Haplotypes of angiotensinogen in essential hypertension.

The M235T polymorphism of the angiotensinogen gene (AGT) has been associated with essential and pregnancy-induced hypertension. Generation of haplotypes can help to resolve whether the T235 allele itself predisposes to the development of hypertension or acts as a marker of an unknown causal molecular variant. We identified 10 diallelic polymorphisms at the AGT locus and genotyped both a series of 477 probands of hypertensive families and 364 controls, all French Caucasians, as well as a series of 92 hypertensives and 122 controls from Japan. Despite a large ethnic difference in gene frequency, a significant association of T235 with hypertension was observed both in Cancasians (.46 vs. .38, P = .004) and in Japanese (.91 vs. .76, P = .002). In both groups, the G-->A substitution located at position -6 upstream of the initial transcription site occurred at the same frequency and in complete linkage disequilibrium with the T235 allele. No other polymorphism was found to be consistently associated with hypertension. Five informative haplotypes subdividing the T235 allele were generated. Whereas two of them were associated with hypertension in Caucasians, none of these two haplotypes (H3 and H4) reached statistical significance in Japanese. The analysis of the AGT-GT repeat revealed marked linkage disequilibriums between each of the diallelic polymorphisms and some (GT)n alleles, with similar patterns in the two populations. The strong disequilibrium between M235 and (GT)16 explained the increased frequency of that particular allele in French controls compared with hypertensives (.42 vs. .36, P < .01). The haplotype combining the M235T and G-6A polymorphisms appears as the ancestral allele of the human AGT gene and as the one associated with hypertension.

[Hyperaldosteronism sensitive to dexamethasone with adrenal adenoma. Clinical, biological and genetic study]. / Hyperaldostéronisme sensible à la dexaméthasone avec adénome surrénalien. Etude clinique, biologique et génétique.

OBJECTIVES: Dexamethasone-sensitive hyperaldosteronism is associated with early onset hypertension and primary hyperaldosteronism. Diagnosis is difficult but can be improved by genetic testing for the mutant gene. METHODS: We collected the clinical, biological and genetic elements observed in a family with dexamethasone-sensible hyperaldosteronism. Complete data were obtained in 5 adult subjects with the disease. Degree of hypertension varied, more so in the second generations as did hypokaliaemia and hyperaldosteronism. In affected patients, there was a 10 to 50 fold increase in urinary 18-OH components and 18 oxocortisol. RESULTS: Single dose (1.5 mg) dexamethasone led to a greater than 80% drop in aldosterone levels in the blood and urine, confirming the abnormal effect of ACTH on mineralocorticoid secretion. At the dose of 1 mg/d for 10 weeks, dexamethasone lowered mean 24-H ambulatory arterial pressure (11.8/9.6 mmHg) and corrected for the hypokaliaemia (+0.54 mmol/l) and the hyperaldosteronism (mean decrease -36% and -75% in blood and urine respectively). An adrenal tumour was identified in hyperplasic glands in two subjects and a micronodular formation was identified in two others. The specific molecular diagnosis of the disease was done with Southern blotting. Among the 18 families in 3 generations, 8 carried a 11 beta OHase-aldosterone synthetase chimeric gene. This mutation cosegregates with hormonal abnormalities and confirms the autosomal dominant inheritance of the disease. CONCLUSION: The simplicity and rapidity of genetic testing allows early diagnosis of this disease among families with early onset hypertension and associated hyperaldosteronism with or without adrenal hyperplasia and/or a tumoral formation.

Lack of evidence for linkage of the endothelial cell nitric oxide synthase gene to essential hypertension.

