Heterotopic Adipose Tissue in the Placental Parenchyma: Case Report

Objective: Reports of heterotopic tissue in the placenta are few and mainly include liver and adrenal cells. We report on adipose tissue found in the placenta. Case report: We present the case of a microscopic finding in a 25-year-old’s placenta who suffered from hypertensive disorder in pregnancy. During routine microscopic study, we observed a heterotopic, benign, circumscribed and intervillous nodule of adipose tissue. Conclusion: To our knowledge, there is no other reported case of adipocytes among chorionic villi. Why foreign tissues show up in the placenta remains unknown; however, several new theories offer explanations.

Autonous Aldosterone Secretion as a Subclinical Form of Primary Aldosteronism: Pathogenesis and Clinical Significance

In recent years, a substantial prevalence of primary aldosteronism (PA) has been demonstrated in both normotensive and mildly hypertensive cohorts. Consequently, a classic presentation of the syndrome, i. e. moderate-to-severe and resistant hypertension with concomitant hypokalemia, should be considered a tip-of-the-iceberg phenotype of a wide PA spectrum. Its entire range encompasses the non-classic clinical forms of mild hypertension and prehypertension but also several biochemical presentations, including patients who meet PA screening and confirmation test criteria, as well as those with either of them and those with other parameters indicating mineralocorticoid excess. In the current review, research insights on the pathogenetic background and clinical significance of autonomous aldosterone secretion (AAS) are presented, which is defined as a constellation of either: 1) normotension, normokalemia, a positive PA screening (high aldosterone-to-renin ratio) and/or confirmation test, or 2) hypertension, normokalemia and a positive PA screening but negative confirmation test. For this purpose, a literature search of the PubMed database was conducted. Advances in immunohistochemistry and genetic sequencing of isolated adrenal cells are provided as probable morphologic basis of the wide range of aldosterone secretion autonomy. Also, the role of corticotropin as an aldosterone secretagogue is discussed. To date, clinical studies depict consequences of subclinical PA phenotypes, such as increased mortality and risk of developing hypertension, impaired arterial and kidney function, association with metabolic syndrome and age, as well as osteoporosis.

Investigation of the Morphology of Adrenal Glands in Hens Kept in Two Different Housing Systems—A Pilot Study

It is difficult to objectively assess the chronic effects of housing systems on livestock and particularly on laying hens. However, this seems to be important in the context of animal welfare. Therefore, we conducted the present study in order to compare the effect of two different housing conditions, single cage (SC) and floor pen (FP), on the morphology of the adrenal gland. A higher amount of interrenal cells, which secrete stress hormones, can lead to a difference in the relation of adrenal and interrenal cells, which could be interpreted as an indication of chronic stress. For this purpose, adrenal glands were extracted, prepared, stained and examined by microscopy, and total area of the cut, total area of interrenal cells and total area of adrenal cells were measured. As a result, all laying hens had a higher percentage of interrenal cells than adrenal cells (FP: interrenal cells/adrenal cells = 78.37%/21.63%; SC: 80.00%/20.00%). The median of adrenal–interrenal ratio did not differ significantly (FP = 0.2503, SC = 0.2499), while the variation of the ratio between laying hens in FP and SC showed a slight tendency of a higher ratio in adrenal glands of FP (p < 0.0870). Body weight and adrenal–interrenal ratio were significantly negatively correlated in laying hens in FP (rS = −0.943, p < 0.0048) but not in SC (rS = −0.162, p = 0.7283). There was no significant correlation between body weight and total cell area for interrenal cells or adrenal cells. Body weight was significantly lower for laying hens kept in SC than for laying hens kept in FP (p < 0.0001). Due to the present results, it can be concluded that keeping laying hens in single cages can have a negative effect on body weight.

The Herbicide Atrazine Potentiates Angiotensin II-Induced Aldosterone Synthesis and Release From Adrenal Cells

Atrazine is one of the most commonly used pre-emergence and early post-emergence herbicides in the world. We have shown previously that atrazine does not directly stimulate the pituitary or adrenal to trigger hormone release but acts centrally to activate a stress-like activation of the hypothalamic-pituitary-adrenal axis. In doing so, atrazine treatment has been shown to cause adrenal morphology changes characteristic of repeated stress. In this study, adrenals from atrazine treated and stressed animals were directly compared after 4 days of atrazine treatment or restraint stress. Both atrazine and stressed animals displayed reduced adrenocortical zona glomerulosa thickness and aldosterone synthase (CYP11B2) expression, indicative of repeated adrenal stimulation by adrenocorticotropic hormone. To determine if reduced CYP11B2 expression resulted in attenuated aldosterone synthesis, stressed and atrazine treated animals were challenged with angiotensin II (Ang II). As predicted, stressed animals produced less aldosterone compared to control animals when stimulated. However, atrazine treated animals had higher circulating aldosterone concentrations compared to both stressed and control groups. Ang II-induced aldosterone release was also potentiated in atrazine pretreated human adrenocortical carcinoma cells (H295R). Atrazine pretreated did not alter the expression of the rate limiting steroidogenic StAR protein or angiotensin II receptor 1. Atrazine treated animals also presented with higher basal blood pressure than vehicle treated control animals suggesting sustained elevations in circulating aldosterone levels. Our results demonstrate that treatment with the widely used herbicide, atrazine, directly increases stimulated production of aldosterone in adrenocortical cells independent of expression changes to rate limiting steroidogenic enzymes.

