eNOS Single Nucleotide Polymorphisms Research Articles

Interaction between eNOS gene polymorphism and current smoking on susceptibility to coronary heart disease in Chinese people.

This study aims to explore the relation between endothelial nitric oxide synthase (eNOS) single-nucleotide polymorphisms (SNPs) and the risk of coronary heart disease (CHD). SNPstats (online software: http://bioinfo.iconcologia.net/SNPstats) was performed to test Hardy-Weinberg equilibrium in controls. Generalized multifactor dimensionality reduction (GMDR) was adopted to screen the preferable interaction between eNOS SNPs and smoking. The frequency for the rs1799983-T allele was 31.1% in CHD patients, which was significantly higher than that of 19.8% in controls (P < 0.05). The frequency for the rs891512-A allele was 28.8% in cases, which was also significantly higher than that of 20.1% in controls (P < 0.05). Logistic regression analysis showed that both rs1799983-T and rs891512-A alleles were related with increased risk of CHD, and the odds ratios (ORs) [95% confidence interval (CI)] were 1.71 (1.31-2.15) and 1.57 (1.14-2.07), respectively. High-order interactions were investigated among SNPs and environmental factors using the GMDR method. The data showed that a two-locus model (rs1799983 × smoking) had a testing accuracy of 0.60 (P = 0.001). We found that current smokers with rs1799983-GT or TT within eNOS gene have the highest CHD risk, compared to never smokers with rs1799983-GG genotype, OR (95% CI) = 2.74 (1.78-3.85), after covariates adjustment for age, gender, BMI, and alcohol drinking. The rs1799983-T and rs891512-A alleles and interaction between rs1799983 and smoking were all risk factors of CHD.

Impact of endothelial nitric oxide synthase polymorphisms on urothelial cell carcinoma development

ObjectivesUrothelial cell carcinoma (UCC), a major malignancy of the genitourinary tract, is induced through carcinogenic etiological factors. Endothelial nitric oxide synthase (eNOS) is one of the major isoforms of nitric oxide synthase and is involved in various pathophysiologic and physiologic processes. In this study, eNOS single-nucleotide polymorphisms were investigated to evaluate UCC susceptibility and clinicopathological characteristics. Materials and methodsTwo single-nucleotide polymorphisms of eNOS in 431 patients with UCC and 862 controls without cancer were analyzed using real-time polymerase chain reaction. ResultsThe results showed that 272 men with UCC having eNOS 894 G > T rs1799983 “GT + TT” variants had a high risk of developing a large tumor (T1–T4, P = 0.038). Furthermore, a correlation was observed between the expressions of eNOS and invasive tumor, metastasis and poor survival in urothelial carcinoma in The Cancer Genome Atlas data set. ConclusionOur results indicated that male patients with UCC carrying eNOS 894 G > T rs1799983 “GT + TT” genetic variants have a high risk of developing a large tumor, and eNOS polymorphisms may serve as a marker or therapeutic target in UCC treatment.

Involvement of endothelial nitric oxide synthase gene variants in the aggressiveness of uterine cervical cancer

To date, few studies explore the involvement of endothelial nitric oxide synthase (eNOS) gene variants in uterine cervical cancer. Therefore, we conducted this study to assess the clinical implication of eNOS in cervical carcinogenesis, clinicopathological characteristics and patient survival. One hundred and seventeen patients with cervical invasive cancer and 95 with preinvasive lesions and 330 control women were consecutively enrolled. Real time polymerase chain reaction was used to examine the genotypic distributions of eNOS single nucleotide polymorphisms (SNPs) rs1799983 (894G>T) at the exon 7 region and rs2070744 (-786T>C) at the promoter region. Our results indicated no significant associations among genotypic distributions of eNOS SNPs and patients with cervical invasive cancer and those with preinvasive lesions as well as normal controls. However, cervical cancer patients with genotypes TC/CC in eNOS SNP rs2070744 carried less risk of advanced stage [odds ratios (OR) = 0.30, 95% confidence interval (CI)=0.09-0.97, p=0.036], parametrium invasion (OR=0.16, 95% CI=0.02-0.75, p=0.009) and pelvic lymph node metastasis (OR=0.12, 95% CI=0.01-0.89, p=0.016). In conclusion, although eNOS SNPs rs2070744 and rs1799983 do not display significant associations with cervical carcinogenesis and patient survival, cervical cancer patients with genotypes TC/CC in rs2070744 carry less risk of advanced stage, parametrium invasion and pelvic lymph node metastasis in Taiwan.

