Atherosclerosis 206 (2009) 209–215 Contents lists available at ScienceDirect Atherosclerosis journal homepage: www.elsevier.com/locate/atherosclerosis Osteopontin gene variation and cardio/cerebrovascular disease phenotypes Klaus Schmidt-Petersen a , Eva Brand b , Ralph Telgmann a , Viviane Nicaud c , Claudia Hagedorn a , Julien Labreuche j , Corinna Dördelmann a , Alexis Elbaz d , Marion Gautier-Bertrand d , Jens W. Fischer e , Alun Evans f , Caroline Morrison g , Dominique Arveiler h , Monika Stoll i , Pierre Amarenco j , Franc ¸ ois Cambien c , Martin Paul k , Stefan-Martin Brand-Herrmann a,∗ a Leibniz-Institute for Arteriosclerosis Research, Department of Molecular Genetics of Cardiovascular Disease, University of Muenster, Germany b Department of Internal Medicine D, University Clinic of Muenster, Germany c INSERM, UMR S 525, Université Pierre et Marie Curie-Paris 6, UMR S 525, Paris 75634, France d Inserm, U708, Neuroepidemiology, Hôpital de la Salpêtrière, Université Pierre et Marie Curie-Paris 6, Paris 75013, France e Institut für Pharmakologie, Universitätsklinikum Essen Universität Duisburg-Essen, Germany f MONICA Project in Belfast, UK g MONICA Project in Glasgow, UK h MONICA Project in Strasbourg, France i Leibniz-Institute for Arteriosclerosis Research, Genetic Epidemiology of Vascular Disorders, University of Muenster, Germany j Department of Neurology and Stroke Centre, Bichat University Hospital, Paris, France k Maastricht University, Faculty of Health, Medicine, and Life Science, Maastricht, The Netherlands article info Article history: Received 24 October 2008 Received in revised form 22 January 2009 Accepted 9 February 2009 Available online 21 February 2009 Keywords: Osteopontin OPN variation Cardio and cerebrovascular disease Brain infarction Myocardial infarction abstract We aimed at associating common osteopontin (OPN) gene variants with cardiovascular disease phe- notypes.We scanned the OPN gene in 190 chromosomes from myocardial infarction (MI) patients and identified five variants in the promoter, three synonymous and one non-synonymous variant. All variants were investigated in case–control studies for MI (ECTIM: 990 cases, 900 controls) and brain infarction (BI) (GÉNIC: 466 cases, 444 controls). Promoter variants were functionally analyzed by bandshift assays, the coding D147D [T/C] by Western blot. Allele D147D C was independently and significantly associated with lower apoB levels (P = 0.044 [ECTIM] P =0.03 [GENIC]), its allele frequency was significantly lower in patients with BI compared to controls (OR [95% CI] 0.39 [0.20–0.74], P = 0.004), and C allele carriers had a significantly lower frequency of presence of carotid plaques (P = 0.02). Bandshifts with HepG2 and Ea.hy926 nuclear proteins did not reveal any functionality of promoter variants, whereas the OPN-441C- containing construct resulted in reduced OPN protein expression in Western blots, complying with its potential protective effect on the phenotypes studied.We here provide evidence that a portion of the OPN locus is likely to associate with cardiovascular disease-related phenotypes. However, further experiments are warranted to clarify the functional role of OPN variants. © 2009 Elsevier Ireland Ltd. All rights reserved. 1. Introduction Osteopontin (OPN, OMIM *166490) is a phosphorylated matrix glycoprotein that contains an Arg–Gly–Asp (RGD) cell attach- ment sequence [1] and promotes adhesion and migration of vascular smooth muscle cells (VSMCs) and endothelial cells in vitro [2]. With respect to the cardiovascular system and via dis- ∗ Corresponding author at: Leibniz-Institute for Arteriosclerosis Research, Uni- versity of Muenster, Department of Molecular Genetics of Cardiovascular Disease, Domagkstraße 3, D-48149 Muenster, Germany. Tel.: +49 251 83 52996; fax: +49 251 83 52997. E-mail address: brandher@uni-muenster.de (S.-M. Brand-Herrmann). tinct receptors [3], OPN participates in SMC, endothelial cell and leukocyte adhesion/migration within vascular injury sites [4], ele- vated OPN mRNA and protein expression has been demonstrated in human aortic, coronary and carotic atherosclerotic plaques [5], mostly accompanied with significant calcification. As SMCs and endothelial cells express OPN at very low levels, the major source of OPN in atherosclerotic plaques was reported to be infil- trating macrophages [6]. Transgenic mice overexpressing OPN display medial thickening and neointimal formation [7], and OPN- deficiency attenuates atherosclerosis, but associates with vascular calcification [8]. The human OPN gene is located on chromosome 4q21–q25, spans about 11kb, and consists of seven exons; the protein is composed of 314 amino acid residues, and splice variants have been described [9]. The aim of the present study was to 0021-9150/$ – see front matter © 2009 Elsevier Ireland Ltd. All rights reserved. doi:10.1016/j.atherosclerosis.2009.02.015