BACKGROUND: The basal release of nitric oxide by the endothelium plays an important role in regulating blood flow and pressure and mediates most of the endothelium-dependent vasodilation. Impairment of nitric oxide production by specific inhibitors increases blood pressure in humans, and several reports suggest that hypertensive subjects have a blunted endothelium-dependent vasodilatation that might be secondary to decreased nitric oxide production from the vessel wall. METHODS AND RESULTS: To determine whether the endothelial nitric oxide synthase gene is involved in human essential hypertension, we identified informative biallelic and multiallelic markers of this locus and performed case-control and linkage studies in hypertensive subjects and normotensive control subjects. We used the affected sib pair method to test for potential linkage in 145 hypertensive pedigrees (269 sib pairs, 346 subjects) with a highly polymorphic marker of the nitric oxide synthase gene (polymorphism information content of 92%). There was no evidence for linkage among affected siblings. The 95% upper confidence limit of this value suggests that at most 1% of alleles in excess of expected are shared. We also identified two informative biallelic markers of this gene to perform a case-control study on white hypertensive and normotensive subjects. Similar genotype distributions between the two groups were noted for both markers. Estimated haplotype frequencies by maximum likelihood methods combining the two biallelic markers were also similar in both groups. CONCLUSIONS: These findings do not suggest that common molecular variants of the endothelial nitric oxide synthase gene are involved in essential hypertension.

Short report: HYPERGENE: a clinical and genetic database for genetic analysis of human hypertension.

OBJECTIVE: Genetic studies of essential human hypertension require the recording and management of numerous data concerning multiple hypertensive families. The present report describes a new family database, HYPERGENE, and demonstrates its potential usefulness in such a complex disease. METHODS: The database was implemented on an Apple Macintosh computer using the 4TH DIMENSION software program. Through a user-friendly interface, it offers a high-quality data record, easy data retrieval and compatibility with other software. PATIENTS: HYPERGENE contains a prospective collection of 187 families with at least two hypertensive sibs (826 subjects). Each subject was analysed according to the same protocol. To allow definition of clinical and biological phenotypes and genetic analysis, clinical and biological data were recorded and, at the same time, plasma, urine and DNA libraries were stored. RESULTS: Probands were 50.6 years old with an early onset (39.1 years of age) of hypertension (157.7/97.8 mmHg); 125 had moderate-to-severe hypertension. According to our selection criteria, only one out of 187 sibships had familial dislipidaemic hypertension. Of the living fathers, 45% were examined, and 54% of the living mothers: 48.6% had an onset of hypertension before age 50 years. Children (mean age 28.7 years) of hypertensive sibs presented a high percentage of hypertension (8.3%). CONCLUSION: The HYPERGENE database facilitates data storage and analysis on familial hypertension, and should prove a useful tool for assessing molecular biology results in the field of hypertension and for allowing collaborative research.

Angiotensin II type 1 receptor gene polymorphisms in human essential hypertension.

We conducted the present study to determine whether the angiotensin II type I receptor (AT1) gene might be implicated in human essential hypertension by using case-control and linkage studies. The entire coding and 3' untranslated regions of the AT1 receptor gene (2.2 kb) were amplified by polymerase chain reaction and submitted to single-strand conformation polymorphism in 60 hypertensive subjects with a familial susceptibility. We identified five polymorphisms (T573-->C, A1062-->G, A1166-->C, G1517-->T, and A1878-->G). However, no mutations that alter the encoded amino acid sequence were detected. A case-control study performed on white hypertensive (n = 206; blood pressure, 168 +/- 16/103 +/- 9 mm Hg) and normotensive (n = 298; blood pressure, 122 +/- 10/75 +/- 9 mm Hg) subjects using three of five polymorphisms showed a significant increase in allelic frequency of C1166 in hypertensive subjects (0.36 versus 0.28 for normotensive subjects, chi 2 = 6.8, P < .01). Frequencies for the alleles of the other two polymorphisms (T573-->C, A1878-->G) were similar in both groups. We performed a linkage study using the affected sib pair method and a highly polymorphic marker of the AT1 receptor gene. There was no evidence for linkage in 267 sib pairs analyzed from 138 pedigrees. These findings would be compatible with a common variant of the AT1 receptor imparting a small effect on blood pressure; further studies will be needed to address this possibility.

HYPERGENE: a clinical and genetic database for genetic analysis of human hypertension.

