Fig 1.
ONC201 modulates stem cell-related gene expression.
(A) Summary of targeted network analysis of stem cell-related changes in ONC201-treated (10 μM) HCT116 p53-null cells by Ingenuity Pathway Analysis. The—log(p-value) is indicated for each group of genes. Ratio indicates the relative number of genes that were significantly changed upon ONC201 treatment compared to total number of genes in the group. (B) qRT-PCR for indicated stem cell-related genes in DMSO/ONC201-treated (5 μM, 18 h/48 h, n = 3) HCT116 p53-null cells. * indicates p < 0.02 relative to DMSO.
Table 1.
ONC201-mediated CSC- and Wnt-pathway-related changes in gene expression.
Table 2.
ONC201-mediated stem cell-related changes in gene expression.
Fig 2.
ONC201 targets CSCs in prostate and glioblastoma tumors.
(A) Effect of indicated concentrations of ONC201 (72 h) on viability of newly diagnosed (GBM8, GBM18) and recurrent (GBM67R, GBM 152) glioblastoma cells in 3D neurosphere culture. (B) qRT-PCR for indicated stem cell-related genes in DMSO/ONC201-treated (5 μM, 24 h/48 h, n = 3) SNB19 cells. * indicates p < 0.0002 relative to DMSO. (C) Effect of DMSO/ONC201 (5 μM, 72 h, n = 3) on tumor sphere formation of indicated prostate cancer cell lines. * indicates p < 0.025 relative to DMSO. (D) qRT-PCR for indicated stem cell-related genes in DMSO/ONC201-treated (5 μM, 24 h/48 h, n = 3) DU145 cells. * indicates p < 0.04 relative to DMSO.
Fig 3.
Inhibition of CSCs does not occur in tumor cells with acquired resistance to ONC201.
(A) Effect of DMSO/ONC201 (5 μM, 72 h, n = 3) on tumor sphere formation of RKO wild-type (wt) and ONC201-resistant (resist) cells. Representative image (10X magnification) of spheres (> 60 μm) (B) Quantification of spheres in (A). (C) qRT-PCR for indicated stem cell-related genes in DMSO/ONC201-treated (5 μM, 48 h, n = 3) RKO wild-type (wt) and ONC201-resistant (resist) cells. # indicates p < 0.003 relative to wt DMSO. * indicates p < 0.05 relative to wt ONC201. (D) Western blot for indicated stem cell-related proteins in DMSO/ONC201-treated (5 μM, 72 h) RKO wild-type (wt) and ONC201-resistant (resist) cells.
Fig 4.
CSC-related gene expression in solid tumors as a potential biomarker of response for ONC201.
Distribution of ONC201 efficacy (IC50) in >1000 GDSC cell lines based on basal RNA expression of (A) TCF3, (B) HES7. (C) Distribution of ONC201 efficacy (IC50) in >1000 GDSC cell lines based on fulfillment of at least two expression based criteria (low expression of ID1/CD44 and high expression of TCF3/HES7) against cell lines that fulfilled none. P value and D statistic are indicated.