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Methods for measuring HMGB1 release during immunogenic cell death.

Liwei Zhao1, Peng Liu1, Oliver Kepp1

  • 1Faculty of Medicine, University of Paris Sud, Kremlin-Bicêtre, France; Cell Biology and Metabolomics Platforms, Gustave Roussy Cancer Campus, Villejuif, France; Equipe 11 labellisée Ligue Nationale contre le Cancer, Centre de Recherche des Cordeliers, Paris, France; Equipe labellisée Ligue Nationale Contre le Cancer, Université Paris Descartes, Université Sorbonne Paris Cité, Université Paris Diderot, Institut National de la Santé et de la Recherche Médicale (INSERM), UMR1138, Centre de Recherche des Cordeliers, Paris, France; Université Paris Descartes, Sorbonne Paris Cité, Paris, France; Université Pierre et Marie Curie, Paris, France.

Methods in Enzymology
|November 16, 2019
PubMed
Summary
This summary is machine-generated.

Researchers developed a new screening method to find drugs that release high mobility group box 1 (HMGB1), a key danger signal. This method uses a novel "retention using selective hooks" (RUSH) system for efficient drug discovery.

Keywords:
AnthracyclinesCancerHigh-throughput screeningImmunogenic cell death (ICD)Retention using selective hooks (RUSH) assay

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Area of Science:

  • Cell Biology
  • Immunology
  • Pharmacology

Background:

  • High mobility group box 1 (HMGB1) is an alarmin released from cells as a danger signal, activating innate immunity.
  • Extracellular HMGB1 interacts with pattern recognition receptors, stimulating immune responses.

Purpose of the Study:

  • To develop a discovery pipeline for identifying pharmacological agents that induce HMGB1 release.
  • To establish a high-throughput screening method for HMGB1-releasing compounds.

Main Methods:

  • Utilized the "retention using selective hooks" (RUSH) system with a streptavidin-NLS3 fusion protein as a nuclear hook.
  • Synchronized HMGB1 increase by sequestering HMGB1-SBP-GFP, releasing it upon biotin addition.
  • Employed immunogenic cell death (ICD) inducers like anthracyclines to trigger nucleo-cytoplasmic translocation of HMGB1-SBP-GFP.

Main Results:

  • The RUSH system enabled synchronized HMGB1 release upon stimulation.
  • Successfully demonstrated the system's capability to identify HMGB1-releasing agents.
  • Facilitated medium- to high-throughput screening for novel HMGB1-releasing compounds.

Conclusions:

  • The developed RUSH system provides an efficient platform for discovering drugs that modulate HMGB1 release.
  • This pipeline aids in identifying compounds with potential immunomodulatory properties through HMGB1 release.