title: A high-content screen identifies novel compounds that inhibit stress-induced TDP-43 cellular aggregation and associated cytotoxicity.
abstract: TDP-43 is an RNA binding protein found to accumulate in the cytoplasm of brain and spinal cord from patients affected with amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). Nuclear TDP-43 protein regulates transcription through several mechanisms, and under stressed conditions, it forms cytoplasmic aggregates that co-localize with stress granule (SG) proteins in cell culture. These granules are also found in the brain and spinal cord of patients affected with ALS and FTLD. The mechanism through which TDP-43 might contribute to neurodegenerative diseases is poorly understood. To investigate the pathophysiology of TDP-43 aggregation and to isolate potential therapeutic targets, we screened a chemical library of 75,000 compounds using high-content analysis with PC12 cells that inducibly express human TDP-43 tagged with green fluorescent protein (GFP). The screen identified 16 compounds that dose-dependently decreased the TDP-43 inclusions without significant cellular toxicity or changes in total TDP-43 expression levels. To validate the effect, we tested compounds by Western blot analysis and in a Caenorhabditis elegans model that replicates some of the relevant disease phenotypes. The hits from this assay will be useful for elucidating regulation of TDP-43, stress granule response, and possible ALS therapeutics.
authors.affiliation: Laboratory for Drug Discovery in Neurodegeneration, Harvard NeuroDiscovery Center, Brigham and Women's Hospital and Harvard Medical School, Cambridge, MA.
authors.forename: Justin D
authors.initials: JD
authors.lastname: Boyd
authors.affiliation: Department of Pharmacology and Experimental Therapeutics, Boston University School of Medicine, Boston, MA.
authors.forename: Peter
authors.initials: P
authors.lastname: Lee
authors.affiliation: Department of Pharmacology and Experimental Therapeutics, Boston University School of Medicine, Boston, MA.
authors.forename: Marisa S
authors.initials: MS
authors.lastname: Feiler
authors.affiliation: Department of Pharmacology and Experimental Therapeutics, Boston University School of Medicine, Boston, MA.
authors.forename: Nava
authors.initials: N
authors.lastname: Zauur
authors.affiliation: Laboratory for Drug Discovery in Neurodegeneration, Harvard NeuroDiscovery Center, Brigham and Women's Hospital and Harvard Medical School, Cambridge, MA.
authors.forename: Min
authors.initials: M
authors.lastname: Liu
authors.affiliation: Laboratory for Drug Discovery in Neurodegeneration, Harvard NeuroDiscovery Center, Brigham and Women's Hospital and Harvard Medical School, Cambridge, MA.
authors.forename: John
authors.initials: J
authors.lastname: Concannon
authors.affiliation: Department of Pharmacology and Experimental Therapeutics, Boston University School of Medicine, Boston, MA.|Department of Pathology and Laboratory Medicine, Boston University School of Medicine, Boston, MA.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Ebata
authors.affiliation: Department of Pharmacology and Experimental Therapeutics, Boston University School of Medicine, Boston, MA.|Department of Neurology, Boston University School of Medicine, Boston, MA.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Wolozin
authors.affiliation: Laboratory for Drug Discovery in Neurodegeneration, Harvard NeuroDiscovery Center, Brigham and Women's Hospital and Harvard Medical School, Cambridge, MA.
authors.forename: Marcie A
authors.initials: MA
authors.lastname: Glicksman
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D018053:Arsenites / Q000494:pharmacology; D017173:Caenorhabditis elegans; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D055808:Drug Discovery / Q000379:methods; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D015870:Gene Expression; D017930:Genes, Reporter; D057166:High-Throughput Screening Assays; D006801:Humans; D019636:Neurodegenerative Diseases / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism; D018696:Neuroprotective Agents / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D054852:Small Molecule Libraries; D017670:Sodium Compounds / Q000494:pharmacology; D013312:Stress, Physiological / Q000187:drug effects*
chemical_list: D018053:Arsenites; D004268:DNA-Binding Proteins; D018696:Neuroprotective Agents; D011993:Recombinant Fusion Proteins; D054852:Small Molecule Libraries; D017670:Sodium Compounds; C017947:sodium arsenite
keywords: RNA binding protein; RNA granule; aggregation; amyotrophic lateral sclerosis; high-throughput screen; protein synthesis
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