title: Chemical library screening identifies a small molecule that downregulates SOD1 transcription for drugs to treat amyotrophic lateral sclerosis.
abstract: Familial amyotrophic lateral sclerosis (fALS) accounts for 10% of ALS cases, and about 25% of fALS cases are due to mutations in superoxide dismutase 1 (SOD1). Mutant SOD1-mediated ALS is caused by a gain of toxic function of the mutant protein, and the SOD1 level in nonneuronal neighbors, including astrocytes, determines the progression of ALS (non-cell-autonomous toxicity). Therefore, the authors hypothesized that small molecules that reduce SOD1 protein levels in astrocytes might slow the progression of mutant SOD1-mediated ALS. They developed and optimized a cell-based, high-throughput assay to identify low molecular weight compounds that decrease SOD1 expression transcriptionally in human astrocyte-derived cells. Screening of a chemical library of 9600 compounds with the assay identified two hit compounds that selectively and partially downregulate SOD1 expression in a dose-dependent manner, without any detectable cellular toxicity. Western blot analysis showed that one hit compound significantly decreased the level of endogenous SOD1 protein in H4 cells, with no reduction in expression of β-actin. The assay developed here provides a powerful strategy for discovering novel lead molecules for treating familial SOD1-mediated ALS.
authors.affiliation: Department of Neurology, Graduate School of Medicine, Kyoto University, Kyoto, Japan.
authors.forename: Gaku
authors.initials: G
authors.lastname: Murakami
authors.forename: Haruhisa
authors.initials: H
authors.lastname: Inoue
authors.forename: Kayoko
authors.initials: K
authors.lastname: Tsukita
authors.forename: Yasuyuki
authors.initials: Y
authors.lastname: Asai
authors.forename: Yuji
authors.initials: Y
authors.lastname: Amagai
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Aiba
authors.forename: Hiroki
authors.initials: H
authors.lastname: Shimogawa
authors.forename: Motonari
authors.initials: M
authors.lastname: Uesugi
authors.forename: Norio
authors.initials: N
authors.lastname: Nakatsuji
authors.forename: Ryosuke
authors.initials: R
authors.lastname: Takahashi
mesh_terms: D000690:Amyotrophic Lateral Sclerosis / Q000201:enzymology*; D002460:Cell Line; D015536:Down-Regulation / Q000187:drug effects*; D004353:Drug Evaluation, Preclinical; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects*; D017930:Genes, Reporter; D057166:High-Throughput Screening Assays; D006801:Humans; D011401:Promoter Regions, Genetic / Q000235:genetics; D054852:Small Molecule Libraries / Q000494:pharmacology*; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism*; D000072105:Superoxide Dismutase-1
chemical_list: C000606290:SOD1 protein, human; D054852:Small Molecule Libraries; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1