title: Drug Library Screening for the Identification of Ionophores That Correct the Mistrafficking Disorder Associated with Oxalosis Kidney Disease.
abstract: Primary hyperoxaluria is the underlying cause of oxalosis and is a life-threatening autosomal recessive disease, for which treatment may require dialysis or dual liver-kidney transplantation. The most common primary hyperoxaluria type 1 (PH1) is caused by genetic mutations of a liver-specific enzyme alanine:glyoxylate aminotransferase (AGT), which results in the misrouting of AGT from the peroxisomes to the mitochondria. Pharmacoperones are small molecules with the ability to modify misfolded proteins and route them correctly within the cells, which may present an effective strategy to treat AGT misrouting in PH1 disorders. We miniaturized a cell-based high-content assay into 1536-well plate format and screened ~4200 pharmacologically relevant compounds including Food and Drug Administration, European Union, and Japanese-approved drugs. This assay employs CHO cells stably expressing AGT-170, a mutant that predominantly resides in the mitochondria, where we monitor for its relocation to the peroxisomes through automated image acquisition and analysis. The miniaturized 1536-well assay yielded a Z' averaging 0.70 ± 0.07. Three drugs were identified as potential pharmacoperones from this pilot screen, demonstrating the applicability of this assay for large-scale high-throughput screening.
authors.affiliation: 1 Department of Molecular Therapeutics, Scripps Research Institute Molecular Screening Center, Scripps Research Institute, Jupiter, FL, USA.
authors.forename: Shurong
authors.initials: S
authors.lastname: Hou
authors.affiliation: 1 Department of Molecular Therapeutics, Scripps Research Institute Molecular Screening Center, Scripps Research Institute, Jupiter, FL, USA.|3 Amgen Inc., Thousand Oaks, CA.
authors.forename: Franck
authors.initials: F
authors.lastname: Madoux
authors.affiliation: 1 Department of Molecular Therapeutics, Scripps Research Institute Molecular Screening Center, Scripps Research Institute, Jupiter, FL, USA.
authors.forename: Louis
authors.initials: L
authors.lastname: Scampavia
authors.affiliation: 2 Departments of Internal Medicine and Cell Biology/Biochemistry, Center for Membrane Protein Research, Texas Tech University Health Sciences Center, Lubbock, TX, USA.
authors.forename: Jo Ann
authors.initials: JA
authors.lastname: Janovick
authors.affiliation: 2 Departments of Internal Medicine and Cell Biology/Biochemistry, Center for Membrane Protein Research, Texas Tech University Health Sciences Center, Lubbock, TX, USA.
authors.forename: P Michael
authors.initials: PM
authors.lastname: Conn
authors.affiliation: 1 Department of Molecular Therapeutics, Scripps Research Institute Molecular Screening Center, Scripps Research Institute, Jupiter, FL, USA.
authors.forename: Timothy P
authors.initials: TP
authors.lastname: Spicer
mesh_terms: D000818:Animals; D016466:CHO Cells; D003412:Cricetulus; D004353:Drug Evaluation, Preclinical / Q000379:methods; D006959:Hyperoxaluria / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism; D006960:Hyperoxaluria, Primary / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism; D007476:Ionophores / Q000494:pharmacology*; D007674:Kidney Diseases / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism; D016030:Kidney Transplantation / Q000379:methods; D008099:Liver / Q000187:drug effects / Q000378:metabolism; D008928:Mitochondria / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D020675:Peroxisomes / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D006435:Renal Dialysis / Q000379:methods; D000637:Transaminases / Q000235:genetics / Q000378:metabolism
chemical_list: D007476:Ionophores; D000637:Transaminases; C023067:glyoxylate aminotransferase
keywords: AGT; HCS; oxalosis; pharmacoperone; trafficking