OBJECTIVE: Genetic studies of essential human hypertension require the recording and management of numerous data concerning multiple hypertensive families. The present paper describes a new family database, HYPERGENE, and demonstrates its potential usefulness in such a complex disease. METHODS: The database was implemented on an Apple Macintosh computer using the 4TH DIMENSION software program. Through a user-friendly interface, it offers a high-quality data record, easy data retrieval and compatibility with other software. PATIENTS: HYPERGENE contains a prospective collection of 187 families with at least two hypertensive sibs (826 subjects). Each subject was analysed according to the same protocol. To allow definition of clinical and biological phenotypes and genetic analysis, clinical and biological data were recorded and, at the same time, plasma, urine and DNA libraries were stored. RESULTS: Probands were 50.6 years old with an early onset (39.1 years of age) of hypertension (157.7/97.8 mmHg); 125 had moderate-to-severe hypertension. According to our selection criteria, only one out of 187 sibships had familial dislipidaemic hypertension. Of the living fathers, 45% were examined, and 54% of the living mothers: 48.6% had an onset of hypertension before age 50 years. Children (mean age 28.7 years) of hypertensive sibs presented a high percentage of hypertension (8.3%). CONCLUSION: The HYPERGENE database facilitates data storage and analysis on familial hypertension, and should prove a useful tool for assessing molecular biology results in the field of hypertension and for allowing collaborative research.

Can the genetic factors influence the treatment of systemic hypertension? The case of the renin-angiotensin-aldosterone system.

The hereditary nature of familial hypertension has been clearly established by a number of clinical studies. About 30% of the blood pressure variance can be attributed to genetic factors. As a consequence, the relative risk for developing coronary artery disease or cardiovascular death is increased in patients with a family history of hypertension and cardiovascular disease. Patients with such familial history should be considered at the same risk as those who have independent epidemiologic risk factors. The development of molecular genetics allows establishment of a link between high blood pressure, intermediate phenotypes, and the genes involved in blood pressure regulation. Gene markers should be available in the near future that will help to identify patients predisposed to hypertension. The genes of the renin-angiotensin-aldosterone system are good examples of candidate genes whose products are known to participate in blood pressure regulation. The possible involvement of these genes in essential hypertension is critically analyzed.

Molecular basis of human hypertension: role of angiotensinogen.

Essential hypertension is a common human disease believed to result from the interplay of multiple genetic and environmental determinants. In genetic studies of two large panels of hypertensive sibships from widely separated geographical areas, we obtained evidence of genetic linkage between the angiotensinogen gene (AGT) and hypertension, demonstrated association of AGT molecular variants with the disease, and found significant differences in plasma concentrations of angiotensinogen among hypertensive subjects with different AGT genotypes. The corroboration and replication afforded by these results support the interpretation that molecular variants of AGT constitute inherited predispositions to essential hypertension in humans.

Sib pair linkage analysis of renin gene haplotypes in human essential hypertension.

Although essential arterial hypertension is believed to have a strong genetic predisposition, the gene(s) responsible are unknown. The mechanisms underlying the regulation of blood pressure and experimental studies place the renin gene among the main candidate genes that need to be tested in humans. We tested the hypothesis of a linkage between the renin gene and essential hypertension using the affected sib pair method. Siblings (133 subjects, 52.1 +/- 10.9 years) from 57 families were selected for sustained hypertension (160.7 +/- 22.9/99.5 +/- 12.8 mmHg with 80% of patients under antihypertensive treatment), of early onset (40.7 +/- 12.0 years), in the absence of obesity, diabetes mellitus, and secondary hypertension. Eight renin haplotypes were generated from three diallelic renin restriction fragment length polymorphisms (RFLPs) (TaqI, HinfI, HindIII) located throughout the renin gene. The allelic concordance between the sib pairs was analyzed by identity by state relationships for 98 sib pairs (41 for 41 couples, 39 for 13 trios, 18 for 3 quartets). Allelic frequencies in the 57 hypertensive probands were similar to those observed among 102 hypertensive subjects studied previously. Six of eight possible haplotypes were observed, the informativity of the marker corresponded to 70% of heterozygosity. Allelic concordance for all sib pairs according to sibship size was not significantly different from that expected under the hypothesis of no linkage (t = 0.52, P = 0.15) reflecting only a small excess of renin alleles shared by the hypertensive sibs (1.44 +/- 0.6 vs 1.36 +/- 0.6). Likewise the linkage hypothesis was unsupported by weighted estimates to correct for possible bias due to large sibship size. Thus, the sib pair analysis suggests that the renin gene does not have a frequent role in the pathogenesis of essential hypertension; further more powerful linkage studies or other approaches will be needed to detect contributions at the renin locus to the heritability of essential hypertension.

