Target Sequencing, Cell Experiments, and a Population Study Establish Endothelial Nitric Oxide Synthase (eNOS) Gene as Hypertension Susceptibility Gene

• Published in: Hypertension (Dallas, Tex. 1979) Vol. 62; no. 5; pp. 844 - 852
• Main Authors: Salvi, Erika, Kuznetsova, Tatiana, Thijs, Lutgarde, Lupoli, Sara, Stolarz-Skrzypek, Katarzyna, D’Avila, Francesca, Tikhonoff, Valerie, De Astis, Silvia, Barcella, Matteo, Seidlerová, Jitka, Benaglio, Paola, Malyutina, Sofia, Frau, Francesca, Velayutham, Dinesh, Benfante, Roberta, Zagato, Laura, Title, Alexandra, Braga, Daniele, Marek, Diana, Kawecka-Jaszcz, Kalina, Casiglia, Edoardo, Filipovský, Jan, Nikitin, Yuri, Rivolta, Carlo, Manunta, Paolo, Beckmann, Jacques S, Barlassina, Cristina, Cusi, Daniele, Staessen, Jan A
• Format: Journal Article
• Language: English
• Published: Hagerstown, MD American Heart Association, Inc 01.11.2013; Lippincott Williams & Wilkins
• Subjects: Adult; Alleles; Arterial hypertension. Arterial hypotension; Biological and medical sciences; Blood and lymphatic vessels; Blood Pressure - genetics; Cardiology. Vascular system; Case-Control Studies; Endothelium, Vascular - physiopathology; European Continental Ancestry Group - genetics; Female; Genetic Predisposition to Disease; Genotype; Humans; Hypertension - genetics; Hypertension - physiopathology; Male; Medical sciences; Middle Aged; Nitric Oxide Synthase Type III - genetics; Polymorphism, Single Nucleotide; Promoter Regions, Genetic; Cardiovascular disease; target sequencing; Genetic determinism; Target; Transfection; Gene; Sciences; Predisposition; Genetics; Arterial pressure; endothelial nitric oxide synthase gene; Blood pressure; Target cell; Cardiology; Sequencing; population science; Human; Hypertension; Nucleotide sequence; Enzyme; Nitric-oxide synthase; Endothelium; Treatment; Cell population; Hemodynamics; Oxidoreductases; Circulatory system; Gene therapy; blood pressure; hypertension; transfection
• ISSN: 0194-911X; 1524-4563
• DOI: 10.1161/HYPERTENSIONAHA.113.01428

A case–control study revealed association between hypertension and rs3918226 in the endothelial nitric oxide synthase (eNOS) gene promoter (minor/major allele, T/C allele). We aimed at substantiating these preliminary findings by target sequencing, cell experiments, and a population study. We sequenced the 140-kb genomic area encompassing the eNOS gene. In HeLa and HEK293T cells transfected with the eNOS promoter carrying either the T or the C allele, we quantified transcription by luciferase assay. In 2722 randomly recruited Europeans (53.0% women; mean age 40.1 years), we studied blood pressure change and incidence of hypertension in relation to rs3918226, using multivariable-adjusted models. Sequencing confirmed rs3918226, a binding site of E-twenty six transcription factors, as the single nucleotide polymorphism most closely associated with hypertension. In T compared with C transfected cells, eNOS promoter activity was from 20% to 40% (P<0.01) lower. In the population, systolic/diastolic blood pressure increased over 7.6 years (median) by 9.7/6.8 mm Hg in 28 TT homozygotes and by 3.8/1.9 mm Hg in 2694 C allele carriers (P≤0.0004). The blood pressure rise was 5.9 mm Hg systolic (confidence interval [CI], 0.6–11.1; P=0.028) and 4.8 mm Hg diastolic (CI, 1.5–8.2; P=0.0046) greater in TT homozygotes, with no differences between the CT and CC genotypes (P≥0.90). Among 2013 participants normotensive at baseline, 692 (34.4%) developed hypertension. The hazard ratio and attributable risk associated with TT homozygosity were 2.04 (CI, 1.24–3.37; P=0.0054) and 51.0%, respectively. In conclusion, rs3918226 in the eNOS promoter tags a hypertension susceptibility locus, TT homozygosity being associated with lesser transcription and higher risk of hypertension.