title: Antibody phage display assisted identification of junction plakoglobin as a potential biomarker for atherosclerosis.
abstract: To date, no plaque-derived blood biomarker is available to allow diagnosis, prognosis or monitoring of atherosclerotic vascular diseases. In this study, specimens of thrombendarterectomy material from carotid and iliac arteries were incubated in protein-free medium to obtain plaque and control secretomes for subsequent subtractive phage display. The selection of nine plaque secretome-specific antibodies and the analysis of their immunopurified antigens by mass spectrometry led to the identification of 22 proteins. One of them, junction plakoglobin (JUP-81) and its smaller isoforms (referred to as JUP-63, JUP-55 and JUP-30 by molecular weight) were confirmed by immunohistochemistry and immunoblotting with independent antibodies to be present in atherosclerotic plaques and their secretomes, coronary thrombi of patients with acute coronary syndrome (ACS) and macrophages differentiated from peripheral blood monocytes as well as macrophage-like cells differentiated from THP1 cells. Plasma of patients with stable coronary artery disease (CAD) (n = 15) and ACS (n = 11) contained JUP-81 at more than 2- and 14-fold higher median concentrations, respectively, than plasma of CAD-free individuals (n = 13). In conclusion, this proof of principle study identified and verified JUP isoforms as potential plasma biomarkers for atherosclerosis. Clinical validation studies are needed to determine its diagnostic efficacy and clinical utility as a biomarker for diagnosis, prognosis or monitoring of atherosclerotic vascular diseases.
authors.affiliation: Institute for Clinical Chemistry, University Hospital Zurich, Zurich, Switzerland.
authors.forename: Seraina
authors.initials: S
authors.lastname: Cooksley-Decasper
authors.forename: Hans
authors.initials: H
authors.lastname: Reiser
authors.forename: Daniela S
authors.initials: DS
authors.lastname: Thommen
authors.forename: Barbara
authors.initials: B
authors.lastname: Biedermann
authors.forename: Michel
authors.initials: M
authors.lastname: Neidhart
authors.forename: Joanna
authors.initials: J
authors.lastname: Gawinecka
authors.forename: Gieri
authors.initials: G
authors.lastname: Cathomas
authors.forename: Fabian C
authors.initials: FC
authors.lastname: Franzeck
authors.forename: Christophe
authors.initials: C
authors.lastname: Wyss
authors.forename: Roland
authors.initials: R
authors.lastname: Klingenberg
authors.forename: Paolo
authors.initials: P
authors.lastname: Nanni
authors.forename: Bernd
authors.initials: B
authors.lastname: Roschitzki
authors.forename: Christian
authors.initials: C
authors.lastname: Matter
authors.forename: Petra
authors.initials: P
authors.lastname: Wolint
authors.forename: Maximilian Y
authors.initials: MY
authors.lastname: Emmert
authors.forename: Marc
authors.initials: M
authors.lastname: Husmann
authors.forename: Beatrice
authors.initials: B
authors.lastname: Amann-Vesti
authors.forename: Wilibald
authors.initials: W
authors.lastname: Maier
authors.forename: Steffen
authors.initials: S
authors.lastname: Gay
authors.forename: Thomas F
authors.initials: TF
authors.lastname: Lüscher
authors.forename: Arnold
authors.initials: A
authors.lastname: von Eckardstein
authors.forename: Danielle
authors.initials: D
authors.lastname: Hof
mesh_terms: D054058:Acute Coronary Syndrome / Q000097:blood / Q000378:metabolism; D000368:Aged; D000595:Amino Acid Sequence; D000906:Antibodies / Q000235:genetics / Q000378:metabolism*; D050197:Atherosclerosis / Q000175:diagnosis / Q000378:metabolism*; D015415:Biomarkers / Q000097:blood / Q000378:metabolism*; D002339:Carotid Arteries / Q000378:metabolism / Q000473:pathology / Q000601:surgery; D002460:Cell Line; D003324:Coronary Artery Disease / Q000097:blood / Q000378:metabolism; D051180:Desmoplakins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D004691:Endarterectomy; D006801:Humans; D007083:Iliac Artery / Q000378:metabolism / Q000473:pathology / Q000601:surgery; D015151:Immunoblotting; D007150:Immunohistochemistry; D008264:Macrophages / Q000378:metabolism / Q000473:pathology; D008297:Male; D013058:Mass Spectrometry; D008875:Middle Aged; D008969:Molecular Sequence Data; D019151:Peptide Library; D058226:Plaque, Atherosclerotic / Q000097:blood / Q000378:metabolism; D020033:Protein Isoforms / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D040901:Proteomics / Q000379:methods; D017386:Sequence Homology, Amino Acid; D051185:gamma Catenin
chemical_list: D000906:Antibodies; D015415:Biomarkers; D051180:Desmoplakins; C495292:JUP protein, human; D019151:Peptide Library; D020033:Protein Isoforms; D051185:gamma Catenin