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Published on: July 25, 2020
Preclinical Evaluation of NTX-301, a Novel DNA Hypomethylating Agent in Ovarian Cancer
Yinu Wang1, Xiaolei Situ1, Horacio Cardenas1
1Department of Obstetrics and Gynecology, Feinberg School of Medicine, Northwestern University, Chicago, Illinois.
Purpose:
DNA methylation causes silencing of tumor-suppressor and differentiation-associated genes, being linked to chemoresistance. Previous studies demonstrated that hypomethylating agents (HMA) resensitize ovarian cancer to chemotherapy. NTX-301 is a highly potent and orally bioavailable HMA, in early clinical development.
Experimental Design:
The antitumor effects of NTX-301 were studied in ovarian cancer models by using cell viability, stemness and ferroptosis assays, RNA sequencing, lipidomic analyses, and stimulated Raman spectroscopy.
Results:
Ovarian cancer cells (SKOV3, IC50 = 5.08 nmol/L; OVCAR5 IC50 = 3.66 nmol/L) were highly sensitive to NTX-301 compared with fallopian tube epithelial cells. NTX-301 downregulated expression of DNA methyltransferases 1-3 and induced transcriptomic reprogramming with 15,000 differentially expressed genes (DEG, P < 0.05). Among them, Gene Ontology enrichment analysis identified regulation of fatty acid biosynthesis and molecular functions related to aldehyde dehydrogenase (ALDH) and oxidoreductase, known features of cancer stem cells. Low-dose NTX-301 reduced the ALDH(+) cell population and expression of stemness-associated transcription factors. Stearoyl-coenzyme A desaturase 1 (SCD), which regulates production of unsaturated fatty acids (UFA), was among the top DEG downregulated by NTX-301. NTX-301 treatment decreased levels of UFA and increased oxidized lipids, and this was blunted by deferoxamine, indicating cell death via ferroptosis. NTX-301-induced ferroptosis was rescued by oleic acid. In vivo, monotherapy with NTX-301 significantly inhibited ovarian cancer and patient-derived xenograft growth (P < 0.05). Decreased SCD levels and increased oxidized lipids were detected in NTX-301-treated xenografts.
Conclusions:
NTX-301 is active in ovarian cancer models. Our findings point to a new mechanism by which epigenetic blockade disrupts lipid homeostasis and promotes cancer cell death.
Insights
NTX-301, a potent hypomethylating agent, shows significant antitumor activity in ovarian cancer models by disrupting lipid homeostasis and inducing ferroptosis. This epigenetic therapy offers a novel approach to combatting chemoresistant ovarian cancers.
Area of Science:
- Epigenetics
- Cancer Biology
- Lipid Metabolism
Background:
- DNA methylation is linked to gene silencing and chemoresistance in ovarian cancer.
- Hypomethylating agents (HMA) can resensitize ovarian cancer to chemotherapy.
Purpose of the Study:
- To evaluate the antitumor effects of NTX-301, a novel HMA, in ovarian cancer models.
- To elucidate the mechanism of action of NTX-301, including its impact on cancer stemness and cell death pathways.
Main Methods:
- Cell viability, stemness, and ferroptosis assays were performed.
- RNA sequencing, lipidomic analyses, and stimulated Raman spectroscopy were utilized.
- In vivo studies included ovarian cancer and patient-derived xenograft models.
Main Results:
- NTX-301 demonstrated high sensitivity in ovarian cancer cells, downregulating DNA methyltransferases and inducing significant transcriptomic reprogramming.
- NTX-301 reduced cancer stem cell populations and downregulated stearoyl-coenzyme A desaturase 1 (SCD).
- NTX-301 treatment led to decreased unsaturated fatty acids, increased oxidized lipids, and induced ferroptosis, which was rescued by oleic acid. Significant inhibition of tumor growth was observed in vivo.
Conclusions:
- NTX-301 exhibits potent activity in ovarian cancer models.
- Epigenetic blockade by NTX-301 disrupts lipid homeostasis, promoting cancer cell death via ferroptosis.
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