title: Identification of agents that promote endoplasmic reticulum stress using an assay that monitors luciferase secretion.
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.
authors.affiliation: 1The Ronald O. Perelman Department of Dermatology, New York University School of Medicine, New York, NY, USA.
authors.forename: Nicole A
authors.initials: NA
authors.lastname: Doudican
authors.forename: Shih Ya
authors.initials: SY
authors.lastname: Wen
authors.forename: Amitabha
authors.initials: A
authors.lastname: Mazumder
authors.forename: Seth J
authors.initials: SJ
authors.lastname: Orlow
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000235:genetics; D002460:Cell Line; D055808:Drug Discovery; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D059865:Endoplasmic Reticulum Stress / Q000187:drug effects*; D005786:Gene Expression Regulation / Q000187:drug effects*; D017930:Genes, Reporter; D006801:Humans; D008156:Luciferases / Q000235:genetics* / Q000378:metabolism; D009101:Multiple Myeloma / Q000378:metabolism; D055571:Secretory Pathway / Q000187:drug effects; D054852:Small Molecule Libraries
chemical_list: D054852:Small Molecule Libraries; D008156:Luciferases
keywords: Gaussia luciferase; endoplasmic reticulum stress; multiple myeloma; protein secretion