The glu298asp polymorphism in the nitric oxide synthase 3 gene is associated with the risk of ischemic stroke in two large independent case–control studies
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The search for genes involved in the pathogenesis of stroke has been highlighted as a field of needs. We followed the concept, that stroke represents a complex genetic disorder, and analyzed the contribution of 106 informative single nucleotide polymorphisms (SNPs) from 63 candidate genes for cardiovascular diseases for the risk of stroke. We conducted two independent case–control studies in two different German regions and recruited a total of 1,901 hospitalized stroke cases and 1,747 regional population controls. The smaller of both studies was used as the replication study. Multiplex PCR in combination with allele-specific hybridization was used for genotype determination. Descriptive statistics, permutations and multivariable logistic regression were used in the analyses. After permutation testing 5 SNPs, located in the nitric oxide synthase 3, the alpha 2 integrin, the interleukin 13, the selectin P and the chemokine receptor 2 genes, had a significant allele difference between cases and controls in the larger study. For one of these SNPs, the glu298asp polymorphism in the nitric oxide synthase 3 gene, an association with ischemic stroke was replicated in the second study and also in a combined analysis of both studies. This association was independent of age, gender, hypertension, diabetes and hypercholesterolemia in both studies. Using large sample sizes and a replication study approach, we found evidence for a role of a polymorphism in the nitric oxide synthase 3 gene in stroke onset.
KeywordsIschemic Stroke Stroke Risk Replication Study Cerebral Small Vessel Disease Glu298asp Polymorphism
The contributions of the following hospitals (Departments of Neurology) are greatfully acknowledged (in alphabetical order): Bathildis Krankenhaus Bad Pyrmont, Universitätsklinikum Bochum im St. Josef-Hospital, Evangelisches Krankenhaus Castrop-Rauxel, St. Elisabeth-Stift Damme, Klinikum Dortmund, Hans-Susemihl-Krankenhaus Emden, Evangelisches Krankenhaus Gelsenkirchen, Universitätsklinikum Greifswald, St. Johannes Hospital Hagen, Marienhospital Hamm, Friederikenstift Hannover, Klinikum Minden, Universitätsklinikum Münster, Klinikum Neubrandenburg, Klinikum Osnabrück, St. Vincenz-Krankenhaus Paderborn, Prosperhospital Recklinghausen (Dept. of Geriatric Medicine), Ammerlandklinik Westerstede. We thank Elisabeth Lange for laboratory assistance, Gregor Kuhlenbäumer for methodological advice and Gabriele Beer, Michael Grow, Arkadiy Silbergleit, Rebecca Reynolds, Lori Steiner and Karen Walker for their expertise in developing the multiplex genotyping assays. This project is part of the German ‘Competence Net Stroke,’ which is supported by the German Federal Ministry of Education and Research (01GI9909/3). The Study of Health in Pomerania (SHIP) is funded by grants from the German Federal Ministry of Education and Research (BMBF, 01ZZ96030), and from the Ministry for Education, Research and Cultural Affairs and the Ministry for Social Affairs of the Federal State of Mecklenburg-Vorpommern. All multi-locus genotyping assays were provided by Roche Molecular Systems, Alameda, CA, USA.
- Chen W, Srinivasan SR, Elkasabany A, Ellsworth DL, Boerwinkle E, Berenson GS (2001) Combined effects of endothelial nitric oxide synthase gene polymorphism (G894T) and insulin resistance status on blood pressure and familial risk of hypertension in young adults: the Bogalusa Heart Study. Am J Hypertens 14:1046–1052PubMedCrossRefGoogle Scholar
- Dahlman I, Eaves IA, Kosoy R, Morrison VA, Heward J, Gough SC, Allahabadia A, Franklyn JA, Tuomilehto J, Tuomilehto-Wolf E, Cucca F, Guja C, Ionescu-Tirgoviste C, Stevens H, Carr P, Nutland S, McKinney P, Shield JP, Wang W, Cordell HJ, Walker N, Todd JA, Concannon P (2002) Parameters for reliable results in genetic association studies in common disease. Nat Genet 30:149–150PubMedCrossRefGoogle Scholar
- Fisher (1932) Statistical methods for research workers. Oliver & Boyd, EdingburghGoogle Scholar
- Forstermann U, Pollock JS, Schmidt HH, Heller M, Murad F (1991) Calmodulin-dependent endothelium-derived relaxing factor/nitric oxide synthase activity is present in the particulate and cytosolic fractions of bovine aortic endothelial cells. Proc Natl Acad Sci USA 88:1788–1792PubMedCrossRefGoogle Scholar
- Henskens LH, Kroon AA, van Boxtel MP, Hofman PA, de Leeuw PW (2005) Associations of the angiotensin II type 1 receptor A1166C and the endothelial NO synthase G894T gene polymorphisms with silent subcortical white matter lesions in essential hypertension. Stroke 36:1869–1873PubMedCrossRefGoogle Scholar
- Higuchi (1989) PCR technology, principles and applications for DNA amplification. Stockton Press, New York, pp 31–38Google Scholar
- Howard TD, Giles WH, Xu J, Wozniak MA, Malarcher AM, Lange LA, Macko RF, Basehore MJ, Meyers DA, Cole JW, Kittner SJ (2005) Promoter polymorphisms in the nitric oxide synthase 3 gene are associated with ischemic stroke susceptibility in young black women. Stroke 36:1848–1851PubMedCrossRefGoogle Scholar
- John U, Greiner B, Hensel E, Ludemann J, Piek M, Sauer S, Adam C, Born G, Alte D, Greiser E, Haertel U, Hense HW, Haerting J, Willich SN, Kessler C (2001) Study of health in Pomerania (SHIP): a health examination survey in an east German region: objectives and design. Soz Präventivmed 46:186–194PubMedCrossRefGoogle Scholar
- Tesauro M, Thompson WC, Rogliani P, Qi L, Chaudhary PP, Moss J (2000) Intracellular processing of endothelial nitric oxide synthase isoforms associated with differences in severity of cardiopulmonary diseases: cleavage of proteins with aspartate vs. glutamate at position 298. Proc Natl Acad Sci USA 97:2832–2835PubMedCrossRefGoogle Scholar