title: Identification of a small molecule that induces ATG5-and-cathepsin-l-dependent cell death and modulates polyglutamine toxicity.
abstract: Non-apoptotic cell death mechanisms are largely uncharacterized despite their importance in physiology and disease [1]. Here we sought to systematically identify non-apoptotic cell death pathways in mammalian cells. We screened 69,612 compounds for those that induce non-canonical cell death by counter screening in the presence of inhibitors of apoptosis and necrosis. We further selected compounds that require active protein synthesis for inducing cell death. Using this tiered approach, we identified NID-1 (Novel Inducer of Death-1), a small molecule that induces an active, energy-dependent cell death in diverse mammalian cell lines. NID-1-induced death required components of the autophagic machinery, including ATG5, and the lysosomal hydrolase cathepsin L, but was distinct from classical macroautophagy. Since macroautophagy can prevent cell death in several contexts, we tested and found that NID-1 suppressed cell death in a cell-based model of Huntington's disease, suggesting that NID-1 activates a specific pathway. Thus the discovery of NID-1 identifies a previously unexplored cell death pathway, and modulating this pathway may have therapeutic applications. Furthermore, these findings provide a proof-of-principle for using chemical screening to identify novel cell death paradigms.
authors.affiliation: Howard Hughes Medical Institute, Department of Biological Sciences, Columbia University, Northwest Corner Building, MC 4846, 550 West 120th Street, New York, NY 10027, United States; Department of Pathology and Cell Biology, Columbia University Medical Center, New York, NY 10032, United States.
authors.forename: Hemant
authors.initials: H
authors.lastname: Varma
authors.affiliation: Howard Hughes Medical Institute, Department of Biological Sciences, Columbia University, Northwest Corner Building, MC 4846, 550 West 120th Street, New York, NY 10027, United States.
authors.forename: Nidhi M
authors.initials: NM
authors.lastname: Gangadhar
authors.affiliation: Howard Hughes Medical Institute, Department of Biological Sciences, Columbia University, Northwest Corner Building, MC 4846, 550 West 120th Street, New York, NY 10027, United States.
authors.forename: Reka R
authors.initials: RR
authors.lastname: Letso
authors.affiliation: Howard Hughes Medical Institute, Department of Biological Sciences, Columbia University, Northwest Corner Building, MC 4846, 550 West 120th Street, New York, NY 10027, United States.
authors.forename: Adam J
authors.initials: AJ
authors.lastname: Wolpaw
authors.affiliation: Howard Hughes Medical Institute, Department of Biological Sciences, Columbia University, Northwest Corner Building, MC 4846, 550 West 120th Street, New York, NY 10027, United States.
authors.forename: Rohitha
authors.initials: R
authors.lastname: Sriramaratnam
authors.affiliation: Howard Hughes Medical Institute, Department of Biological Sciences, Columbia University, Northwest Corner Building, MC 4846, 550 West 120th Street, New York, NY 10027, United States; Department of Chemistry, Columbia University, Northwest Corner Building, MC 4846, 550 West 120th Street, New York, NY 10027, United States. Electronic address: bstockwell@columbia.edu.
authors.forename: Brent R
authors.initials: BR
authors.lastname: Stockwell
mesh_terms: D000818:Animals; D017209:Apoptosis; D001343:Autophagy / Q000187:drug effects*; D000071187:Autophagy-Related Protein 5; D056668:Cathepsin L / Q000378:metabolism*; D057166:High-Throughput Screening Assays; D000071058:Huntingtin Protein; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D009336:Necrosis; D009419:Nerve Tissue Proteins / Q000187:drug effects / Q000378:metabolism; D009687:Nuclear Proteins / Q000187:drug effects / Q000378:metabolism; D016716:PC12 Cells; D010455:Peptides / Q000633:toxicity; D014176:Protein Biosynthesis / Q000187:drug effects; D051381:Rats; D054852:Small Molecule Libraries / Q000032:analysis / Q000494:pharmacology; D013876:Thiophenes / Q000494:pharmacology*
chemical_list: C495773:Atg5 protein, mouse; D000071187:Autophagy-Related Protein 5; C499524:Htt protein, rat; D000071058:Huntingtin Protein; D008869:Microtubule-Associated Proteins; C583289:NID-1 compound; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; D010455:Peptides; D054852:Small Molecule Libraries; D013876:Thiophenes; C097188:polyglutamine; D056668:Cathepsin L