title: A cellular genetics approach identifies gene-drug interactions and pinpoints drug toxicity pathway nodes.
abstract: New approaches to toxicity testing have incorporated high-throughput screening across a broad-range of in vitro assays to identify potential key events in response to chemical or drug treatment. To date, these approaches have primarily utilized repurposed drug discovery assays. In this study, we describe an approach that combines in vitro screening with genetic approaches for the experimental identification of genes and pathways involved in chemical or drug toxicity. Primary embryonic fibroblasts isolated from 32 genetically-characterized inbred mouse strains were treated in concentration-response format with 65 compounds, including pharmaceutical drugs, environmental chemicals, and compounds with known modes-of-action. Integrated cellular responses were measured at 24 and 72 h using high-content imaging and included cell loss, membrane permeability, mitochondrial function, and apoptosis. Genetic association analysis of cross-strain differences in the cellular responses resulted in a collection of candidate loci potentially underlying the variable strain response to each chemical. As a demonstration of the approach, one candidate gene involved in rotenone sensitivity, Cybb, was experimentally validated in vitro and in vivo. Pathway analysis on the combined list of candidate loci across all chemicals identified a number of over-connected nodes that may serve as core regulatory points in toxicity pathways.
authors.affiliation: Division of Pharmacotherapy and Experimental Therapeutics, UNC Eshelman School of Pharmacy at the University of North Carolina at Chapel Hill Chapel Hill, NC, USA.
authors.forename: Oscar T
authors.initials: OT
authors.lastname: Suzuki
authors.affiliation: Division of Pharmacotherapy and Experimental Therapeutics, UNC Eshelman School of Pharmacy at the University of North Carolina at Chapel Hill Chapel Hill, NC, USA.
authors.forename: Amber
authors.initials: A
authors.lastname: Frick
authors.affiliation: The Hamner Institutes for Health Sciences, Research Triangle Park NC, USA.
authors.forename: Bethany B
authors.initials: BB
authors.lastname: Parks
authors.affiliation: The Hamner Institutes for Health Sciences, Research Triangle Park NC, USA.
authors.forename: O Joseph
authors.initials: OJ
authors.lastname: Trask
authors.affiliation: Division of Pharmacotherapy and Experimental Therapeutics, UNC Eshelman School of Pharmacy at the University of North Carolina at Chapel Hill Chapel Hill, NC, USA.
authors.forename: Natasha
authors.initials: N
authors.lastname: Butz
authors.affiliation: Division of Pharmacotherapy and Experimental Therapeutics, UNC Eshelman School of Pharmacy at the University of North Carolina at Chapel Hill Chapel Hill, NC, USA.
authors.forename: Brian
authors.initials: B
authors.lastname: Steffy
authors.affiliation: Division of Pharmacotherapy and Experimental Therapeutics, UNC Eshelman School of Pharmacy at the University of North Carolina at Chapel Hill Chapel Hill, NC, USA.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Chan
authors.affiliation: Division of Pharmacotherapy and Experimental Therapeutics, UNC Eshelman School of Pharmacy at the University of North Carolina at Chapel Hill Chapel Hill, NC, USA.
authors.forename: David K
authors.initials: DK
authors.lastname: Scoville
authors.affiliation: The Hamner Institutes for Health Sciences, Research Triangle Park NC, USA.
authors.forename: Eric
authors.initials: E
authors.lastname: Healy
authors.affiliation: Division of Pharmacotherapy and Experimental Therapeutics, UNC Eshelman School of Pharmacy at the University of North Carolina at Chapel Hill Chapel Hill, NC, USA.
authors.forename: Cristina
authors.initials: C
authors.lastname: Benton
authors.affiliation: Meredith College Raleigh, NC, USA.
authors.forename: Patricia E
authors.initials: PE
authors.lastname: McQuaid
authors.affiliation: The Hamner Institutes for Health Sciences, Research Triangle Park NC, USA.
authors.forename: Russell S
authors.initials: RS
authors.lastname: Thomas
authors.affiliation: Division of Pharmacotherapy and Experimental Therapeutics, UNC Eshelman School of Pharmacy at the University of North Carolina at Chapel Hill Chapel Hill, NC, USA.
authors.forename: Tim
authors.initials: T
authors.lastname: Wiltshire
keywords: gene-drug interaction; genome-wide association; high throughput screening; high-content imaging; in vitro screening; mouse model