Targeting MYC dependency in ovarian cancer through inhibition of CDK7 and CDK12/13
- Dana-Farber Cancer Institute, United States
- Harvard Medical School, United States
- Baylor College of Medicine, United States
Figures
Figure 1 with 1 supplement
MYC is frequently amplified in ovarian cancer and required for cancer cell growth.
(A) Copy number plots of TCGA high-grade serous ovarian cancer samples for chromosome 8 (top) and part of the q24 arm (bottom). Red color indicates a high chromosomal copy number ratio, blue …
Figure 1—figure supplement 1
Analysis of MYC dependency in cancer cell lines.
Data were downloaded from https://depmap.org/portal/, and analyzed in GraphPad Prism 7. (A) Lack of correlation between MYC gene copy number and MYC dependency scores. (B) MYC dependency is highly …
Figure 2 with 2 supplements
Targeted screen of transcriptional and epigenetic regulators identifies THZ1 as key transcriptional regulator of MYC.
(A) Schematic diagram of a screen for small molecules that inhibit MYC expression. Ovarian cancer cells were treated with selected epigenetic/transcriptional inhibitors for 6 hr at a final …
Figure 2—figure supplement 1
Small molecules screen for compounds inhibiting MYC expression.
(A-B) Ovarian cancer cell lines, KURAMOCHI (A) or COV362 (B), were treated with indicated inhibitors for 6 hr, followed by preparation of whole cell lysates and immunoblotting using the indicated …
Figure 2—figure supplement 2
THZ1 inhibits the expression of both MYC and MCL1.
(A) Kaplan-Meier plot of disease-free survival in ovarian cancer patients with (red) or without (blue) co-amplification of MYC and MCL-1 (left panel). Kaplan-Meier plot of disease-free survival in …
Figure 3 with 2 supplements
THZ1 represses MYC target genes in ovarian cancer cells.
(A) Heatmaps showing row normalized gene expression for all actively expressed genes (n = 14,068) (cell count normalized FPKM > 1 in at least one sample). (B) Scatter plots comparing change in gene …
Figure 3—figure supplement 1
PVT1, a lncRNA gene co-amplified with MYC on 8q24, is not significantly downregulated by THZ1.
Scatter plots comparing change in gene expression upon THZ1 treatment to gene expression levels in DMSO, in KUROMOCHI (left) and COV362 cells (right). X-axis shows the Log2 fold change of expression …
Figure 3—figure supplement 2
THZ1 selectively downregulates MYC target genes.
Bar plot showing gene sets from the MSigDB Hallmarks of Cancer that are significantly enriched as differentially downregulated upon 250 nM THZ1 treatment in Kuramochi and Cov362 cells (FDR q value …
Figure 4 with 2 supplements
Downregulation of MYC and MCL-1 requires co-inhibition of CDK7 and CDK12/13 in KURAMOCHI cells.
(A) Chemical structures of inhibitors and their primary protein targets. (B) KURAMOCHI cells were treated with increasing concentrations of YKL-1–116 or THZ531, with cell lysates prepared following …
Figure 4—figure supplement 1
Downregulation of MYC and MCL-1 requires co-inhibition of CDK7 and CDK12/13 in COV362 cells.
(A) COV362 cells were treated with increasing concentrations of YKL-1–116 or THZ531, with cell lysates prepared following 6 hr of treatment. (B) qPCR analysis of MYC in ovarian cancer cells treated …
Figure 4—figure supplement 2
The mutant CDK7 (C312S) rescued THZ1-induced MYC downregulation and cell growth inhibition by YKL-1–116.
OVCAR8 cells were infected with or without HA-tagged wild-type CDK7 (WT) or C312S mutant (CS), followed by G418 selection to obtain stably transduced cells. The cells were pre-treated with …
Figure 5 with 1 supplement
THZ1 abrogates the growth of patient-derived high-grade serous ovarian cancer cells in vivo.
(A) A schematic diagram of the experimental design. (B) Luminescence picture of luciferized PDX mice (eg. DF-106) at day 0 and day 27, treated with vehicle control (10 ml/kg, PO, QD) or THZ1 (10 …
Figure 5—figure supplement 1
Body weight measurement of the 11 PDX models (n = 44 mice) treated with vehicle control (10 ml/kg, PO, QD) or THZ1 (10 mg/kg, IP, BID) or Olaparib (100 mg/kg, PO, QD) or Combo.
https://doi.org/10.7554/eLife.39030.014
Author response image 1
CDK7 inhibitor (YKL-1-116) and CDK12/13 inhibitor (THZ531) have different effects on MYC expression.
Indicated ovarian cancer cell lines were treated with increasing concentrations of YKL- 1-116 or THZ531, with cell lysates prepared following 6 hours of treatment.
Author response image 2
Overexpressing CDK7 C312S mutant rescued cell growth inhibition by YKL-1-116.
Author response image 3
Overexpression of GFP/ MYC/ MYC-1/ MYC+MCL-1 in OVCAR8 cells.
Author response image 4
Overexpression of MYC inhibits cancer cell growth.
OVCAR8 cells were infected with pCDH-GFP/ MYC/ MYC-1/ MYC+MCL-1 twice, then selected with puromycin 1 ug/ml for 2 days. Cells were seeded at 5,000 per well in a 12-well plate, then treated with DMSO …
Additional files
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Supplementary file 1
Table of epigenetic and transcriptional compound screen.
- https://doi.org/10.7554/eLife.39030.016
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Supplementary file 2
Table of normalized MYC expression values.
- https://doi.org/10.7554/eLife.39030.017
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Transparent reporting form
- https://doi.org/10.7554/eLife.39030.018
