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A Novel Flow Cytometric Assay to Identify Inhibitors of RBPJ-DNA Interactions.
SLAS Discov. 2020 Jun 22 [Online ahead of print]SD

Abstract

Notch signaling is often involved in cancer cell initiation and proliferation. Aberrant Notch activation underlies more than 50% of T-cell acute lymphoblastic leukemia (T-ALL); accordingly, chemicals disrupting Notch signaling are of potential to treat Notch-dependent cancer. Here, we developed a flow cytometry-based high-throughput assay to identify compounds that disrupt the interactions of DNA and RBPJ, the major downstream effector of Notch signaling. From 1492 compounds, we identified 18 compounds that disrupt RBPJ-DNA interactions in a dose-dependent manner. Cell-based assays further revealed that auranofin downregulates Notch-dependent transcription and decreases RBPJ-chromatin interactions in cells. Most strikingly, T-ALL cells that depend on Notch signaling for proliferation are more sensitive to auranofin treatment, supporting the notion that auranofin downregulates Notch signaling by disrupting RBPJ-DNA interaction. These results validate the feasibility of our assay scheme to screen for additional Notch inhibitors and provide a rationale to further test the use of auranofin in treating Notch-dependent cancer.

Authors+Show Affiliations

Department of Internal Medicine, Division of Molecular Medicine, Program in Cellular and Molecular Oncology, University of New Mexico Comprehensive Cancer Center, Albuquerque, NM, USA.Department of Pathology, Program in Cancer Therapeutics, University of New Mexico Comprehensive Cancer Center, Albuquerque, NM, USA. Center for Molecular Discovery, University of New Mexico Health Science Center, Albuquerque, NM, USA.Department of Internal Medicine, Division of Molecular Medicine, Program in Cellular and Molecular Oncology, University of New Mexico Comprehensive Cancer Center, Albuquerque, NM, USA.Department of Internal Medicine, Division of Molecular Medicine, Program in Cellular and Molecular Oncology, University of New Mexico Comprehensive Cancer Center, Albuquerque, NM, USA.Department of Pathology, Program in Cancer Therapeutics, University of New Mexico Comprehensive Cancer Center, Albuquerque, NM, USA. Center for Molecular Discovery, University of New Mexico Health Science Center, Albuquerque, NM, USA.Department of Internal Medicine, Division of Molecular Medicine, Program in Cellular and Molecular Oncology, University of New Mexico Comprehensive Cancer Center, Albuquerque, NM, USA.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

32567455

Citation

Lake, Robert J., et al. "A Novel Flow Cytometric Assay to Identify Inhibitors of RBPJ-DNA Interactions." SLAS Discovery : Advancing Life Sciences R & D, 2020, p. 2472555220932552.
Lake RJ, Haynes MK, Dreval K, et al. A Novel Flow Cytometric Assay to Identify Inhibitors of RBPJ-DNA Interactions. SLAS Discov. 2020.
Lake, R. J., Haynes, M. K., Dreval, K., Bilkis, R., Sklar, L. A., & Fan, H. Y. (2020). A Novel Flow Cytometric Assay to Identify Inhibitors of RBPJ-DNA Interactions. SLAS Discovery : Advancing Life Sciences R & D, 2472555220932552. https://doi.org/10.1177/2472555220932552
Lake RJ, et al. A Novel Flow Cytometric Assay to Identify Inhibitors of RBPJ-DNA Interactions. SLAS Discov. 2020 Jun 22;2472555220932552. PubMed PMID: 32567455.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A Novel Flow Cytometric Assay to Identify Inhibitors of RBPJ-DNA Interactions. AU - Lake,Robert J, AU - Haynes,Mark K, AU - Dreval,Kostiantyn, AU - Bilkis,Rabeya, AU - Sklar,Larry A, AU - Fan,Hua-Ying, Y1 - 2020/06/22/ PY - 2020/6/23/entrez KW - Notch inhibitor KW - RBPJ-DNA interactions KW - T-ALL KW - auranofin KW - flow cytometry-based high-throughput screening SP - 2472555220932552 EP - 2472555220932552 JF - SLAS discovery : advancing life sciences R & D JO - SLAS Discov N2 - Notch signaling is often involved in cancer cell initiation and proliferation. Aberrant Notch activation underlies more than 50% of T-cell acute lymphoblastic leukemia (T-ALL); accordingly, chemicals disrupting Notch signaling are of potential to treat Notch-dependent cancer. Here, we developed a flow cytometry-based high-throughput assay to identify compounds that disrupt the interactions of DNA and RBPJ, the major downstream effector of Notch signaling. From 1492 compounds, we identified 18 compounds that disrupt RBPJ-DNA interactions in a dose-dependent manner. Cell-based assays further revealed that auranofin downregulates Notch-dependent transcription and decreases RBPJ-chromatin interactions in cells. Most strikingly, T-ALL cells that depend on Notch signaling for proliferation are more sensitive to auranofin treatment, supporting the notion that auranofin downregulates Notch signaling by disrupting RBPJ-DNA interaction. These results validate the feasibility of our assay scheme to screen for additional Notch inhibitors and provide a rationale to further test the use of auranofin in treating Notch-dependent cancer. SN - 2472-5560 UR - https://www.unboundmedicine.com/medline/citation/32567455/A_Novel_Flow_Cytometric_Assay_to_Identify_Inhibitors_of_RBPJ-DNA_Interactions L2 - https://journals.sagepub.com/doi/10.1177/2472555220932552?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -
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