title: Kindlin 3 (FERMT3) is associated with unstable atherosclerotic plaques, anti-inflammatory type II macrophages and upregulation of beta-2 integrins in all major arterial beds.
abstract: BACKGROUND
  Kindlins (FERMT) are cytoplasmic proteins required for integrin (ITG) activation, leukocyte transmigration, platelet aggregation and thrombosis. Characterization of kindlins and their association with atherosclerotic plaques in human(s) is lacking.
  
  
  RESULTS
  Exploratory microarray (MA) was first performed followed by selective quantitative validation of robustly expressed genes with qRT-PCR low-density array (LDA). In LDA, ITGA1 (1.30-fold, p = 0.041) and ITGB3 (1.37-fold, p = 0.036) were upregulated in whole blood samples of patients with coronary artery disease (CAD) compared to healthy controls. In arterial plaques, both robustly expressed transcript variants of FERMT3 (MA: 5.90- and 3.4-fold; LDA: 3.99-fold, p < 0.0001 for all) and ITGB2 (MA: 4.81- and 4.92-fold; LDA: 5.29-fold, p < 0.0001 for all) were upregulated while FERMT2 was downregulated (MA: -1.61-fold; LDA: -2.88-fold, p < 0.0001 for both). The other integrins (ITGA1, ITGAV, ITGB3, ITGB5) were downregulated. All these results were replicated in at least one arterial bed. The latter FERMT3 transcript variant associated with unstable plaques (p = 0.0004). FERMT3 correlated with M2 macrophage markers and in hierarchical cluster analysis clustered with inflammatory and macrophage markers, while FERMT2 correlated with SMC-rich plaque markers and clustered with SMC markers. In confocal immunofluorescence analysis, FERMT3 protein colocalized with abundant CD68-positive cells of monocytic origin in the atherosclerotic plaques, while co-localization of FERMT3 with HHF35 indicative of smooth muscle cells was low.
  
  
  CONCLUSIONS
  Kindlin-3 (FERMT3) is upregulated in atherosclerotic, especially unstable plaques, mainly in cells of monocytic origin and of M2 type. Simultaneous upregulation of ITGB2 suggests a synergistic effect on leukocyte adherence and transmigration into the vessel wall.
authors.affiliation: Department of Clinical Chemistry, Fimlab Laboratories, Tampere University Hospital, Tampere, Finland; School of Medicine, University of Tampere, Finland; Division of Vascular Surgery, Department of Surgery, Tampere University Hospital, Finland. Electronic address: nikuoksala@gmail.com.
authors.forename: Niku
authors.initials: N
authors.lastname: Oksala
authors.affiliation: BioMediTech, University of Tampere, Tampere, Finland and Fimlab Laboratories, Tampere University Hospital, Tampere, Finland.
authors.forename: Jenita
authors.initials: J
authors.lastname: Pärssinen
authors.affiliation: Department of Clinical Chemistry, Fimlab Laboratories, Tampere University Hospital, Tampere, Finland.
authors.forename: Ilkka
authors.initials: I
authors.lastname: Seppälä
authors.affiliation: Research Unit of Molecular Epidemiology, Helmholtz Zentrum, German Research Center for Environmental Health, Munich, Germany; Hannover Unified Biobank, Hannover Medical School, Hannover, Germany.
authors.forename: Norman
authors.initials: N
authors.lastname: Klopp
authors.affiliation: Research Unit of Molecular Epidemiology, Helmholtz Zentrum, German Research Center for Environmental Health, Munich, Germany; Hannover Unified Biobank, Hannover Medical School, Hannover, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Illig
authors.affiliation: Department of Clinical Chemistry, Fimlab Laboratories, Tampere University Hospital, Tampere, Finland; School of Medicine, University of Tampere, Finland.
authors.forename: Reijo
authors.initials: R
authors.lastname: Laaksonen
authors.affiliation: Department of Clinical Chemistry, Fimlab Laboratories, Tampere University Hospital, Tampere, Finland.
authors.forename: Mari
authors.initials: M
authors.lastname: Levula
authors.affiliation: Department of Clinical Chemistry, Fimlab Laboratories, Tampere University Hospital, Tampere, Finland.
authors.forename: Emma
authors.initials: E
authors.lastname: Raitoharju
authors.affiliation: Pathology, Fimlab Laboratories, Tampere University Hospital, Tampere, Finland.
authors.forename: Ivana
authors.initials: I
authors.lastname: Kholova
authors.affiliation: Heart Center, Tampere University Hospital, Tampere, Finland.
authors.forename: Thanos
authors.initials: T
authors.lastname: Sioris
authors.affiliation: School of Medicine, University of Tampere, Finland; Division of Vascular Surgery, Department of Surgery, Tampere University Hospital, Finland; Department of Clinical Physiology, Tampere University Hospital, Tampere, Finland.
authors.forename: Mika
authors.initials: M
authors.lastname: Kähönen
authors.affiliation: Department of Clinical Chemistry, Fimlab Laboratories, Tampere University Hospital, Tampere, Finland; School of Medicine, University of Tampere, Finland.
authors.forename: Terho
authors.initials: T
authors.lastname: Lehtimäki
authors.affiliation: BioMediTech, University of Tampere, Tampere, Finland and Fimlab Laboratories, Tampere University Hospital, Tampere, Finland.
authors.forename: Vesa P
authors.initials: VP
authors.lastname: Hytönen
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D001012:Aorta, Abdominal / Q000737:chemistry / Q000473:pathology; D001018:Aortic Diseases / Q000175:diagnosis / Q000235:genetics / Q000378:metabolism*; D050197:Atherosclerosis / Q000175:diagnosis / Q000235:genetics / Q000378:metabolism*; D015415:Biomarkers / Q000032:analysis; D018821:CD18 Antigens / Q000032:analysis* / Q000235:genetics; D002339:Carotid Arteries / Q000737:chemistry / Q000473:pathology; D002340:Carotid Artery Diseases / Q000175:diagnosis / Q000235:genetics / Q000378:metabolism*; D016022:Case-Control Studies; D016000:Cluster Analysis; D005260:Female; D005263:Femoral Artery / Q000737:chemistry / Q000473:pathology; D005455:Fluorescent Antibody Technique; D020869:Gene Expression Profiling / Q000379:methods; D006801:Humans; D007249:Inflammation / Q000175:diagnosis / Q000235:genetics / Q000378:metabolism*; D018836:Inflammation Mediators / Q000032:analysis; D008264:Macrophages / Q000737:chemistry* / Q000473:pathology; D008297:Male; D008565:Membrane Proteins / Q000032:analysis* / Q000235:genetics; D008875:Middle Aged; D009363:Neoplasm Proteins / Q000032:analysis* / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D010641:Phenotype; D058226:Plaque, Atherosclerotic; D060888:Real-Time Polymerase Chain Reaction; D012422:Rupture, Spontaneous; D015854:Up-Regulation
chemical_list: D015415:Biomarkers; D018821:CD18 Antigens; C477322:FERMT3 protein, human; D018836:Inflammation Mediators; D008565:Membrane Proteins; D009363:Neoplasm Proteins
keywords: Atherosclerosis; Gene expression; Immunohistochemistry; Integrin; Kindlin