TY - JOUR
T1 - Predicting novel therapies and targets
T2 - Regulation of Notch3 by the bromodomain protein Brd4
AU - Villar-Prados, Alejandro
AU - Wu, Sherry Y.
AU - Court, Karem A.
AU - Ma, Shaolin
AU - LaFargue, Christopher
AU - Chowdhury, Mamur A.
AU - Engelhardt, Margaret I.
AU - Ivan, Cristina
AU - Ram, Prahlad T.
AU - Wang, Ying
AU - Baggerly, Keith
AU - Rodriguez-Aguayo, Cristian
AU - Lopez-Berestein, Gabriel
AU - Yang, Shyh Ming
AU - Maloney, David J.
AU - Yoshioka, Makoto
AU - Strovel, Jeffrey W.
AU - Roszik, Jason
AU - Sood, Anil K.
N1 - Funding Information:
Portions of the research reported in this publication were supported by the NIH (P30 CA016672, UH3TR000943, P50 CA217685, U01 CA213759 P50 CA098258, and R35 CA209904) and the Partnership for Excellence in Cancer Research (U54CA096300/U54CA096297). Additional portions of this work were also funded by the Ovarian Cancer Moon Shot, the Blanton-Davis Ovarian Cancer Research Program, American Cancer Society Research Professor Award, and the Frank McGraw Memorial Chair in Cancer Research. We acknowledge members of The University of Texas MD Anderson Cancer Center cores including: the Flow Cytometry and Cellular Imaging Core Facility, the Characterized Cell Line Core Facility; the Research Histology, Pathology, and Imaging Core; the Small Animal Imaging Facility core; and the RPPA Core Facility. These shared resources are partially funded by the NCI Cancer Support Grant, P30CA16672. We also want to acknowledge the University of Texas MD Anderson Cancer Center Department of Scientific Publications for aiding in editing this manuscript. We acknowledge ConverGene for the development and use of CN210. SYW was supported by the CPRIT Research Training Program (RP101502, RP140106, and RP170067).
Publisher Copyright:
© 2018 American Association for Cancer Research.
PY - 2019/2
Y1 - 2019/2
N2 - Systematic approaches for accurate repurposing of targeted therapies are needed. We developed and aimed to biologically validate our therapy predicting tool (TPT) for the repurposing of targeted therapies for specific tumor types by testing the role of Bromodomain and Extra-Terminal motif inhibitors (BETi) in inhibiting BRD4 function and downregulating Notch3 signaling in ovarian cancer. Utilizing established ovarian cancer preclinical models, we carried out in vitro and in vivo studies with clinically relevant BETis to determine their therapeutic effect and impact on Notch3 signaling. Treatment with BETis or siRNA-mediated BRD4 knockdown resulted in decreased cell viability, reduced cell proliferation, and increased cell apoptosis in vitro. In vivo studies with orthotopic mouse models demonstrated that treatment with BETi decreased tumor growth. In addition, 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. Our findings provide biological validation for the TPT by demonstrating that BETis can be an effective therapeutic agent for ovarian cancer by downregulating Notch3 expression. The TPT could rapidly identify candidate drugs for ovarian or other cancers along with novel companion biomarkers.
AB - Systematic approaches for accurate repurposing of targeted therapies are needed. We developed and aimed to biologically validate our therapy predicting tool (TPT) for the repurposing of targeted therapies for specific tumor types by testing the role of Bromodomain and Extra-Terminal motif inhibitors (BETi) in inhibiting BRD4 function and downregulating Notch3 signaling in ovarian cancer. Utilizing established ovarian cancer preclinical models, we carried out in vitro and in vivo studies with clinically relevant BETis to determine their therapeutic effect and impact on Notch3 signaling. Treatment with BETis or siRNA-mediated BRD4 knockdown resulted in decreased cell viability, reduced cell proliferation, and increased cell apoptosis in vitro. In vivo studies with orthotopic mouse models demonstrated that treatment with BETi decreased tumor growth. In addition, 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. Our findings provide biological validation for the TPT by demonstrating that BETis can be an effective therapeutic agent for ovarian cancer by downregulating Notch3 expression. The TPT could rapidly identify candidate drugs for ovarian or other cancers along with novel companion biomarkers.
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U2 - 10.1158/1535-7163.MCT-18-0365
DO - 10.1158/1535-7163.MCT-18-0365
M3 - Article
C2 - 30420565
AN - SCOPUS:85061052323
SN - 1535-7163
VL - 18
SP - 421
EP - 436
JO - Molecular cancer therapeutics
JF - Molecular cancer therapeutics
IS - 2
ER -