Associations of genetic variations of the endothelial nitric oxide synthase gene and environmental carcinogens with oral cancer susceptibility and development

Oral cancer is a major head and neck cancer that is reported to be causally associated with genetic factors and environmental carcinogens. Endothelial nitric oxide synthase (eNOS) was reported to modulate carcinogenesis and progression through nitric oxide (NO) production. Genetic polymorphisms in the eNOS gene can regulate its transcription and further mediate NO production. The purpose of this study was to explore the influences of eNOS gene polymorphisms combined with environmental carcinogens on the predisposition for oral cancer. Two single-nucleotide polymorphisms (SNPs) of the eNOS gene, −786 T > C (rs2070744) and 894G > T (rs1799983), were genotyped in 1200 controls and 1044 patients with oral cancer using a TaqMan-based real-time polymerase chain reaction (PCR). We found that patients who carried the −786 T > C TC genotype were at higher risk for developing an advanced clinical stage (stage III/IV) compared to those with the −786 T > C TT genotype; however, there was no significant association of the two individual SNPs with oral cancer between patients and the control group. According to behavioral exposure to environmental carcinogens, the presence of these two eNOS SNPs combined with tobacco use and/or betel quid chewing profoundly enhanced the risk of oral cancer. Moreover, carriers with the betel quid-chewing habit who had haplotypes of the two eNOS SNPs more easily developed oral cancer. These results indicated an involvement of −786 T > C polymorphisms in the progression of oral cancer and support the interaction between eNOS gene polymorphisms and environmental carcinogens as a predisposing factor of oral carcinogenesis.

Endothelial nitric oxide synthase gene single nucleotide polymorphisms and the risk of hypertension: A meta-analysis involving 63,258 subjects

ABSTRACTThe endothelial nitric oxide synthase (eNOS) gene plays an important role in regulating vascular tone and blood pressure. Recently, the eNOS G894T and T-786C single nucleotide polymorphisms (SNPs) were intensively studied with regard to their associations with hypertension. However, the results of these studies were inconsistent. Therefore, we conducted the so far largest meta-analysis to better assess the correlations between eNOS SNPs and hypertension. Eligible articles were searched in PubMed, Medline, Embase, Scopus, and CNKI up to April 2016. Odds ratios (ORs) and 95% confidence intervals (CIs) were used to detect any potential associations between eNOS SNPs and the risk of hypertension. A total of 95 case–control studies involving 29,308 hypertension cases and 33,950 healthy controls were analyzed. The overall meta-analysis results showed that eNOS G894T and T-786C SNPs were both significantly associated with the risk of hypertension, the T allele of G894T SNP (G versus T, P < 0.00001, OR = 0.82, 95% CI 0.76–0.89) and C allele of T-786C SNP (T versus C, P = 0.004, OR = 0.92, 95% CI 0.87–0.97) conferred an increased susceptibility to hypertension. Further subgroup analyses yielded similar positive results for G894T SNP in essential hypertension, gestational hypertension, and Asian ethnicity, and that for T-786C SNP in essential hypertension and Asian population. Overall, our findings suggest that eNOS G894T and T-786C SNPs were both significantly correlated with hypertension. Additionally, the T allele of G894T SNP and C allele of T-786C SNP may serve as potential biological markers for hypertension susceptibility in Asians.

