title: Phosphoproteomic profiling of human myocardial tissues distinguishes ischemic from non-ischemic end stage heart failure.
abstract: The molecular differences between ischemic (IF) and non-ischemic (NIF) heart failure are poorly defined. A better understanding of the molecular differences between these two heart failure etiologies may lead to the development of more effective heart failure therapeutics. In this study extensive proteomic and phosphoproteomic profiles of myocardial tissue from patients diagnosed with IF or NIF were assembled and compared. Proteins extracted from left ventricular sections were proteolyzed and phosphopeptides were enriched using titanium dioxide resin. Gel- and label-free nanoscale capillary liquid chromatography coupled to high resolution accuracy mass tandem mass spectrometry allowed for the quantification of 4,436 peptides (corresponding to 450 proteins) and 823 phosphopeptides (corresponding to 400 proteins) from the unenriched and phospho-enriched fractions, respectively. Protein abundance did not distinguish NIF from IF. In contrast, 37 peptides (corresponding to 26 proteins) exhibited a ≥ 2-fold alteration in phosphorylation state (p<0.05) when comparing IF and NIF. The degree of protein phosphorylation at these 37 sites was specifically dependent upon the heart failure etiology examined. Proteins exhibiting phosphorylation alterations were grouped into functional categories: transcriptional activation/RNA processing; cytoskeleton structure/function; molecular chaperones; cell adhesion/signaling; apoptosis; and energetic/metabolism. Phosphoproteomic analysis demonstrated profound post-translational differences in proteins that are involved in multiple cellular processes between different heart failure phenotypes. Understanding the roles these phosphorylation alterations play in the development of NIF and IF has the potential to generate etiology-specific heart failure therapeutics, which could be more effective than current therapeutics in addressing the growing concern of heart failure.
authors.affiliation: Department of Surgery, Duke University Medical Center, Durham, North Carolina, United States of America.
authors.forename: Matthew A
authors.initials: MA
authors.lastname: Schechter
authors.affiliation: Department of Surgery, Duke University Medical Center, Durham, North Carolina, United States of America.
authors.forename: Michael K H
authors.initials: MK
authors.lastname: Hsieh
authors.affiliation: Department of Surgery, Duke University Medical Center, Durham, North Carolina, United States of America.
authors.forename: Linda W
authors.initials: LW
authors.lastname: Njoroge
authors.affiliation: Duke Proteomics Core, Duke University Medical Center, Durham, North Carolina, United States of America.
authors.forename: J Will
authors.initials: JW
authors.lastname: Thompson
authors.affiliation: Duke Proteomics Core, Duke University Medical Center, Durham, North Carolina, United States of America.
authors.forename: Erik J
authors.initials: EJ
authors.lastname: Soderblom
authors.affiliation: Department of Surgery, Duke University Medical Center, Durham, North Carolina, United States of America.
authors.forename: Bryan J
authors.initials: BJ
authors.lastname: Feger
authors.affiliation: Department of Surgery, Duke University Medical Center, Durham, North Carolina, United States of America.
authors.forename: Constantine D
authors.initials: CD
authors.lastname: Troupes
authors.affiliation: Sarah W. Stedman Nutrition and Metabolism Center, Duke University Medical Center, Durham, North Carolina, United States of America; Department of Medicine, Duke University Medical Center, Durham, North Carolina, United States of America.
authors.forename: Kathleen A
authors.initials: KA
authors.lastname: Hershberger
authors.affiliation: Sarah W. Stedman Nutrition and Metabolism Center, Duke University Medical Center, Durham, North Carolina, United States of America.
authors.forename: Olga R
authors.initials: OR
authors.lastname: Ilkayeva
authors.affiliation: Department of Surgery, Duke University Medical Center, Durham, North Carolina, United States of America.
authors.forename: Whitney L
authors.initials: WL
authors.lastname: Nagel
authors.affiliation: Department of Surgery, Duke University Medical Center, Durham, North Carolina, United States of America.
authors.forename: Gina P
authors.initials: GP
authors.lastname: Landinez
authors.affiliation: Department of Surgery, Duke University Medical Center, Durham, North Carolina, United States of America.
authors.forename: Kishan M
authors.initials: KM
authors.lastname: Shah
authors.affiliation: Duke Translational Research Institute, Duke University Medical Center, Durham, North Carolina, United States of America.
authors.forename: Virginia A
authors.initials: VA
authors.lastname: Burns
authors.affiliation: Department of Surgery, Duke University Medical Center, Durham, North Carolina, United States of America.
authors.forename: Lucia
authors.initials: L
authors.lastname: Santacruz
authors.affiliation: Sarah W. Stedman Nutrition and Metabolism Center, Duke University Medical Center, Durham, North Carolina, United States of America; Department of Medicine, Duke University Medical Center, Durham, North Carolina, United States of America.
authors.forename: Matthew D
authors.initials: MD
authors.lastname: Hirschey
authors.affiliation: Division of Pulmonary, Allergy and Critical Care, Medicine, Duke University Medical Center, Durham, North Carolina, United States of America.
authors.forename: Matthew W
authors.initials: MW
authors.lastname: Foster
authors.affiliation: Department of Surgery, Duke University Medical Center, Durham, North Carolina, United States of America.
authors.forename: Carmelo A
authors.initials: CA
authors.lastname: Milano
authors.affiliation: Duke Proteomics Core, Duke University Medical Center, Durham, North Carolina, United States of America.
authors.forename: M Arthur
authors.initials: MA
authors.lastname: Moseley
authors.affiliation: Department of Surgery, Duke University Medical Center, Durham, North Carolina, United States of America.
authors.forename: Valentino
authors.initials: V
authors.lastname: Piacentino
authors.affiliation: Department of Surgery, Duke University Medical Center, Durham, North Carolina, United States of America.
authors.forename: Dawn E
authors.initials: DE
authors.lastname: Bowles
mesh_terms: D000368:Aged; D016000:Cluster Analysis; D019295:Computational Biology; D003937:Diagnosis, Differential; D020869:Gene Expression Profiling; D006333:Heart Failure / Q000175:diagnosis / Q000209:etiology* / Q000378:metabolism*; D006352:Heart Ventricles / Q000378:metabolism; D006801:Humans; D008297:Male; D055442:Metabolome; D055432:Metabolomics; D008875:Middle Aged; D017202:Myocardial Ischemia / Q000150:complications; D009206:Myocardium / Q000378:metabolism*; D010748:Phosphopeptides / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism*; D025941:Protein Interaction Mapping; D060066:Protein Interaction Maps; D020543:Proteome; D040901:Proteomics / Q000379:methods; D015203:Reproducibility of Results
chemical_list: D010748:Phosphopeptides; D010750:Phosphoproteins; D020543:Proteome