Levels of extra domain A containing fibronectin in human atherosclerotic plaques are associated with a stable plaque phenotype



      Extra domain A (EDA), splice-variant of fibronectin, is a Toll-like receptor 4 (Tlr4) ligand. Recently, EDA has been demonstrated to enhance atherogenesis in mice but human data on the role of EDA in atherosclerotic disease are lacking. We hypothesized that EDA is associated with unstable plaque phenotypes and that plasma EDA could serve as biomarker for atherosclerosis.


      EDA levels were assessed in carotid endarterectomy specimen (206 patients) and related with plaque phenotype. In a second patient cohort, systemic EDA levels in atherosclerotic patients (73 patients) were compared to risk-factor matched controls (68 patients).


      EDA plaque levels were associated with characteristics of stable plaques; more smooth muscle cells (P = 0.003), more collagen (P = 0.071) and less fat (P = 0.023). Concomitantly, asymptomatic patients showed higher EDA values in the plaque compared to symptomatic patients (P = 0.004). EDA plasma levels did not differ between atherosclerotic patients versus controls (P = 0.134).


      EDA plaque levels are higher in asymptomatic patients and are associated with a stable plaque phenotype. EDA is not a plasma marker for atherosclerotic disease. These results suggest that local presence of endogenous Tlr4 ligand EDA is not associated with in an unstable plaque phenotype in humans.


      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'


      Subscribe to Atherosclerosis
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect


