Predicting novel therapies and targets: Regulation of Notch3 by the bromodomain protein BRD4.

Therapeutic Approaches

Abstract

BACKGROUND: Systematic approaches for accurate re-purposing of targeted therapies are needed. We developed and aimed to biologically validate our Therapy Predicting Tool (TPT) for the re-purposing of targeted therapies for specific tumor types by testing the role of Bromodomain and Extra-Terminal motif inhibitors (BETis) in inhibiting BRD4 function and downregulating Notch3 signaling in ovarian cancer. METHODS: Utilizing established ovarian cancer pre-clinical models, we carried out in vitro and in vivo studies using clinically relevant BETis to determine their therapeutic effect and impact on Notch3 signaling. RESULTS: Treatment with BETis or siRNA mediated BRD4 knockdown resulted in decreased cell viability, reduced cell proliferation and increased cell apoptosis in vitro. In vivo with orthotopic mouse models demonstrated that treatment with BETi decreased tumor growth. Additionally, knockdown of BRD4 with doxycycline inducible shRNA increased survival up to 50% (p < 0.001). Treatment with either BETis or BRD4 siRNA decreased Notch3 expression both in vitro and in vivo. BRD4 inhibition also decreased the expression of NOTCH3 targets, including HES1. Chromatin immunoprecipitation revealed that BRD4 was present at the NOTCH3 promoter. CONCLUSION: Our findings provide biological validation for the TPT by demonstrating that BETis can be an effective therapeutic for ovarian cancer by downregulating Notch3 expression. IMPACT: The TPT could rapidly identify candidate drugs for ovarian or other cancers along with novel companion biomarkers.

Authors

Villar-Prados, Alejandro; Wu, Sherry Y; Court, Karem A; Ma, Shaolin; LaFargue, Christopher; Chowdhury, Mamur A; Engelhardt, Margaret I; Ivan, Cristina; Ram, Prahlad T; Wang, Ying; Baggerly, Keith; Rodriguez-Aguayo, Cristian; Lopez-Berestein, Gabriel; Yang, Shyh Ming; Maloney, David J; Yoshioka, Makoto; Strovel, Jeffrey W; Roszik, Jason; Sood, Anil K;

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