Fig. 1: Kinase inhibition induces senescence-associated inflammatory signaling.

a Core gene set of a combined gene set enrichment analysis (GSEA) in oncogene-driven cancer cell lines (PC9, HCC827, HCC4006, H1993, H3122, A375, Colo205) after 3 days of treatment with their respective kinase inhibitor. b Cell count of EGFR^mut PC9 cells under osimertinib (osi, 300 nM). The upper line indicates normalized cell number, shaded areas corresponding cell cycle distribution (n = 3). Inset: Immunoblot of cell cycle regulator genes in PC9 cells after 5 days of osimertinib treatment. c, d GSEA of time-series RNAseq in EGFR^mut cells indicates temporal adaptation processes (ATSC = adult tissue stem cell gene set, NES = Normalized enrichment score). e GSEA of the ATSC and the IFNα gene set across the two EGFR^mut PDX models (osi vs. vehicle). f Schematic of the humanized mouse model (top) and exemplary histology of low (−), medium (+), and high (++) CD8 T cell infiltration (bottom). Scale bar 100 µm, representative images of in total n = 19 tumors of n = 10 mice. g Digital pathology-based quantification of T cell infiltration in humanized mice following 4 days of treatment with osimertinib (5 mg/kg, n = 9 tumors) or vehicle (n = 10 tumors) (error bars indicate mean ± SEM). h Hyperion imaging mass cytometry false-color image of an osimertinib treated tumor from (f) stained for pan-cytokeratin (CK, cyan), CD8 (green), and Granzyme B (red). The overlay of CD8 and red is colored yellow. Scale bar 100 µm, representative image of n = 6 regions. i Flow cytometry analysis of infiltrating T cells in humanized PC9 xenografts after 4 days of treatment (in total n = 6 tumors of n = 3 mice per group; error bars indicate mean ± SEM). j RNA-seq-based GSEA of public BRAF^mut melanoma patient data, comparing patients before (Pre-treatment) with patients during BRAF or BRAF + MEK inhibition (On-treatment). k Proportion of CD8 T cell infiltration inferred from bulk RNA-seq in the melanoma patients from (j) sequenced before (Pre, n = 11) or during (On, n = 11) kinase inhibition or after resistance (Resist, n = 10) had developed. l Cytolytic activity as the geometric mean of granzyme A and perforin RNA expression in patients from (j). Significance was calculated by t tests in (g), (i), (k) (l) and Kolmogorov–Smirnov-based permutation test as FDR-corrected q-values in (a), (e), (j). All tests are two-sided. Boxplots display median (center line), 25th/75th percentile (lower/upper box hinges), whiskers extend to the most extreme value within 1.5× interquartile range (IQR) of the hinges. Source data are provided as a Source Data file.