Extended Data Fig. 10: Vortioxetine reduces tumor burden in vivo independent of serotonin modulation and affects tumor invasiveness and long term growth.
a, Representative MRI images of three ZH-161 transplanted mice (columns) after 15 days of drug treatment (Trial II; n = 7 drugs). Tumor perimeters indicated in yellow. b, Quantification of tumor perimeters corresponding to a. Dots: individual mice per drug (columns); Red lines: mean values. Two-tailed t-test. c, Spheroid formation analyzed by the 2D-projected area of the ZH-562 line measured after 12 days of Vortioxetine treatment (0.1-5 µM; n = 45-47 wells/condition). Data is shown as a boxplot, individual data points, and histogram. d, Number of migrated cells in a collagen-based spheroid invasion assay after 36 h of Vortioxetine treatment (2, 3.5, 5 µM) across four glioblastoma cell lines; LN-229 (n = 560-1125 cells/well), LN-308 (n = 137-426 cells/well), ZH-161 (n = 200-574 cells/well), ZH-562 (n = 38-253 cells/well). e, Mean cell migration distance per condition (n = 5 replicate wells) for d. c-e, One-tailed t-test with adjusted P-values after Holm correction. f, Clonogenic survival measured by a resazurin-based cell viability assay after 11-13 days of Vortioxetine treatment (7 concentrations; 0.625-20 µM, n = 6 replicate wells/concentration) across four glioblastoma cell lines; LN-229 (n = 50 cells/well), LN-308 (n = 300 cells/well), ZH-161 (n = 500 cells/well), and ZH-562 (n = 500 cells/well). Dose-response fitted with a two-parameter log-logistic distribution with 95% confidence intervals (grey) and ED50 (dashed lines). g, Representative immunohistochemistry images of brain sections (n = 3 mice/treatment group) stained with human-specific Ki67 and Vimentin (VIM). h, Ki67 tumor intensity normalized to background with n = 3-4 mice (dots) analyzed per group. Two-tailed t-test comparing CITA and VORT treatment to (-) ctrl. i, Vortioxetine ex vivo PCY score (n = 27 patients; prospective cohort) stratified by Ki67 levels and EGFR CNV alterations. Group 2 patients with low Ki67 levels and an absence of EGFR CNV alterations (n = 7/27; 26%) were significantly less likely to respond to Vortioxetine ex vivo compared to Group 1 (Wilcoxon test; P = 0.011). Among the clinical/genetic parameters in Fig. 2d, e, Ki67 and EGFR alterations were the most predictive two parameters based on a regression subset selection for ex vivo Vortioxetine response. c, d, e, i, h, Boxplots as in Fig. 1b.