The Endothelial Nitric Oxide Synthase Gene Polymorphism is Associated with the Susceptibility to Immunoglobulin a Nephropathy in Chinese Population

Background/Aims: Endothelial nitric oxide synthase (eNOS) is one of the most important enzymes for producting nitric oxide (NO), which regulate the function of many organs and cells. The single nucleotide polymorphisms (SNPs) of eNOS were found to be associated with many kidney diseases. However, it is lack of relevant studies to evaluate the associations between eNOS polymorphisms and immunoglobulin A nephropathy (IgAN). This case-control study aimed to evaluate the relationship between eNOS polymorphisms and IgAN. Methods: We recruited 351 IgAN patients and 310 age- and sex-matched healthy controls from Northwest China. Sequenom MassARRAY was used to detect the genotypes of two common eNOS SNPs (rs1799983 and rs2070744). Odds ratios (ORs) and 95% confidence intervals (95% CIs) were calculated by the Chi square test to evaluate the associations between eNOS and IgAN. Phase 2.1 was used to conduct haplotype analysis. Results: In the overall analysis, we found that the rs1799983 polymorphism was associated with a decreased risk of IgAN (G/T vs. G/G: OR=0.57, 95%CI=0.34–0.96; G/T+T/T vs. G/G: OR=0.52, 95%CI=0.31–0.86; G/T vs. G/G-T/T: OR=0.60, 95%CI=0.36–0.99; Log-additive model: OR=0.48, 95%CI=0.30–0.78). Haplotype analysis indicated that T_rs1799983C_rs2070744 is a protective factor against IgAN (OR=0.62, 95%CI=0.42––0.92). However, no significant differences were found between the two SNPs (rs1799983 and rs2070744) and clinical features (age, sex, blood pressure, and Lee’s grade) of IgAN. Conclusion: The eNOS gene rs1799983 polymorphism and T_rs1799983C_rs2070744 haplotype may reduce the risk of IgAN in Chinese populations.

Association of eNOS Polymorphisms with Anterior Chamber Depth in Han Chinese: Jiangsu Eye Study

Recently, a study reported that single nucleotide polymorphisms (SNP) in endothelial nitric oxide synthase (eNOS) were associated with primary angle closure glaucoma (PACG) in Australian cohort. In this study, we aimed to investigate whether those eNOS SNPs are associated with primary angle closure (PAC) or ocular biometric characteristics such as axial length (AL), anterior chamber depth (ACD), and diopter of spherical power (DS) in Han Chinese. The samples consisted of 232 PAC subjects and 306 controls collected from a population-based prevalence survey conducted in Funing County of Jiangsu, China. The rs3793342 and rs11771443 in eNOS were genotyped by TaqMan-MGB probe using the RT-PCR system. Our data did not identify any association of the eNOS SNPs with PAC. However, the analysis on the quantitative traits of ocular biometrics showed that the ACD of rs11771443 AA and GA carriers is significantly deeper than that of rs11771443 GG carriers (P = 0.0025), even though the AL and DS are not associated with rs11771443 genotypes. Rs3793342 was not associated with any biometric parameters including ACD, AL and DS. In summary, our data indicates that eNOS rs11771443 is associated with ACD and its role in the pathogenesis of PACG warranted further study.

A susceptibility haplotype within the endothelial nitric oxide synthase gene influences bone mineral density in hypertensive women

The influence of the coordinated effect of various single-nucleotide polymorphisms (SNPs) within the endothelial nitric oxide synthase (eNOS) gene on the risk of osteoporosis in hypertension has remained undetermined. Four pertinent SNPs of the eNOS gene, rs2070774, rs1799983, rs1800780 and rs3918181, were examined for the risk of osteoporosis in 313 hypertensive postmenopausal women in Northwest India. All the hypertensive women were verified with dual energy X-ray absorptiometry and categorized as 150 with osteoporosis and 163 without osteoporosis. The minor allele (T) of rs1799983 exerts a statistically significant risk for osteoporosis both in dominant [odds ratio (OR) 3.71, 95 % confidence interval (CI) 2.12-6.49, P < 0.001] and recessive mode (OR 5.75, 95 % CI 1.24-26.69, P = 0.036) after Bonferroni correction. Bone mineral density (BMD) values (corrected for the effects of risk variables) according to eNOS SNP genotypes revealed a significant association with rs1799983 at both the lumbar spine (P = 0.001) and femoral neck (P = 0.023). Risk association analyses revealed a susceptibility haplotype TTAG which influences the risk of osteoporosis (OR 2.02, 95 % CI 1.05-3.39, P = 0.042) in hypertension after adjusting for the effects of risk factors. Furthermore, this haplotype was significantly associated with BMD at the lumbar spine (P = 0.029) and femoral neck (P = 0.021) in a dose-dependent manner. The results suggest that possession of the TTAG haplotype of the eNOS gene may increase the risk of osteoporosis two-fold in hypertensive postmenopausal women in Northwest India.

