title: LC-MS Analysis of Human Platelets as a Platform for Studying Mitochondrial Metabolism.
abstract: Perturbed mitochondrial metabolism has received renewed interest as playing a causative role in a range of diseases. Probing alterations to metabolic pathways requires a model in which external factors can be well controlled, allowing for reproducible and meaningful results. Many studies employ transformed cellular models for these purposes; however, metabolic reprogramming that occurs in many cancer cell lines may introduce confounding variables. For this reason primary cells are desirable, though attaining adequate biomass for metabolic studies can be challenging. Here we show that human platelets can be utilized as a platform to carry out metabolic studies in combination with liquid chromatography-tandem mass spectrometry analysis. This approach is amenable to relative quantification and isotopic labeling to probe the activity of specific metabolic pathways. Availability of platelets from individual donors or from blood banks makes this model system applicable to clinical studies and feasible to scale up. Here we utilize isolated platelets to confirm previously identified compensatory metabolic shifts in response to the complex I inhibitor rotenone. More specifically, a decrease in glycolysis is accompanied by an increase in fatty acid oxidation to maintain acetyl-CoA levels. Our results show that platelets can be used as an easily accessible and medically relevant model to probe the effects of xenobiotics on cellular metabolism.
authors.affiliation: Center for Cancer Pharmacology, University of Pennsylvania; Center for Excellence in Environmental Toxicology, University of Pennsylvania.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Worth
authors.affiliation: Center for Excellence in Environmental Toxicology, University of Pennsylvania; Penn SRP and Department of Systems Pharmacology and Translational Therapeutics, University of Pennsylvania.
authors.forename: Dylan M
authors.initials: DM
authors.lastname: Marchione
authors.affiliation: Center for Cancer Pharmacology, University of Pennsylvania; Center for Excellence in Environmental Toxicology, University of Pennsylvania.
authors.forename: Robert C
authors.initials: RC
authors.lastname: Parry
authors.affiliation: Center for Excellence in Environmental Toxicology, University of Pennsylvania; Penn SRP and Department of Systems Pharmacology and Translational Therapeutics, University of Pennsylvania.
authors.forename: Qingqing
authors.initials: Q
authors.lastname: Wang
authors.affiliation: Center for Excellence in Environmental Toxicology, University of Pennsylvania; Penn SRP and Department of Systems Pharmacology and Translational Therapeutics, University of Pennsylvania.
authors.forename: Kevin P
authors.initials: KP
authors.lastname: Gillespie
authors.affiliation: Division of Traumatology, Department of Surgery, Critical Care and Acute Care Surgery, University of Pennsylvania.
authors.forename: Noelle N
authors.initials: NN
authors.lastname: Saillant
authors.affiliation: Division of Traumatology, Department of Surgery, Critical Care and Acute Care Surgery, University of Pennsylvania.
authors.forename: Carrie
authors.initials: C
authors.lastname: Sims
authors.affiliation: Center for Cancer Pharmacology, University of Pennsylvania; Center for Excellence in Environmental Toxicology, University of Pennsylvania.
authors.forename: Clementina
authors.initials: C
authors.lastname: Mesaros
authors.affiliation: A.J. Drexel Autism Institute, Drexel University.
authors.forename: Nathaniel W
authors.initials: NW
authors.lastname: Snyder
authors.affiliation: Center for Cancer Pharmacology, University of Pennsylvania; Center for Excellence in Environmental Toxicology, University of Pennsylvania; ianblair@mail.med.upenn.edu.
authors.forename: Ian A
authors.initials: IA
authors.lastname: Blair
mesh_terms: D001792:Blood Platelets / Q000378:metabolism*; D002460:Cell Line; D002853:Chromatography, Liquid / Q000379:methods*; D006801:Humans; D007553:Isotope Labeling / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods*; D053858:Metabolic Networks and Pathways; D008928:Mitochondria / Q000378:metabolism*; D010084:Oxidation-Reduction; D012402:Rotenone / Q000494:pharmacology; D014475:Uncoupling Agents / Q000494:pharmacology
chemical_list: D014475:Uncoupling Agents; D012402:Rotenone