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Prepublished online as a Blood First Edition Paper on July 3, 2003; DOI 10.1182/blood-2003-01-0306.
Submitted January 30, 2003
Accepted June 4, 2003
Vasculin, a novel vascular protein differentially expressed in human atherogenesis
Ann P J J Bijnens, Ann Gils, Barry Jutten, Birgit C G Faber, Sylvia Heeneman, Peter J E H M Kitslaar, Jan H M Tordoir, Carlie J M de Vries, Abraham A Kroon, Mat J A P Daemen*, and Kitty B J M Cleutjens
Department of Pathology, Cardiovascular Research Institute Maastricht (CARIM), University of Maastricht, Maastricht, The Netherlands
Department of General Surgery, Cardiovascular Research Institute Maastricht (CARIM), University of Maastricht, Maastricht, The Netherlands
Department of Internal Medicine, Cardiovascular Research Institute Maastricht (CARIM), University of Maastricht, Maastricht, The Netherlands
Department of Biochemistry, Academical Medical Center, University of Amsterdam, Amsterdam, The Netherlands
* Corresponding author; email: Mat_Daemen{at}yahoo.com.
Recent Suppressive Subtractive Hybridization analysis on human atherosclerotic plaque-derived RNA, revealed genes upregulated in plaques with a thrombus versus stable plaques. Clone SSH6, containing part of a putative open reading frame of an unknown protein, was further investigated. Full-length cDNA, coding for a 473 AA protein, was identified in a vascular Smooth Muscle Cell (SMC) cDNA library. Bio-informatics suggested the presence of multiple SSH6 variants due to alternative splicing of exon 3. Multiple tissue Northern blot analysis demonstrated a differential expression pattern of these variants, as a ubiquitously expressed SSH6 mRNA, missing exon 3, was detected apart from a putative vascular SMC-specific form containing exon 3. Western blot analysis indicated a ubiquitous 35 kDa protein (SSH6- ), besides a 45 kDa protein (vasculin), detected in the vascular wall and in plasma. Analysis of arteries displaying various stages of atherosclerosis, indicated that the ratio vasculin/SSH6- increases throughout atherogenesis. Immunohistochemical analysis demonstrated cytoplasmatic expression of SSH6 gene products in macrophages, endothelial and SMC. In summary, we identified a novel mRNA/protein, vasculin, in the arterial wall and plasma. The regulated expression of vasculin in plaques suggests a role in atherogenesis. Moreover, its presence in plasma opens perspectives for vasculin as a marker for atherosclerosis.
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