Development and implementation of a cell-based assay to discover agonists of the nuclear receptor REV-ERBα

Authors

  • Yuliya Hering Fraunhofer Institute for Molecular Biology and Applied Ecology, ScreeningPort (Fraunhofer-IME SP), Schnackenburgallee 114, D-22525 Hamburg, Germany.
  • Alexandre Berthier University of Lille-EGID, CHU, Institut Pasteur de Lille, INSERM UMR 1011, 1 rue du professeur Calmette, BP245, 59019 Lille, France.
  • Helene Duez University of Lille-EGID, CHU, Institut Pasteur de Lille, INSERM UMR 1011, 1 rue du professeur Calmette, BP245, 59019 Lille, France.
  • Philippe Lefebvre University of Lille-EGID, CHU, Institut Pasteur de Lille, INSERM UMR 1011, 1 rue du professeur Calmette, BP245, 59019 Lille, France.
  • Benoit Deprez University of Lille-EGID, CHU, Institut Pasteur de Lille, INSERM UMR 1011, 1 rue du professeur Calmette, BP245, 59019 Lille, France.
  • Philip Gribbon Fraunhofer Institute for Molecular Biology and Applied Ecology, ScreeningPort (Fraunhofer-IME SP), Schnackenburgallee 114, D-22525 Hamburg, Germany.
  • Markus Wolf Fraunhofer Institute for Molecular Biology and Applied Ecology, ScreeningPort (Fraunhofer-IME SP), Schnackenburgallee 114, D-22525 Hamburg, Germany.
  • Jeanette Reinshagen Fraunhofer Institute for Molecular Biology and Applied Ecology, ScreeningPort (Fraunhofer-IME SP), Schnackenburgallee 114, D-22525 Hamburg, Germany.
  • Francoise Halley Fraunhofer Institute for Molecular Biology and Applied Ecology, ScreeningPort (Fraunhofer-IME SP), Schnackenburgallee 114, D-22525 Hamburg, Germany.
  • Juliane Hannemann Institute of Clinical Pharmacology and Toxicology, University Medical Center Hamburg-Eppendorf, Martinistraße 52, D-20246, Hamburg, Germany.
  • Rainer Böger Institute of Clinical Pharmacology and Toxicology, University Medical Center Hamburg-Eppendorf, Martinistraße 52, D-20246, Hamburg, Germany.
  • Bart Staels University of Lille-EGID, CHU, Institut Pasteur de Lille, INSERM UMR 1011, 1 rue du professeur Calmette, BP245, 59019 Lille, France.
  • Sheraz Gul Fraunhofer Institute for Molecular Biology and Applied Ecology, ScreeningPort (Fraunhofer-IME SP), Schnackenburgallee 114, D-22525 Hamburg, Germany.

DOI:

https://doi.org/10.14440/jbm.2018.244

Keywords:

nuclear receptor, REV-ERBα, luciferase reporter, assay development, drug discovery, high throughput screening

Abstract

The nuclear receptors are transcription factors involved in the regulation of a variety of physiological processes whose activity can be modulated by binding to relevant small molecule ligands. Their dysfunction has been shown to play a role in disease states such as diabetes, cancer, inflammatory diseases, and hormonal resistance ailments, which makes them interesting targets for drug discovery. The nuclear receptor REV-ERBα is involved in regulating the circadian rhythm and metabolism. Its natural ligand is heme and there is significant interest in identifying novel synthetic modulators to serve as tools to characterize its function and to serve as drugs in treating metabolic disorders. To do so, we established a mammalian cell-based two-hybrid assay system capable of measuring the interaction between REV-ERBα and its co-repressor, nuclear co-repressor 1. This assay was validated to industry standard criteria and was used to screen a subset of the LOPAC®1280 library and 29568 compounds from a diverse compound library. Profiling of the primary hits in a panel of counter and selectivity assays confirmed that REV-ERBα activity can be modulated pharmacologically and chemical scaffolds have been identified for optimization.



Author Biography

Yuliya Hering, Fraunhofer Institute for Molecular Biology and Applied Ecology, ScreeningPort (Fraunhofer-IME SP), Schnackenburgallee 114, D-22525 Hamburg, Germany.

Dr. Sheraz Gul is Head of Drug Discovery, Assay Development & Screening at the Fraunhofer Institute for Molecular Biology and Applied Ecology. He is responsible for the global management of drug discovery activities and has 23 years

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Published

2018-07-02

How to Cite

1.
Hering Y, Berthier A, Duez H, Lefebvre P, Deprez B, Gribbon P, Wolf M, Reinshagen J, Halley F, Hannemann J, Böger R, Staels B, Gul S. Development and implementation of a cell-based assay to discover agonists of the nuclear receptor REV-ERBα. J Biol Methods [Internet]. 2018Jul.2 [cited 2021Oct.21];5(3):e94. Available from: https://jbmethods.org/jbm/article/view/244

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