Fig. 1

Modelling of drug-induced tumor evolution in vitro, in vivo and “in patients”. a, b Cisplatin treatment of PDPCs that are representative of phenotypically heterogeneous (HN137Pri) or homogeneous (HN120Pri) populations. Drug-induced selection of ECAD+ epithelial cells (green), and the elimination of Vim+ mesenchymal (red) cells, in HN137Pri model (a). De novo emergence of Vim + mesenchymal (red) cells within the phenotypically ECAD + epithelial (green) HN120Pri parental population (b) (n = 24 replicates per experimental condition). c, d Immunofluorescence-based characterization of naive and drug-resistant cells from HN120 and HN137 populations. Cells were stained for the expression of epithelial ECAD (in green) and mesenchymal VIM (in red) markers (c), as well as basal KRT5 (in green) and luminal KRT18 (in red) markers (d). e, f In vivo modelling of cisplatin-resistance in subcutaneous Pdx models of HN120Pri and HN137Pri cells (also see Supplementary Fig. 3a-g). Flow-cytometeric analysis of ECAD and VIMENTIN positive (VIM+) cells reveals a reduction of ECAD + population and the de novo gain of VIM + cells in the HN120 model (e). Loss of VIM + cells and an enrichment of ECAD + population is observed post-cisplatin selection in the HN137 model (f). The gating strategy for viable singlets is exemplified in Supplementary Figure 3a. g “In patient” validation of epithelial (ECAD+) to mesenchymal (VIM+) cell-state switch in HN120Met vs. HN120Pri patient tumor, and the selection of ECAD + epithelial cells with concomitant loss of VIM + mesenchymal cells in cisplatin treated HN137 patient recurrent (and thus resistant) tumor in the clinic. n = 24 replicates (a, b) and n = 3 replicates (c–f) per experimental condition). Scale bar = 100 μM (a, b, g), 50 μM (c–d)