title: Target sequencing, cell experiments, and a population study establish endothelial nitric oxide synthase (eNOS) gene as hypertension susceptibility gene.
abstract: 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.
authors.affiliation: Studies Coordinating Centre, Research Unit Hypertension and Cardiovascular Epidemiology, KU Leuven Department of Cardiovascular Sciences, Campus Sint Rafaël, Kapucijnenvoer 35, Block D, Box 7001, BE-3000 Leuven, Belgium. jan.staessen@med.kuleuven.be.
authors.forename: Erika
authors.initials: E
authors.lastname: Salvi
authors.forename: Tatiana
authors.initials: T
authors.lastname: Kuznetsova
authors.forename: Lutgarde
authors.initials: L
authors.lastname: Thijs
authors.forename: Sara
authors.initials: S
authors.lastname: Lupoli
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Stolarz-Skrzypek
authors.forename: Francesca
authors.initials: F
authors.lastname: D'Avila
authors.forename: Valerie
authors.initials: V
authors.lastname: Tikhonoff
authors.forename: Silvia
authors.initials: S
authors.lastname: De Astis
authors.forename: Matteo
authors.initials: M
authors.lastname: Barcella
authors.forename: Jitka
authors.initials: J
authors.lastname: Seidlerová
authors.forename: Paola
authors.initials: P
authors.lastname: Benaglio
authors.forename: Sofia
authors.initials: S
authors.lastname: Malyutina
authors.forename: Francesca
authors.initials: F
authors.lastname: Frau
authors.forename: Dinesh
authors.initials: D
authors.lastname: Velayutham
authors.forename: Roberta
authors.initials: R
authors.lastname: Benfante
authors.forename: Laura
authors.initials: L
authors.lastname: Zagato
authors.forename: Alexandra
authors.initials: A
authors.lastname: Title
authors.forename: Daniele
authors.initials: D
authors.lastname: Braga
authors.forename: Diana
authors.initials: D
authors.lastname: Marek
authors.forename: Kalina
authors.initials: K
authors.lastname: Kawecka-Jaszcz
authors.forename: Edoardo
authors.initials: E
authors.lastname: Casiglia
authors.forename: Jan
authors.initials: J
authors.lastname: Filipovsky
authors.forename: Yuri
authors.initials: Y
authors.lastname: Nikitin
authors.forename: Carlo
authors.initials: C
authors.lastname: Rivolta
authors.forename: Paolo
authors.initials: P
authors.lastname: Manunta
authors.forename: Jacques S
authors.initials: JS
authors.lastname: Beckmann
authors.forename: Cristina
authors.initials: C
authors.lastname: Barlassina
authors.forename: Daniele
authors.initials: D
authors.lastname: Cusi
authors.forename: Jan A
authors.initials: JA
authors.lastname: Staessen
mesh_terms: D000328:Adult; D000483:Alleles; D001794:Blood Pressure / Q000235:genetics*; D016022:Case-Control Studies; D004730:Endothelium, Vascular / Q000503:physiopathology; D005260:Female; D020022:Genetic Predisposition to Disease; D005838:Genotype; D006801:Humans; D006973:Hypertension / Q000235:genetics* / Q000503:physiopathology; D008297:Male; D008875:Middle Aged; D052250:Nitric Oxide Synthase Type III / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide; D011401:Promoter Regions, Genetic; D044465:White People / Q000235:genetics
chemical_list: D052250:Nitric Oxide Synthase Type III
keywords: blood pressure; endothelial nitric oxide synthase gene; hypertension; population science; target sequencing; transfection