        • Hynes R.O.
        Fibronectins. Springer-Verlag, New York1990
        • Brown L.F.
        • Dubin D.
        • Lavigne L.
        • et al.
        Macrophages and fibroblasts express embryonic fibronectins during cutaneous wound healing.
        Am J Pathol. 1993; 142: 793-801
        • Ffrench-Constant C.
        • Van De W.L.
        • Dvorak H.F.
        • Hynes R.O.
        Reappearance of an embryonic pattern of fibronectin splicing during wound healing in the adult rat.
        J Cell Biol. 1989; 109: 903-914
        • Van Der Straaten H.M.
        • Canninga-van Dijk M.
        • Verdonck L.F.
        • et al.
        Extra-domain-A fibronectin: a new marker of fibrosis in cutaneous graft-versus-host disease.
        J Invest Dermatol. 2004; 123: 1057-1062
        • Barnes J.L.
        • Hastings R.R.
        • De la Garza M.A.
        Sequential expression of cellular fibronectin by platelets, macrophages, and mesangial cells in proliferative glomerulonephritis.
        Am J Pathol. 1994; 145: 585-597
        • Jarnagin W.R.
        • Rockey D.C.
        • Koteliansky V.E.
        • Wang S.S.
        • Bissell D.M.
        Expression of variant fibronectins in wound healing: cellular source and biological activity of the EIIIA segment in rat hepatic fibrogenesis.
        J Cell Biol. 1994; 127: 2037-2048
        • Kuhn C3
        • Boldt J.
        • King T.E.J.
        • et al.
        An immunohistochemical study of architectural remodeling and connective tissue synthesis in pulmonary fibrosis.
        Am Rev Respir Dis. 1989; 140: 1693-1703
        • Kanters S.D.
        • Banga J.D.
        • Algra A.
        • et al.
        Plasma levels of cellular fibronectin in diabetes.
        Diabetes Care. 2001; 24: 323-327
        • Peters J.H.
        • Carsons S.
        • Kalunian K.
        • et al.
        Preferential recognition of a fragment species of osteoarthritic synovial fluid fibronectin by antibodies to the alternatively spliced EIIIA segment.
        Arthritis Rheum. 2001; 44: 2572-2585
        • Coito A.J.
        • Brown L.F.
        • Peters J.H.
        • Kupiec-Weglinski J.W.
        • Van De Water L.
        Expression of fibronectin splicing variants in organ transplantation: a differential pattern between rat cardiac allografts and isografts.
        Am J Pathol. 1997; 150: 1757-1772
        • Dubin D.
        • Peters J.H.
        • Brown L.F.
        • et al.
        Balloon catheterization induced arterial expression of embryonic fibronectins.
        Arterioscler Thromb Vasc Biol. 1995; 15: 1958-1967
        • Glukhova M.A.
        • Frid M.G.
        • Shekhonin B.V.
        • et al.
        Expression of extra domain A fibronectin sequence in vascular smooth muscle cells is phenotype dependent.
        J Cell Biol. 1989; 109: 357-366
        • Takasaki I.
        • Chobanian A.V.
        • Mamuya W.S.
        • Brecher P.
        Hypertension induces alternatively spliced forms of fibronectin in rat aorta.
        Hypertension. 1992; 20: 20-25
        • Okamura Y.
        • Watari M.
        • Jerud E.S.
        • et al.
        The extra domain A of fibronectin activates Toll-like receptor 4.
        J Biol Chem. 2001; 276: 10229-10233
        • Poltorak A.
        • He X.
        • Smirnova I.
        • et al.
        Defective LPS signaling in C3H/HeJ and C57BL/10ScCr mice: mutations in Tlr4 gene.
        Science. 1998; 282: 2085-2088
        • Ohashi K.
        • Burkart V.
        • Flohe S.
        • Kolb H.
        Cutting edge: heat shock protein 60 is a putative endogenous ligand of the toll-like receptor-4 complex.
        J Immunol. 2000; 164: 558-561
        • Xu X.H.
        • Shah P.K.
        • Faure E.
        • et al.
        Toll-like receptor-4 is expressed by macrophages in murine and human lipid-rich atherosclerotic plaques and upregulated by oxidized LDL.
        Circulation. 2001; 104: 3103-3108
        • Hollestelle S.C.
        • De Vries M.R.
        • Van Keulen J.K.
        • et al.
        Toll-like receptor 4 is involved in outward arterial remodeling.
        Circulation. 2004; 109: 393-398
        • Michelsen K.S.
        • Wong M.H.
        • Shah P.K.
        • et al.
        Lack of Toll-like receptor 4 or myeloid differentiation factor 88 reduces atherosclerosis and alters plaque phenotype in mice deficient in apolipoprotein E.
        Proc Natl Acad Sci USA. 2004; 101: 10679-10684
        • Pasterkamp G.
        • Galis Z.S.
        • De Kleijn D.P.
        Expansive arterial remodeling: location, location, location.
        Arterioscler Thromb Vasc Biol. 2004; 24: 650-657
        • Vink A.
        • Schoneveld A.H.
        • Van Der Meer J.J.
        • et al.
        In vivo evidence for a role of toll-like receptor 4 in the development of intimal lesions.
        Circulation. 2002; 106: 1985-1990
        • Wright S.D.
        • Burton C.
        • Hernandez M.
        • et al.
        Infectious agents are not necessary for murine atherogenesis.
        J Exp Med. 2000; : 1437-1442
        • Tan M.H.
        • Sun Z.
        • Opitz S.L.
        • et al.
        Deletion of the alternatively spliced fibronectin EIIIA domain in mice reduces atherosclerosis.
        Blood. 2004; 1: 11-18
        • Verhoeven B.A.
        • Velema E.
        • Schoneveld A.H.
        • et al.
        Athero-express: differential atherosclerotic plaque expression of mRNA and protein in relation to cardiovascular events and patient characteristics. Rationale and design.
        Eur J Epidemiol. 2004; 19: 1127-1133
        • Simons P.C.
        • Algra A.
        • van de Laak M.F.
        • Grobbee D.E.
        • Van Der G.Y.
        Second manifestations of ARTerial disease (SMART) study: rationale and design.
        Eur J Epidemiol. 1999; 15: 773-781
        • Peters J.H.
        • Maunder R.J.
        • Woolf A.D.
        • Cochrane C.G.
        • Ginsberg M.H.
        Elevated plasma levels of ED1+ (“cellular”) fibronectin in patients with vascular injury.
        J Lab Clin Med. 1989; 113: 586-597
        • Sluijter J.P.
        • Pulskens W.P.
        • Schoneveld A.H.
        • et al.
        Matrix metalloproteinase 2 is associated with stable and matrix metalloproteinases 8 and 9 with vulnerable carotid atherosclerotic lesions: a study in human endarterectomy specimen pointing to a role for different extracellular matrix metalloproteinase inducer glycosylation forms.
        Stroke. 2006; 37: 235-239
        • Verhoeven B.
        • Hellings W.E.
        • Moll F.L.
        • et al.
        Carotid atherosclerotic plaques in patients with transient ischemic attacks and stroke have unstable characteristics compared with plaques in asymptomatic and amaurosis fugax patients.
        J Vasc Surg. 2005; 42: 1075-1081
        • Singh P.
        • Reimer C.L.
        • Peters J.H.
        • et al.
        The spatial and temporal expression patterns of integrin alpha9beta1 and one of its ligands, the EIIIA segment of fibronectin, in cutaneous wound healing.
        J Invest Dermatol. 2004; 123: 1176-1181
        • Sasu S.
        • LaVerda D.
        • Qureshi N.
        • Golenbock D.T.
        • Beasley D.
        Chlamydia pneumoniae and chlamydial heat shock protein 60 stimulate proliferation of human vascular smooth muscle cells via toll-like receptor 4 and p44/p42 mitogen-activated protein kinase activation.
        Circ Res. 2001; 89: 244-250
        • Edfeldt K.
        • Swedenborg J.
        • Hansson G.K.
        • Yan Z.Q.
        Expression of toll-like receptors in human atherosclerotic lesions: a possible pathway for plaque activation.
        Circulation. 2002; 105: 1158-1161
        • Strauss B.H.
        • Chisholm R.J.
        • Keeley F.W.
        • et al.
        Extracellular matrix remodeling after balloon angioplasty injury in a rabbit model of restenosis.
        Circ Res. 1994; 75: 650-658