title: Proteomic characterization of human coronary thrombus in patients with ST-segment elevation acute myocardial infarction.
abstract: Acute myocardial infarction with ST-segment elevation (STEMI) initiates with intraluminal thrombosis and results in total occlusion of the coronary artery. To date, characterization of the coronary thrombus proteome in STEMI patients has not been yet accomplished. Therefore, we aimed to perform an in-depth proteomic characterization of the human coronary thrombus by means of three different approaches: 2-DE followed by mass spectrometry (MALDI MS/MS), 1-DE combined either with liquid chromatography coupled to mass spectrometry in a MALDI TOF/TOF (LC-MALDI-MS/MS), or in a LTQ-Orbitrap (LC-ESI-MS/MS). This approach allowed us to identify a total of 708 proteins in the thrombus. Expression in coronary thrombi (n=20) of 14 proteins was verified, and the expression of fibrin and 6 cell markers (platelets, monocytes, neutrophils, eosinophils, T-cells and B-cells) quantified by selected reaction monitoring (SRM). A positive correlation of 5 proteins (fermitin homolog 3, thrombospondin-1, myosin-9, beta parvin and ras-related protein Rap-1b) with CD41 was found, pointing out the potential activation of a focal adhesion pathway within thrombus platelets. DIDO1 protein was found to correlate negatively with thrombus fibrin, and was found up-regulated in the plasma of these STEMI patients, which may constitute a starting point for further analyses in the search for biomarkers of thrombosis.
  
  
  CONCLUSIONS
  The proteomic characterization of the human coronary thrombus may contribute to a better understanding of the mechanisms involved in acute coronary syndrome, and thus pave the road for the identification of new therapeutic targets that may help addressing this and other thrombotic diseases. A novel methodology to characterize thrombus composition and expression of a sub-group of proteins is hereby described, which allowed linking protein expression with cellular and ECM matrix composition of the thrombus. Five proteins (fermitin homolog 3, thrombospondin-1, myosin-9, beta parvin and ras-related protein Rap-1b) co-express within the human coronary thrombus with CD41, pointing out the potential activation of a focal adhesion pathway within thrombus platelets during thrombus formation. Besides, the protein death-inducer obliterator 1, found to be expressed within the human coronary thrombus, has been proved to increase in the plasma of STEMI patients, which constitutes an important starting point for further analyses in the search for biomarkers of thrombosis.
authors: Department of Vascular Physiopathology, Hospital Nacional de Paraplejicos, SESCAM, Toledo, Spain.
authors: Sergio
authors: S
authors: Alonso-Orgaz
authors: Department of Vascular Physiopathology, Hospital Nacional de Paraplejicos, SESCAM, Toledo, Spain.
authors: Rafael
authors: R
authors: Moreno-Luna
authors: Unidad de Proteomica, CNIC, Madrid, Spain.
authors: Juan A
authors: JA
authors: López
authors: Department of Vascular Physiopathology, Hospital Nacional de Paraplejicos, SESCAM, Toledo, Spain.
authors: Felix
authors: F
authors: Gil-Dones
authors: Cardiology Department, Hospital Virgen de la Salud, SESCAM, Toledo, Spain.
authors: Luis R
authors: LR
authors: Padial
authors: Hemodynamics Department, Hospital Virgen de la Salud, SESCAM, Toledo, Spain.
authors: Jose
authors: J
authors: Moreu
authors: Department of Vascular Physiopathology, Hospital Nacional de Paraplejicos, SESCAM, Toledo, Spain. Electronic address: ferdela@sescam.jccm.es.
authors: Fernando
authors: F
authors: de la Cuesta
authors: Department of Vascular Physiopathology, Hospital Nacional de Paraplejicos, SESCAM, Toledo, Spain.
authors: Maria G
authors: MG
authors: Barderas
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D015415:Biomarkers / Q000378:metabolism; D003328:Coronary Thrombosis / Q000378:metabolism* / Q000473:pathology / Q000601:surgery; D005260:Female; D005786:Gene Expression Regulation; D006801:Humans; D008297:Male; D008875:Middle Aged; D009203:Myocardial Infarction / Q000378:metabolism* / Q000473:pathology / Q000601:surgery; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics
chemical_list: D015415:Biomarkers; D020543:Proteome
keywords: Acute myocardial infarction; DIDO1; Focal adhesion; Human coronary artery; Thrombosis