Primary aldosteronism (PA) is characterized by the onset of both cardiac and gluco-metabolic alterations. The aim of this study was to evaluate the impact of aldosterone excess on the development of such complications, and the effects of surgical and pharmacological treatment on their long-term outcome. We prospectively re-examined 61 patients: 25 with aldosterone-producing adenoma (APA), after surgery, and 36 patients with idiopathic hyperaldosteronism (IHA) on pharmacological treatment. The lipid, fasting and dynamic glucose profiles and the echocardiographic parameters were evaluated at diagnosis and at follow-up. After adrenalectomy all patients had normalization of aldosterone levels and were cured of hypokalaemia, and a resolution of hypertension was achieved in 12 of 25 patients. APA patients showed a significant reduction of both plasma glucose (P = 0.017) and insulin levels (P = 0.001) after 75 g oral glucose tolerance test. Stabilization of glucose metabolism complications was observed in IHA patients. Multiple regression analysis at diagnosis showed a positive correlation between homeostasis model assessment (HOMA) insulin resistance index and HOMA β cell and serum aldosterone levels in both APA and IHA. Echocardiographic parameters were improved in both APA and IHA at follow-up and the difference was statistically significant for left ventricular mass index (P = 0.017) and interventricular septum thickness (P = 0.007) in APA patients. The removal of aldosterone excess in APA patients induces the regression of both cardiac and gluco-metabolic complications, indicating aldosterone as a main determinant of such alterations. In IHA patients the medical treatment seems to avoid the possible progression of the these alterations that appear to be stable.

Aldosterone-producing adenomas (APAs) cause a sporadic form of primary aldosteronism and somatic mutations in the KCNJ5 gene, which encodes the G-protein–activated inward rectifier K ⁺ channel 4, GIRK4, account for ≈40% of APAs. Additional somatic APA mutations were identified recently in 2 other genes, ATP1A1 and ATP2B3 , encoding Na ⁺ /K ⁺ -ATPase 1 and Ca ²⁺ -ATPase 3, respectively, at a combined prevalence of 6.8%. We have screened 112 APAs for mutations in known hotspots for genetic alterations associated with primary aldosteronism. Somatic mutations in ATP1A1 , ATP2B3 , and KCNJ5 were present in 6.3%, 0.9%, and 39.3% of APAs, respectively, and included 2 novel mutations (Na ⁺ /K ⁺ -ATPase p.Gly99Arg and GIRK4 p.Trp126Arg). CYP11B2 gene expression was higher in APAs harboring ATP1A1 and ATP2B3 mutations compared with those without these or KCNJ5 mutations. Overexpression of Na ⁺ /K ⁺ -ATPase p.Gly99Arg and GIRK4 p.Trp126Arg in HAC15 adrenal cells resulted in upregulation of CYP11B2 gene expression and its transcriptional regulator NR4A2. Structural modeling of the Na ⁺ /K ⁺ -ATPase showed that the Gly99Arg substitution most likely interferes with the gateway to the ion binding pocket. In vitro functional assays demonstrated that Gly99Arg displays severely impaired ATPase activity, a reduced apparent affinity for Na ⁺ activation of phosphorylation and K ⁺ inhibition of phosphorylation that indicate decreased Na ⁺ and K ⁺ binding, respectively. Moreover, whole cell patch-clamp studies established that overexpression of Na ⁺ /K ⁺ -ATPase Gly99Arg causes membrane voltage depolarization. In conclusion, somatic mutations are common in APAs that result in an increase in CYP11B2 gene expression and may account for the dysregulated aldosterone production in a subset of patients with sporadic primary aldosteronism.

The upright serum aldosterone/upright plasma renin activity ratio (ARR) has been recommended as a screening tool for the diagnosis of primary aldosteronism. We reviewed the data collected from hypertensive patients in order to define retrospectively the cut-off values and evaluate the reliability of the ARR and of the saline infusion test in the diagnosis of primary aldosteronism. In 157 patients referred to our unit with a suspicion of primary aldosteronism, 61 of whom had confirmed primary aldosteronism [26 aldosterone-producing adenoma (APA); 35 idiopathic hyperaldosteronism], the supine and upright ARR, and the ARR after the administration of captopril and losartan were calculated, and the results of the saline infusion test were analysed. Choosing 40 as the cut-off value, the upright ARR had 100% sensitivity and 84.4% specificity. The post-captopril and post-losartan ARR were slightly more specific, but at the cost of a lower sensitivity. A cut-off value of 7 ng/dl for serum aldosterone at the end of the saline infusion in patients with an upright ARR of 40, gave 100% specificity and a positive predictive value. Furthermore, APA patients showed increased mean levels of aldosterone/cortisol ratio after the saline infusion test. Our data reinforce the superiority of a standardized upright ARR as a screening test in the diagnosis of primary aldosteronism, identifying 40 as an ideal cut-off value. Saline infusion represents a useful test to confirm such a diagnosis, with a serum aldosterone level of 7 ng/dl as a satisfactory cut-off value. Some more information is obtained when the aldosterone/cortisol ratio is considered.