Aldosterone-producing adenoma (APA) cause primary aldosteronism—the most frequent form of secondary hypertension. Somatic mutations in genes coding for ion channels and ATPases are found in APA and in aldosterone-producing cell clusters. We investigated the genetic, cellular, and molecular heterogeneity of different aldosterone-producing structures in adrenals with APA, to get insight into the mechanisms driving their development and to investigate their clinical and biochemical correlates. Genetic analysis of APA, aldosterone-producing cell clusters, and secondary nodules was performed in adrenal tissues from 49 patients by next-generation sequencing following CYP11B2 immunohistochemistry. Results were correlated with clinical and biochemical characteristics of patients, steroid profiles, and histological features of the tumor and adjacent adrenal cortex. Somatic mutations were identified in 93.75% of APAs. Adenoma carrying KCNJ5 mutations had more clear cells and cells expressing CYP11B1, and fewer cells expressing CYP11B2 or activated β-catenin, compared with other mutational groups. 18-hydroxycortisol and 18-oxocortisol were higher in patients carrying KCNJ5 mutations and correlated with histological features of adenoma; however, mutational status could not be predicted using steroid profiling. Heterogeneous CYP11B2 expression in KCNJ5 -mutated adenoma was not associated with genetic heterogeneity. Different mutations were identified in secondary nodules expressing aldosterone synthase and in independent aldosterone-producing cell clusters from adrenals with adenoma; known KCNJ5 mutations were identified in 5 aldosterone-producing cell clusters. Genetic heterogeneity in different aldosterone-producing structures in the same adrenal suggests complex mechanisms underlying APA development.

Primary aldosteronism is affecting about 10% of hypertensive patients. Primary aldosteronism should be diagnosed by screening tests based on plasma aldosterone concentration (PAC) and aldosterone-to-renin ratio (ARR), followed by confirmatory test. The cutoff values for PAC and ARR depend on PAC and plasma renin measurement methods. Liquid chromatography–tandem mass spectrometry (LC-MS/MS), the new gold standard method for aldosterone determination, is now widespread but shows lower values than immunoassays. New cutoff values have yet to be determined with LC-MS/MS PAC. In a retrospective cohort, we measured PAC by LC-MS/MS in 93 healthy volunteers, 77 patients with essential hypertension and 82 primary aldosteronism patients (42 lateralized, 24 bilateral, 16 primary aldosteronism without adrenal vein sampling) after 30 min in a seated position. PAC ranged from 42 to 309 pmol/l in healthy volunteers and from 63 to 362 pmol/l in essential hypertensive patients. A cutoff value of 360 pmol/l for basal PAC had a sensitivity of 90.5% and a specificity of 95.1% to differentiate lateralized primary aldosteronism from essential hypertensive patients. ARR ranged from 2.3 to 22.3 in healthy volunteers and from 3.2 to 55.6 pmol/mU in essential hypertensive patients. Using ROC curves, we selected an ARR of 46 pmol/mU, which provided a sensitivity of 100% and a specificity of 93.4% to distinguish between essential hypertensive and lateralized primary aldosteronism patients (sensitivity 94.4%, specificity 93.9% for the overall primary aldosteronism population). Criteria for primary aldosteronism screening need to be adapted, given the increasing use of LC-MS/MS to determine PAC. We suggest to use 360 pmol/l and 46 pmol/mU as cutoff values, respectively, for basal PAC and ARR after 30 min of seated rest.

Identifying mitotane-induced mitochondria-associated membranes dysfunctions : metabolomic and lipidomic approaches Identifying mitotane-induced mitochondria-associated membranes dysfunctions : metabolomic and lipidomic approaches

Objective: Preterm infants have relative adrenal and kidney immaturity. Recently, we linked their urine sodium loss to a hypoaldosteronism at variance with an appropriate stimulation of the renin-angiotensin system. To investigate this defective aldosterone secretion, we analyse the biosynthesis pathways of adrenal steroids in neonates according to gestational age (GA). Design: Multicentre study (Premaldo) including 152 neonates classified into three groups: group 1 (very preterm (VPT)): <33 gestational weeks (GW); group 2 (preterm (PT)): 33–36 GW and group 3 (term (T)): ≥GW. Method: Steroidomic profiles of mineralocorticoids, glucocorticoids and adrenal androgens were established from umbilical cord at birth (n=152) and peripheral blood at day 3 (n=70) using a recently developed liquid chromatography mass spectrometry method (LC-MS/MS). The enzymatic activity of each biosynthesis step was estimated by the product-to-substrate ratio. Results: At birth, VPT infants exhibit a global defect in adrenal steroid synthesis pathways leading to lower levels of aldosterone, cortisol and androstenedione than in term infants. This defect was strongly related to GA. On day 3, steroid precursors (progesterone, 11-deoxycorticosterone (DOC), 17-hydroxyprogesterone(17-OH-P) and 11-deoxycortisol (S)) were higher in VPT and negatively correlated with GA. Despite of precursors’ accumulation, aldosterone and cortisol were similar in the three groups. At birth and day 3, a low cortisol/11-deoxycortisol ratio was found in preterm infants, suggesting an 11-beta-hydroxylase activity (CYP11B1) deficiency. Conclusions: At birth, VPT infants exhibit a global deficit in mineralocorticoids, glucocorticoids and adrenal androgens that attenuates on day 3 of life. Steroid profiling using LC-MS/MS provides evidence for a partial defect in 11-hydroxylase along with prematurity.