Assessment of drug-induced mitochondrial dysfunction via altered cellular respiration and acidification measured in a 96-well platform.

Authors

Nadanaciva, Sashi; Rana, Payal; Beeson, Gyda C; Chen, Denise; Ferrick, David A; Beeson, Craig C; Will, Yvonne

Publication Year 2012
Journal Journal of Bioenergetics and Biomembranes
Chapter
Pages 421-437
Volume 44
Issue 4
Issn
Isbn
PMID 22689143.0
PMCID
DOI 10.1007/s10863-012-9446-z
URL http://dx.doi.org/10.1007/s10863-012-9446-z

High-throughput applicable screens for identifying drug-induced mitochondrial impairment are necessary in the pharmaceutical industry. Hence, we evaluated the XF96 Extracellular Flux Analyzer, a 96-well platform that measures changes in the oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) of cells. The sensitivity of the platform was bench-marked with known modulators of oxidative phosphorylation and glycolysis. Sixteen therapeutic agents were screened in HepG2 cells for mitochondrial effects. Four of these compounds, thiazolidinediones, were also tested in primary feline cardiomyocytes for cell-type specific effects. We show that the XF96 platform is a robust, sensitive system for analyzing drug-induced mitochondrial impairment in whole cells. We identified changes in cellular respiration and acidification upon addition of therapeutic agents reported to have a mitochondrial effect. Furthermore, we show that respiration and acidification changes upon addition of the thiazoldinediones were cell-type specific, with the rank order of mitochondrial impairment in whole cells being in accord with the known adverse effects of these drugs.