Endothelial nitric oxide synthase gene polymorphisms and renal responsiveness to RAS inhibition therapy in type 2 diabetic Asian Indians

AimTo investigate the association of functional single nucleotide polymorphisms (SNPs) of the endothelial nitric oxide synthase gene (eNOS) gene (T-786C, G894T) and one variable number tandem repeat polymorphism (aa 27VNTR bb) with reno-protective response to angiotensin converting enzyme inhibitors (ACEI) and angiotensin receptor blockers (ARB) therapy in North Indian type 2 diabetic mellitus (T2DM) subjects with cases having diabetic nephropathy (DN) and controls without DN. MethodWe genotyped three polymorphisms of eNOS (two SNPs: T-786C, G894T and one 27 VNTR) in T2DM patients with overt nephropathy (cases: n=320) and T2DM patients without overt nephropathy (controls: n=490), using validated PCR-RFLP assays. These 810 North Indian T2DM patients treated with ACEI or ARB after diagnosis were followed up for 3 years. Percent changes in eGFR, urinary albumin excretion (UAE), serum creatinine at the end of 3 years of treatment were taken as end points of renoprotective response. ResultWe observed that in normoalbuminuric patients, eNOS -786 CC genotype and haplotypes C-b-G and C-b-T were associated with lesser renoprotective response to ACEI. While, in macroalbuminurics, eNOS -786 CC genotype, haplotypes C-b-G and C-b-T and 27VNTR aa were associated with better renoprotective response to ACEI/ARB. ConclusionOur results showed that eNOS T-786C CC genotype and 27VNTR individually and in interaction with other eNOS SNPs modulate renoprotective efficacy of ACEI and ARB in T2DM patients, depending on the status of proteinuria.

Association study of ACE and eNOS single nucleotide polymorphisms with Henoch-Schönlein purpura nephritis

Previous studies have shown that insertion/deletion polymorphisms in the angiotensin-converting enzyme (ACE) gene and the endothelial nitric oxide synthase (eNOS) gene are associated with Henoch-Schönlein purpura nephritis (HSPN). However, further studies are required to prove the relationship between HSPN and ACE and eNOS single nucleotide polymorphisms (SNPs). We studied six ACE SNPs and two eNOS SNPs by genotyping HSPN patients. Statistical analyses indicate that four ACE SNPs and two eNOS SNPs are associated with HSPN susceptibility. A cumulative effect analysis suggested that an increased number of unfavourable genotypes may lead to an increased risk of HSPN. By comparing alleles, genotypes and haplotypes that are associated with lupus nephritis (LN) and HSPN, we found genetic heterogeneity between HSPN and LN.

Effects of angiotensin receptor blockers on endothelial nitric oxide release: the role of eNOS variants

