Fig. 6: MTHFD2 inhibitor TH9619 impairs cancer progression in vivo.
a, Cell viability of HL-60 cells on TH7299, TH9028 and TH9619 treatment for 96 h under increasing concentrations of medium folate supplementation. Representatives of three independent experiments are shown. Data are displayed as means (n = 2 independent cell cultures). b, Kaplan–Meier curve showing overall survival of NOG mice with HL-60 IV xenograft tumors after treatment with TH9619 versus vehicle control, on standard chow (SDS) or LF (n (SDS groups) = 6 mice, n (LF groups) = 7 mice). P = 0.001 calculated using a one-way Mantel–Cox log-rank test. c, Plasma concentration of 5-MTHF and thymidine at the time of sacrifice. Data are displayed as median ± s.d. (n = 8 mice per group). **P5-MTHF(SDS) = 0.0015, **P5-MTHF(TH9619 SDS-LF) = 0.0026, **Pthymidine = 0.0013, ****P < 0.0001; two-way ANOVA with Tukey’s correction for multiple comparisons (F5-MTHF(diet) = 63.93, F5-MTHF(treatment) = 11.82, Fthymidine(diet) = 6.307, Fthymidine(treatment) = 46.01, d.f.diet = 1, d.f.treatment = 1); NS, not significant. d, Target engagement of MTHFD2 and TYMS in tumor samples analyzed using CETSA. Representative images are shown for one of two independent experiments. Graphs show the nondenatured target fraction from TH9619-treated animals and vehicle controls from the LF group, displayed as means ± s.d. (n = 4). MTHFD2 melting temperature (ΔTm) = 17 °C, TYMS ΔTm = 1 °C. e, Kaplan–Meier curve showing overall survival of NSG mice on LF with HL-60 IV xenograft tumors after treatment with TH9619 10 mg kg−1 twice daily, TH9619 30 mg kg−1 twice daily, TH9619 60 mg kg−1 four times daily, AraC 50 mg kg−1 five times daily or vehicle control (n = 5 mice per group). There was significantly improved survival in the TH9619 group receiving 60 mg kg−1 four times daily compared with the AraC and vehicle groups. P = 0.0095 calculated using a one-way Mantel–Cox log-rank test.
