title: A Chemogenomic Screening Platform Used to Identify Chemotypes Perturbing HSP90 Pathways.
abstract: Compounds that modulate the heat shock protein (HSP) network have potential in a broad range of research applications and diseases. A yeast-based liquid culture assay that measured time-dependent turbidity enabled the high-throughput screening of different Saccharomyces cerevisae strains to identify HSP modulators with unique molecular mechanisms. A focused set of four strains, with differing sensitivities to Hsp90 inhibitors, was used to screen a compound library of 3680 compounds. Computed turbidity curve functions were used to classify strain responses and sensitivity to chemical effects across the compound library. Filtering based on single-strain selectivity identified nine compounds as potential heat shock modulators, including the known Hsp90 inhibitor macbecin. Haploid yeast deletion strains (360), mined from previous Hsp90 inhibitor yeast screens and heat shock protein interaction data, were screened for differential sensitivities to known N-terminal ATP site-directed Hsp90 inhibitors to reveal functional distinctions. Strains demonstrating differential sensitivity (13) to Hsp90 inhibitors were used to prioritize primary screen hit compounds, with NSC145366 emerging as the lead hit. Our follow-up biochemical and functional studies show that NSC145366 directly interacts and inhibits the C-terminus of Hsp90, validating the platform as a powerful approach for early-stage identification of bioactive modulators of heat shock-dependent pathways.
authors.affiliation: 1 Department of Medicinal Chemistry and Molecular Pharmacology, College of Pharmacy, Purdue University, West Lafayette, IN, USA.
authors.forename: Fiona M
authors.initials: FM
authors.lastname: Thomas
authors.affiliation: 1 Department of Medicinal Chemistry and Molecular Pharmacology, College of Pharmacy, Purdue University, West Lafayette, IN, USA.
authors.forename: Kourtney M
authors.initials: KM
authors.lastname: Goode
authors.affiliation: 2 Bindley Bioscience Center, Purdue Discovery Park, Purdue University, West Lafayette, IN, USA.
authors.forename: Bartek
authors.initials: B
authors.lastname: Rajwa
authors.affiliation: 1 Department of Medicinal Chemistry and Molecular Pharmacology, College of Pharmacy, Purdue University, West Lafayette, IN, USA.
authors.forename: Andrew A
authors.initials: AA
authors.lastname: Bieberich
authors.affiliation: 2 Bindley Bioscience Center, Purdue Discovery Park, Purdue University, West Lafayette, IN, USA.
authors.forename: Larisa V
authors.initials: LV
authors.lastname: Avramova
authors.affiliation: 1 Department of Medicinal Chemistry and Molecular Pharmacology, College of Pharmacy, Purdue University, West Lafayette, IN, USA.|3 Purdue University Center for Cancer Research, West Lafayette, IN, USA.
authors.forename: Tony R
authors.initials: TR
authors.lastname: Hazbun
authors.affiliation: 1 Department of Medicinal Chemistry and Molecular Pharmacology, College of Pharmacy, Purdue University, West Lafayette, IN, USA.|3 Purdue University Center for Cancer Research, West Lafayette, IN, USA.
authors.forename: V Jo
authors.initials: VJ
authors.lastname: Davisson
mesh_terms: D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D055808:Drug Discovery; D004354:Drug Screening Assays, Antitumor / Q000379:methods; D018841:HSP90 Heat-Shock Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006238:Haploidy; D057166:High-Throughput Screening Assays; D006801:Humans; D015394:Molecular Structure; D010641:Phenotype; D017384:Sequence Deletion; D015398:Signal Transduction / Q000187:drug effects*; D054852:Small Molecule Libraries; D013329:Structure-Activity Relationship
chemical_list: D018841:HSP90 Heat-Shock Proteins; D054852:Small Molecule Libraries
keywords: Hsp90 C-terminal inhibitor; heat shock protein 90; high-throughput assay; phenotypic screen; target identification