Fig. 6: TC behavior in the metastatic niche is predetermined by methylation state.
From: Lung endothelium exploits susceptible tumor cell states to instruct metastatic latency
a, Gene scores of EMT (left)- and Wnt pathway-associated genes (right) in 362 cultured TCs visualized in UMAP. b, Correlation of gene scores. P and r values by Pearson correlation. c, Violin plot of EMT gene scores in cultured TCs with Wnt gene score >0 (Wnt high, cyan) and Wnt gene score <0 (Wnt low, black). d, Left, schematic of experiment. 4T1-GFP cells were either treated overnight (O/N) with Wnt agonist (pulse) or for 2 weeks (reprogrammed) before injection. Relative fraction of extravasated TCs normalized to the respective control 1.5 days postinjection (middle) and relative fraction of LTCs normalized to the respective control 3.5 days postinjection (right). Data presented as mean ± s.d., P values by two-way ANOVA with Sidak post test; n = 4–6 mice. e, Left, schematic of experiment. 4T1-GFP cells were treated overnight with the demethylating agent decitabine. Percentage of extravasated TCs for control and hypomethylation (hypo) treatment 1.5 days postinjection (middle) and percentage of latent TCs 3.5 days postinjection (right). Data presented as mean ± s.d., P values by two-tailed t-test; n = 6 mice. f, Left, schematic of experimental L-O-F approach. 4T1-GFP cells were treated overnight with decitabine. Gray bar indicates time span (in days) of daily treatment with LGK974. Percentage of latent TCs 3.5 days postinjection for control and LGK974-treated animals (middle) and relative TC number normalized to EC abundance in lungs (right). Data presented as mean ± s.d., P values by two-tailed t-test; n = 5–6 mice. g, Scatter plot of methylation level (fraction of methylated CpG islands) for gene bodies in latent and proliferative TCs. Solid red line indicates no differences in methylation, dotted red lines indicate thresholds for >10% differences in methylation.