• Angiotensin II receptor blockers improve endothelial cell-dependent vasodilation in patients with hypertension through suppression of angiotensin II type 1 receptors but may have additional and differential effects on endothelial nitric oxide synthase (eNOS) function. • The key finding from this study is that angiotensin II receptor blockers (ARBs) differentially enhanced nitric oxide (NO) release in a manner influenced by certain genetic variants of eNOS. This finding provides new insights into the effects of ARBs on endothelial cell-dependent vasodilation and eNOS function that are of high importance in vascular medicine and clinical pharmacology. AIM Angiotensin II receptor blockers (ARBs) improve endothelial cell (EC)-dependent vasodilation in patients with hypertension through suppression of angiotensin II type 1 receptors but may have additional and differential effects on endothelial nitric oxide (NO) synthase (eNOS) function. To investigate this question, we tested the effects of various ARBs on NO release in ECs from multiple donors, including those with eNOS genetic variants linked to higher cardiovascular risk. The effects of ARBs (losartan, olmesartan, telmisartan, valsartan), at 1 µm, on NO release were measured with nanosensors in human umbilical vein ECs obtained from 18 donors. NO release was stimulated with calcium ionophore (1 µm) and its maximal concentration was correlated with eNOS variants. The eNOS variants were determined by a single nucleotide polymorphism in the promoter region (T-786C) and in the exon 7 (G894T), linked to changes in NO metabolism. RESULTS All of the ARBs caused an increase in NO release as compared with untreated samples (P < 0.01, n= 4-5 in all eNOS variants). However, maximal NO production was differentially influenced by eNOS genotype. Olmesartan increased maximal NO release by 30%, which was significantly greater (P < 0.01, n= 4-5 in all eNOS variants) than increases observed with other ARBs. The ARBs differentially enhanced NO release in ECs in a manner influenced by eNOS single nucleotide polymorphisms. These findings provide new insights into the effects of ARBs on EC-dependent vasodilation and eNOS function.

67: Single nucleotide polymorphisms (SNPs) in endothelial nitric oxide synthase (eNOS) gene and unexplained stillbirth

nitric oxide synthase (eNOS) gene and unexplained stillbirth Francesca Ferrari, Fabio Facchinetti, Huaizhi Yin, George Saade, Monica Longo Obstetrics and Gynaecology, University of Modena and Reggio Emilia, Mother-Infant department, Modena, Italy, The University of Texas Medical Branch, Obstetrics & Gynecology, Galveston, TX OBJECTIVE: Altered utero-placental perfusion has been implicated in the pathogenesis of unexplained stillbirth (SB). We previously reported that genetic polymorphisms of VEGF affecting placental vasculogenesis are associated with unexplained SB. Nitric oxide (NO) plays a critical role in regulation of uterine perfusion, and eNOS is the rate limiting enzyme in the NO production cascade. Previous studies suggest that eNOS gene SNPs may be associated with preeclampsia and idiopathic recurrent pregnancy loss. Our objective was to evaluate the presence and distribution of eNOS gene SNPs between unexplained SB and controls in a well-characterized cohort. STUDY DESIGN: Placentas were obtained from 50 unexplained SB and 46 livebirth controls. Classification of “unexplained” stillbirth was by Wigglesworth and Aberdeen criteria and included both appropriate (AGA-SB) and small for gestational age (SGA-SB, 10th%) SB. Placental DNA was extracted (Qiagen) and evaluated for 4 eNOS SNPs (rs1007311 A/G, rs891512 A/G, rs1800779 A/G and rs1800783 A/T) by real time PCR using specific taqman probes. The genotype and allelic distributions were compared between the groups using Chisquare (significance: p 0.05). RESULTS: All SNPs were in Hardy-Weinberg equilibrium. None of the SNPs were associated with SB overall. However, significant different genotype distribution emerged for eNOS-SNP rs1800783 A/T when comparing the subgroups of SGA-SB and AGA-SB to controls (p 0.004). Allele-A carriers of rs1800783 were more frequent in AGA-SB compared to both controls (p 0.003) and SGA-SB (p 0.001)(Fig. 1). CONCLUSION: This is the first study to report on placental eNOS polymorphisms and unexplained SB. Importantly we found that Allele A of rs1800783 eNOS SNP might play a role in the occurrence of unexplained stillbirth in the setting of adequate growth. Our results warrant further population-based analysis. 68 The influence of a mediolateral episiotomy during an operative vaginal delivery on the risk for Obstetric Anal Sphincter Injuries (OASIS) Joey de Vogel, Anneke van Beek, Dirk Gietelink, Marijana Vujkovic, Jan Willem de Leeuw, Dimitri Papatsonis Amphia hospital, Department of Obstetrics and Gynecology, Breda, Netherlands, Erasmus Medical Center, Department of Obstetrics and Gynecology, Rotterdam, Netherlands, Ikazia Hospital, Department of Obstetrics and Gynecology, Rotterdam, Netherlands OBJECTIVE: Operative vaginal delivery is associated with an increased risk of Obstetric Anal Sphincter Injuries (OASIS). It’s unclear if structural use of a mediolateral episiotomy at operative vaginal delivery will decrease this risk. The aim of our study was to evaluate the incidence of OASIS in women with operative vaginal delivery, and to assess if a mediolateral episiotomy is preventive for OASIS in these deliveries. STUDY DESIGN: A retrospective cohort study was performed using the obstetric-database of the Amphia Hospital, The Netherlands of deliveries from 2001 up to 2009. All patients with live born infants delivered by an operative vaginal delivery at a gestational age 34 weeks were included. Exclusion criteria were: multiple gestations, breech deliveries and the use of a median episiotomy. Continuous variables were compared using the Students t test or Mann-Whitney U test, the 2 test for categorical variables. Continuous variables were summarized as means with standard deviations, or medians with interquartile ranges. A logistic regression model was used for the risk assessment for OASIS when a mediolateral episiotomy was performed. Treatment effect was presented as adjusted odds ratio with 95% confidence interval. RESULTS: There were 2970 operative vaginal deliveries in the study period meeting the inclusion and exclusion criteria (Figure 1). The prevalence of OASIS was 5.7%. The absolute risk was 3.3% in the MLE group, compared to 15.5% in the MLEgroup (OR 0.19; 95% CI: 0.14-0.26). The risk estimation for OASIS remained similar (adjusted OR 0.20; 95% CI: 0.12-0.34), after logistic regression analysis with correction for gestational age, parity, birth weight, maternal age, use of epidural analgesia, indication for operative vaginal delivery, cephalic fetal position, and duration of the second stage. CONCLUSION: The use of a mediolateral episiotomy in operative vaginal delivery is associated with a fivefold decreased risk for developing OASIS. Therefore we advocate the structural use of a mediolateral episiotomy during an operative vaginal delivery in order to prevent OASIS. www.AJOG.org Intrapartum Fetal/Clinical Obstetrics Oral Concurrent Session 6