Long-term metabolic effects of spironolactone and thiazides combined with potassium-sparing agents for treatment of essential hypertension.

By using information prospectively collected in the computerized ARTEMIS databank, the long-term metabolic consequences of spironolactone, hydrochlorothiazide-amiloride combination and cyclothiazide-triamterene combination were evaluated in 100 patients for each group matched according to sex, age and blood pressure (BP). Spironolactone was prescribed at a mean dose of 98 mg, hydrochlorothiazide at 36 mg and cyclothiazide at 2 mg, during a mean follow-up of 20 months. Compared with the pretherapeutic values, BP decreased equally in both treatment groups (18/9 mm Hg on average). Creatinine increased significantly in the 3 groups (9, 8, 14 mumol/liter, p less than 0.001) as did uric acid (18, 31, 42 mumol, p less than 0.001). Plasma potassium increased with spironolactone (0.7 mmol/liter, p less than 0.001) and remained stable with the combinations of the thiazide-potassium-sparing agents. For the 3 groups, the slight and nonsignificant variations of fasting blood glucose and cholesterol were mainly the results of a phenomenon of regression to the mean. However, when both groups of thiazide-treated patients were considered, the reduction of plasma potassium was accompanied by a slight increase in glucose (0.1 mmol/liter) and cholesterol levels (0.2 mmol/liter) compared with when kalemia decreased (-0.1 and -0.1 mmol/liter, respectively, p less than 0.05 and p less than 0.05). It is concluded that in a clinical daily practice of a hypertension clinic low doses of spironolactone or of thiazides combined with potassium-sparing agents reduced BP without alteration in lipid or carbohydrate metabolism on long-term follow-up.

Efficacy and tolerance of spironolactone in essential hypertension.

The long-term efficacy and tolerance of spironolactone in essential hypertension was evaluated among 20,812 patients referred to the Broussais and St. Joseph systemic hypertension clinics between 1976 and 1985 by using information prospectively collected in the computerized ARTEMIS data bank. In 182 patients (51 men, 131 women) treated with spironolactone alone during a mean follow-up period of 23 months, a mean dose of 96.5 mg decreased systolic and diastolic blood pressure (BP) by 18 and 10 mm Hg, respectively, below pretherapeutic levels. The BP decrease was greater with doses of 75 to 100 mg (12.4% and 12.2%) than with doses of 25 to 50 mg (5.3 and 6.5%, p less than 0.001), but no additional decrease was found with doses above 150 mg. Plasma creatinine level increased modestly (8.3 mumol/liters), as did plasma potassium level (0.6 mmol/liters) (both p less than 0.001); uric acid level increased, but not significantly (10.5 mumol/liter). Fasting blood glucose and total cholesterol levels did not change, triglyceride levels increased slightly (0.1 mmol/liter, p less than 0.05). These changes were similar in both sexes and were not influenced by length of follow-up. Among the 699 men prescribed spironolactone alone or in association with another antihypertensive treatment, 91 cases of gynecomastia developed (13%). Gynecomastia was reversible and dose-related; at doses of 50 mg or less the incidence was 6.9%, but 52.2% for doses of 150 mg or higher. Despite limitations inherent in the interpretation of data banks, it is concluded that spironolactone administered in daily practice reduced BP without inducing adverse metabolic adverse effects and that in patients with essential hypertension, doses should be kept below 100 mg.