from tumor tissue of one PDAC-patient72. j Cell viability assay of PDAC-derived tumor spheroids (n = 6 per condition) treated with DMSO or 5Z-7-Oxozeaenol (1 µM, 10 µM and 25 µM) for 72 h. Results are expressed as mean ± SEM. P value was calculated by ordinary one-way ANOVA (two-tailed) with Tukey’s multiple-comparisons test with n.s. = not significant, *p = 0.0114, ****p < 0.0001. k UMAP of single-cell transcriptomes of PDAC-derived immune cells subjected with the supernatant of DMSO- or 5Z-7-Oxozeaenol (25 µM)-treated tumor spheroids (DMSO: n = 529; 5Z-7-Oxozeaenol: n = 427). l GSEA of PDAC-derived T-cells incubated with the conditioned medium from DMSO- or 5Z-7-Oxozeaenol-treated tumor spheroids. NES was non-significantly altered (FDR > 0.05) in all pathways that were affected after 5Z-7-Oxozeaenol treatment in g-h. Source data are provided in the Source Data file.
