Identification of agents that promote endoplasmic reticulum stress using an assay that monitors luciferase secretion

J Biomol Screen. 2014 Apr;19(4):575-84. doi: 10.1177/1087057113517549. Epub 2013 Dec 26.

Abstract

Disruption of protein processing in the secretory pathway is a measurable hallmark of endoplasmic reticulum (ER) stress. Activation of ER stress-mediated pathways has been implicated in numerous diseases, including cancer. To identify agents that induce ER stress, we established a screen for compounds that reduce secretion of the reporter protein Gaussia luciferase (GLUC). Given the clinically validated importance of targeting ER stress-mediated pathways in the treatment of multiple myeloma (MM), we used this hematological malignancy as a model for validating our screening system. From a screen of 2000 marketed drugs and natural compounds in KMS11 and ARP1 MM cells, we identified 97 agents that reduced GLUC secretion in both cell lines by at least 30%. To confirm inducers of ER stress, we applied a secondary screen that assessed splicing of the unfolded protein response (UPR) transcription factor XBP1. One agent, theaflavin-3,3'-digallate (TF-3), was chosen based on its history of safe human consumption and further validated through studies of ER stress-related pathways, including the UPR and apoptosis. Given these promising results, this screen could be a useful tool to identify agents targeting ER stress-related mechanisms in other cellular systems wherein ER stress plays a role in disease etiology.

Keywords: Gaussia luciferase; endoplasmic reticulum stress; multiple myeloma; protein secretion.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Apoptosis / genetics
  • Cell Line
  • Drug Discovery*
  • Drug Evaluation, Preclinical / methods*
  • Endoplasmic Reticulum Stress / drug effects*
  • Gene Expression Regulation / drug effects*
  • Genes, Reporter*
  • Humans
  • Luciferases / genetics*
  • Luciferases / metabolism
  • Multiple Myeloma / metabolism
  • Secretory Pathway / drug effects
  • Small Molecule Libraries

Substances

  • Small Molecule Libraries
  • Luciferases