Endothelial nitric oxide synthase gene polymorphisms are not associated with recurrent aphthous stomatitis

Objective Recurrent aphthous stomatitis (RAS) is a common ulcerative disease of the oral mucosa. Recurrent oral aphthous ulceration is the most common and consistent feature of the more serious and systemic Behçet's disease (BD). Association of endothelial nitric oxide synthase (eNOS) gene polymorphisms with BD have been reported in different populations. This study aims to investigate if there is an association between these polymorphisms and RAS. Methods A case–control association study using 91 Caucasoid RAS patients and 91 ethnically matched systemically healthy controls were genotyped for the −786 and 894 eNOS single nucleotide polymorphisms (SNPs) and a variable number of tandem repeat (VNTR) polymorphism. Chi-square analysis was used to compare the allele and genotype frequencies. Results No significant difference was found in the distribution of allele and genotype frequencies of the −786 and +894 polymorphisms or the VNTR polymorphism between cases and controls. Conclusion eNOS gene polymorphisms associated with BD are not associated with RAS. This suggests that the oral ulceration common to both conditions may have a different underlying genetic aetiology. Although our data suggests that RAS does not have an association with eNOS, it is still possible that nitric oxide is involved in the disease process. This could still occur through localised and inflammation driven regulation by iNOS.

Endothelial nitric oxide synthase polymorphisms and hypertension.

The human endothelial nitric oxide synthase (eNOS) gene is highly polymorphic. Evidence for the involvement of eNOS single nucleotide polymorphisms in the development of essential hypertension is limited, though the eNOS Glu298Asp polymorphism appears to influence the blood pressure response to exercise. This variant also influences endothelial function, with its effects becoming manifest during the adaptive vascular changes of pregnancy. Carriers of eNOS Asp298 may be at risk of developing pre-eclampsia. Molecular studies have indicated that intact eNOS Asp298 has equivalent enzymatic activity to eNOS Glu298, but undergoes selective proteolysis in native cells and tissues such that the steady state level of active eNOS may be reduced in carriers of this allele. Carriers of eNOS Asp298, particularly if exposed to adverse environmental infuences on endothelial function, may be at increased risk of developing atherosclerosis and cerebrovascular disease.