Fig. 4: Characterization and therapeutic intervention of residual disease in human NSCLC PDO and xenograft mouse model systems.

a Schematic presentation for the generation of persister cells in NSCLC patient-derived organoid (PDO) models. Schematic diagram was created with BioRender.com. b YAP cluster 3 candidate genes were selected YAP genes based on the time-dependent modulation after 2 uM osimertinib treatment (please see Supplementary Fig. 5e, f). Mean expression of cluster 3 genes in 0.1% DMSO control (DMSO) and drug-tolerant persisters (DTP) across treatment-sensitive EGFR-mutant PDO models, i.e., EGFR^del19 TH107 and EGFR^L858R TH330. Statistical analysis by n = 3 independent experiments, mean ± s.d., Statistical significance is indicated by two-way ANOVA test. The box plot displays 25th (lower bound), 50th (centre, median), and 75th (upper bound) percentiles, with whiskers (minima (bottom), maxima (top)) extending 1.5 * IQR. c Treatment response to escalating doses of osimertinib upon combinatorial treatment with FAK inhibitor VS-4718 (1 μM) in EGFR^del19 TH107 DTPs as determined by CellTiter-Glo assay. Statistical analysis by n = 3 independent experiments. d, e Immunohistochemistry staining for YAP in residual tumors cells of EGFR-mutant TH021 and ALK fusion-positive LG0812 PDX models upon treatment with targeted inhibitors (TKI) compared to vehicle control (VEH). TH021 and LG0812 images are representative of total n = 3 and n = 4 independent experiments, respectively. Quantification of nuclear levels (% nuclear) by automated image analysis. Statistical evaluation by two-sided t-test. For TH021: VEH vs TKI, * p = 0.0127. For LG0812: VEH vs TKI, ** p = 0.0022. Arrows indicate YAP-positive tumor cell nuclei. f Relative tumor volume changes in an EGFR-mutant H1975 xenograft (CDX) mouse model across vehicle, 5 mg/kg osimertinib, 50 mg/kg FAK inhibitor VS-4718, and 5 mg/kg osimertinib + 50 mg/kg FAK inhibitor VS-4718 treatment groups. Statistical analysis by n = 10 mice. Statistical evaluation by Mann–Whitney U test, **p = ≤0.0021 (two-tailed). The box plot displays the minimum (lower bound), median (centre), and maximum (upper bound). g–j The combination therapies with 2.5 mg/kg osimertinib and 30 mg/kg TEAD inhibitor VT108 induce more durable response and impair tumor regrowth in (g) H1975 CDX model (n = 8 mice), (h) TH021 PDX model (n = 8 mice), (i) LU-01-1291 PDX model(n = 8 mice) and (j) LU1868 PDX model (n = 10 mice). For g H1975 CDX model, all treatments were continued throughout the entire study. For h–j PDX models, the gray area indicates the treatment duration, then discontinuation on day 21 for TH021, day 30 for LU-01-1291, and day 38 for LU1868 PDX models.