title: A High-Throughput Screen Identifies 2,9-Diazaspiro[5.5]Undecanes as Inducers of the Endoplasmic Reticulum Stress Response with Cytotoxic Activity in 3D Glioma Cell Models.
abstract: The endoplasmic reticulum (ER) is involved in Ca2+ signaling and protein folding. ER Ca2+ depletion and accumulation of unfolded proteins activate the molecular chaperone GRP78 (glucose-regulated protein 78) which in turn triggers the ER stress response (ERSR) pathway aimed to restore ER homeostasis. Failure to adapt to stress, however, results in apoptosis. We and others have shown that malignant cells are more susceptible to ERSR-induced apoptosis than their normal counterparts, implicating the ERSR as a potential target for cancer therapeutics. Predicated on these findings, we developed an assay that uses a GRP78 biosensor to identify small molecule activators of ERSR in glioma cells. We performed a quantitative high-throughput screen (qHTS) against a collection of ~425,000 compounds and a comprehensive panel of orthogonal secondary assays was formulated for stringent compound validation. We identified novel activators of ERSR, including a compound with a 2,9-diazaspiro[5.5]undecane core, which depletes intracellular Ca2+ stores and induces apoptosis-mediated cell death in several cancer cell lines, including patient-derived and 3D cultures of glioma cells. This study demonstrates that our screening platform enables the identification and profiling of ERSR inducers with cytotoxic activity and advocates for characterization of these compound in in vivo models.
authors: National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, 20850, United States of America.
authors: Natalia J
authors: NJ
authors: Martinez
authors: National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, 20850, United States of America.
authors: Ganesha
authors: G
authors: Rai
authors: National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, 20850, United States of America.
authors: Adam
authors: A
authors: Yasgar
authors: National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, 20850, United States of America.
authors: Wendy A
authors: WA
authors: Lea
authors: National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, 20850, United States of America.
authors: Hongmao
authors: H
authors: Sun
authors: National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, 20850, United States of America.
authors: Yuhong
authors: Y
authors: Wang
authors: National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, 20850, United States of America.
authors: Diane K
authors: DK
authors: Luci
authors: National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, 20850, United States of America.
authors: Shyh-Ming
authors: SM
authors: Yang
authors: National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, 20850, United States of America.|Department of Radiation Genetics, Graduate School of Medicine, Kyoto University, Yoshidakonoe, Sakyo, Kyoto 606-8501, Japan.
authors: Kana
authors: K
authors: Nishihara
authors: Department of Neurosurgery, John Hopkins University, Baltimore, MD 21231, United States of America.
authors: Shunichi
authors: S
authors: Takeda
authors: Department of Radiation Genetics, Graduate School of Medicine, Kyoto University, Yoshidakonoe, Sakyo, Kyoto 606-8501, Japan.
authors: Mohiuddin
authors: M
authors: Sagor
authors: Department of Radiation Genetics, Graduate School of Medicine, Kyoto University, Yoshidakonoe, Sakyo, Kyoto 606-8501, Japan.
authors: Irina
authors: I
authors: Earnshaw
authors: Department of Biophysics, Graduate School of Science, Kyoto University, Kitashirakawa-Oiwake, Sakyo, Kyoto 606-8502, Japan.
authors: Tetsuya
authors: T
authors: Okada
authors: Department of Biophysics, Graduate School of Science, Kyoto University, Kitashirakawa-Oiwake, Sakyo, Kyoto 606-8502, Japan.
authors: Kazutoshi
authors: K
authors: Mori
authors: National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, 20850, United States of America.|Department of Neurosurgery, John Hopkins University, Baltimore, MD 21231, United States of America.
authors: Kelli
authors: K
authors: Wilson
authors: Department of Neurosurgery, John Hopkins University, Baltimore, MD 21231, United States of America.
authors: Gregory J
authors: GJ
authors: Riggins
authors: National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, 20850, United States of America.
authors: Menghang
authors: M
authors: Xia
authors: Laboratory of Neuropharmacology, Department of Biochemistry and Molecular Biology, Southern Research Institute, Birmingham, AL 35205, United States of America.
authors: Maurizio
authors: M
authors: Grimaldi
authors: National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, 20850, United States of America.
authors: Ajit
authors: A
authors: Jadhav
authors: National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, 20850, United States of America.
authors: David J
authors: DJ
authors: Maloney
authors: National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, 20850, United States of America.
authors: Anton
authors: A
authors: Simeonov
mesh_terms: D000473:Alkanes / Q000737:chemistry* / Q000494:pharmacology*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001681:Biological Assay / Q000379:methods; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism; D045744:Cell Line, Tumor; D000091342:Endoplasmic Reticulum Chaperone BiP; D059865:Endoplasmic Reticulum Stress / Q000187:drug effects*; D005910:Glioma / Q000378:metabolism*; D019073:HT29 Cells; D006360:Heat-Shock Proteins / Q000378:metabolism; D006801:Humans; D015398:Signal Transduction / Q000187:drug effects
chemical_list: D000473:Alkanes; D000091342:Endoplasmic Reticulum Chaperone BiP; C000717247:HSPA5 protein, human; D006360:Heat-Shock Proteins; C022884:undecane; D002118:Calcium