title: Development of a phenotypic high-content assay to identify pharmacoperone drugs for the treatment of primary hyperoxaluria type 1 by high-throughput screening.
abstract: Primary hyperoxaluria is a severe disease for which the best current therapy is dialysis or organ transplantation. These are risky, inconvenient, and costly procedures. In some patients, pyridoxine treatment can delay the need for these surgical procedures. The underlying cause of particular forms of this disease is the misrouting of a specific enzyme, alanine:glyoxylate aminotransferase (AGT), to the mitochondria instead of the peroxisomes. Pharmacoperones are small molecules that can rescue misfolded proteins and redirect them to their correct location, thereby restoring their function and potentially curing disease. In the present study, we miniaturized a cell-based assay to identify pharmacoperone drugs present in large chemical libraries to selectively correct AGT misrouting. This assay employs AGT-170, a mutant form of AGT that predominantly resides in the mitochondria, which we monitor for its relocation to the peroxisomes through automated image acquisition and analysis. Over the course of a pilot screen of 1,280 test compounds, we achieved an average Z'-factor of 0.72±0.02, demonstrating the suitability of this assay for HTS.
authors.affiliation: 1 Lead Identification Division, Translational Research Institute, Scripps Research Institute , Jupiter, Florida.
authors.forename: Franck
authors.initials: F
authors.lastname: Madoux
authors.forename: Jo Ann
authors.initials: JA
authors.lastname: Janovick
authors.forename: David
authors.initials: D
authors.lastname: Smithson
authors.forename: Sonia
authors.initials: S
authors.lastname: Fargue
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Danpure
authors.forename: Louis
authors.initials: L
authors.lastname: Scampavia
authors.forename: Yih-Tai
authors.initials: YT
authors.lastname: Chen
authors.forename: Timothy P
authors.initials: TP
authors.lastname: Spicer
authors.forename: P Michael
authors.initials: PM
authors.lastname: Conn
mesh_terms: D000818:Animals; D001681:Biological Assay / Q000379:methods*; D016466:CHO Cells; D002470:Cell Survival / Q000187:drug effects; D003412:Cricetulus; D015195:Drug Design; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D006960:Hyperoxaluria, Primary / Q000188:drug therapy* / Q000473:pathology*; D018832:Molecular Chaperones / Q000138:chemical synthesis / Q000145:classification / Q000494:pharmacology*; D010641:Phenotype; D013678:Technology, Pharmaceutical / Q000379:methods
chemical_list: D018832:Molecular Chaperones