title: Metabolic profiles show specific mitochondrial toxicities in vitro in myotube cells.
abstract: Mitochondrial toxicity has been a serious concern, not only in preclinical drug development but also in clinical trials. In mitochondria, there are several distinct metabolic processes including fatty acid β-oxidation, the tricarboxylic acid (TCA) cycle, and oxidative phosphorylation (OXPHOS), and each process contains discrete but often intimately linked steps. Interruption in any one of those steps can cause mitochondrial dysfunction. Detection of inhibition to OXPHOS can be complicated in vivo because intermediate endogenous metabolites can be recycled in situ or circulated systemically for metabolism in other organs or tissues. Commonly used assays for evaluating mitochondrial function are often applied to ex vivo or in vitro samples; they include various enzymatic or protein assays, as well as functional assays such as measurement of oxygen consumption rate, membrane potential, or acidification rates. Metabolomics provides quantitative profiles of overall metabolic changes that can aid in the unraveling of explicit biochemical details of mitochondrial inhibition while providing a holistic view and heuristic understanding of cellular bioenergetics. In this paper, we showed the application of quantitative NMR metabolomics to in vitro myotube cells treated with mitochondrial toxicants, rotenone and antimycin A. The close coupling of the TCA cycle to the electron transfer chain (ETC) in OXPHOS enables specific diagnoses of inhibition to ETC complexes by discrete biochemical changes in the TCA cycle.
authors.affiliation: Merck Research Laboratories, 770 Sumneytown Pike, West Point, PA 19486, USA. qiuwei_xu@merck.com
authors.forename: Qiuwei
authors.initials: Q
authors.lastname: Xu
authors.forename: Heather
authors.initials: H
authors.lastname: Vu
authors.forename: Liping
authors.initials: L
authors.lastname: Liu
authors.forename: Ting-Chuan
authors.initials: TC
authors.lastname: Wang
authors.forename: William H
authors.initials: WH
authors.lastname: Schaefer
mesh_terms: D000968:Antimycin A / Q000633:toxicity; D002478:Cells, Cultured; D002952:Citric Acid Cycle / Q000187:drug effects; D004579:Electron Transport / Q000187:drug effects; D006801:Humans; D055442:Metabolome / Q000187:drug effects*; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism*; D018485:Muscle Fibers, Skeletal / Q000473:pathology*; D010085:Oxidative Phosphorylation / Q000187:drug effects*; D012402:Rotenone / Q000633:toxicity; D014475:Uncoupling Agents / Q000494:pharmacology* / Q000633:toxicity
chemical_list: D014475:Uncoupling Agents; D012402:Rotenone; D000968:Antimycin A