title: Cardiac phosphoproteome reveals cell signaling events involved in doxorubicin cardiotoxicity.
abstract: The successful use of anthracyclines like doxorubicin in chemotherapy is limited by their severe cardiotoxicity. Despite decades of clinical application, a satisfying description of the molecular mechanisms involved and a preventive treatment have not yet been achieved. Here we address doxorubicin-induced changes in cell signaling as a novel potential mediator of doxorubicin toxicity by applying a non-biased screen of the cardiac phosphoproteome. Two-dimensional gel electrophoresis, phosphospecific staining, quantitative image analysis, and MALDI-TOF/TOF mass spectrometry were combined to identify (de)phosphorylation events occurring in the isolated rat heart upon Langendorff-perfusion with clinically relevant (5 μM) and supraclinical concentrations (25 μM) of doxorubicin. This approach identified 22 proteins with a significantly changed phosphorylation status and these results were validated by immunoblotting for selected phosphosites. Overrepresentation of mitochondrial proteins (>40%) identified this compartment as a prime target of doxorubicin. Identified proteins were mainly involved in energy metabolism (e.g. pyruvate dehydrogenase and acyl-CoA dehydrogenase), sarcomere structure and function (e.g. desmin) or chaperone-like activities (e.g. α-crystallin B chain and prohibitin). Changes in phosphorylation of pyruvate dehydrogenase, regulating pyruvate entry into the Krebs cycle, and desmin, maintaining myofibrillar array, are relevant for main symptoms of cardiac dysfunction related to doxorubicin treatment, namely energy imbalance and myofibrillar disorganization. This article is part of a Special Issue entitled: Translational Proteomics.
authors.affiliation: University Joseph Fourier, Laboratory of Fundamental and Applied Bioenergetics, Environmental and Systems Biology, Grenoble, France.
authors.forename: Séverine
authors.initials: S
authors.lastname: Gratia
authors.forename: Laurence
authors.initials: L
authors.lastname: Kay
authors.forename: Sylvie
authors.initials: S
authors.lastname: Michelland
authors.forename: Michel
authors.initials: M
authors.lastname: Sève
authors.forename: Uwe
authors.initials: U
authors.lastname: Schlattner
authors.forename: Malgorzata
authors.initials: M
authors.lastname: Tokarska-Schlattner
mesh_terms: D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000009:adverse effects* / Q000494:pharmacology; D004317:Doxorubicin / Q000009:adverse effects* / Q000494:pharmacology; D006331:Heart Diseases / Q000139:chemically induced / Q000378:metabolism* / Q000473:pathology; D008297:Male; D009124:Muscle Proteins / Q000378:metabolism*; D009206:Myocardium / Q000378:metabolism* / Q000473:pathology; D010766:Phosphorylation / Q000187:drug effects; D020543:Proteome / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D015398:Signal Transduction / Q000187:drug effects*
chemical_list: D000903:Antibiotics, Antineoplastic; D009124:Muscle Proteins; D020543:Proteome; D004317:Doxorubicin