CTNNB1-Mutant Aldosterone-Producing Adenomas With Somatic Mutations of GNA11/GNAQ Have Distinct Phenotype and Genotype

Background: We report (this meeting) somatic mutation of GNA11/Q in CTNNB1-mutant APAs. The recurrent co-driver mutation causes reversible hypertension in puberty, pregnancy, or menopause. We have investigated the molecular mechanism of this association. Methods: Gene expression profiles in 3 double mutant APAs were studied by unsupervised hierarchical clustering analysis and enrichment analysis of 362 differentially expressed genes and validated by qPCR, IFC and IHC in 10 double mutant APAs or transfected primary adrenal cells. Multiple region biopsies were performed in 7 adrenals adjacent to double-mutant APAs and 4 APAs with KCNJ5 or CACNA1D mutations. The findings of APA mutations in adjacent adrenals were replicated in each case by ddPCR ± NGS. Results: Unsupervised hierarchical clustering analysis showed clustering of the double-mutant APAs, and a high proportion of genes were many-fold upregulated compared to other APAs. LHCGR, TMEM132E, DKK1, C9orf84, FAP, GNRHR and MPP3 are among the genes with high expression. A small number of genes are down-regulated in the double-mutant APAs, including CYP11B1. qPCR confirmed an average of ~10 to 1000-fold higher expression of the hallmark genes in double-mutants. Enrichment analysis showed significant enrichment of features or terms concerned with cell junction and cell adhesion (P&lt;10–8). IFC confirmed LHCGR intensity was 31–144 fold higher in primary adrenal cells with GNA11-p.Gln209Pro transfection and high CTNNB1 intensity. LHCGR intensity was qualitatively and quantitatively associated with immunofluorescence for CTNNB1. IHC of double-mutant APAs showed absent CYP11B1 but strong staining of CYP11B2. qPCR confirmed a lower CYP11B1/CYP11B2 ratio and a higher LHCGR expression (P&lt;10–3, both). IHC confirmed LHCGR positivity in double-mutant APAs but distribution varied both within and between cells. Adjacent ZG was hyperplastic, with absence of both CYP11B1 and CYP11B2 staining, but weak/moderate staining for LHCGR. The same GNA11 ± CTNNB1 somatic mutations were detected in multiple regions of the adjacent adrenals to 3 double mutant APAs. qPCR of hallmark APA genes differed from the APAs. High concordance between ddPCR, NGS and Sanger sequencing of the findings of APA mutations in adjacent adrenals when analysed in the same sample. No mutations were found in 4 adrenals adjacent to APAs with KCNJ5 or CACNA1D mutations, nor in other 4 adrenals adjacent to double-mutant APAs. Conclusions: Patients harboring APAs with somatic mutations in both GNA11/GNAQ Q209 and CTNNB1 have distinct phenotype in both the APAs and their adjacent adrenals. Same GNA11 ± CTNNB1 somatic mutations were found in the adjacent adrenals to double mutant APAs. We infer that a double-hit within related pathways is more likely than a single-hit to cause large increases in expression of LHCGR, and of other genes which may influence clinical presentation.

Gender difference in adrenal developmental toxicity induced by dexamethasone and its intrauterine programming mechanism

Background and Purpose: Dexamethasone is widely used in preterm labor and related diseases. However, prenatal dexamethasone exposure (PDE) can cause multi-organ developmental toxicities in offspring. Our previous study found the occurrence of fetal-originated diseases were associated with adrenal developmental programming alteration in offspring. Here, we investigated the effects of PDE on the adrenal function in offspring and its intrauterine programming mechanism. Experimental Approach: A rat model of PDE was established to observe the alteration of adrenal steroidogenesis in offspring. Further, we confirmed the gender difference of adrenal steroidogenesis and its molecular mechanism combined with in vivo and in vitro experiment. Key Results: PDE caused a decrease in adrenal steroidogenic function in fetal rats, but decreased in males and increased in females after birth. Meanwhile, the adrenal H3K14ac level and expression of 11β-hydroxysteroid dehydrogenase 2 (11β-HSD2) in PDE offspring were decreased in males and increased in females, suggesting 11β-HSD2 might mediate gender difference of adrenal function. We further confirmed dexamethasone inhibited the H3K14ac level and expression of 11β-HSD2 through GR/SP1/p300 pathway. After bilateral testectomy or ovariectomy in adult PDE offspring rats, adrenal 11β-HSD2 expression and steroidogenic function were both reduced. Using rat primary fetal adrenal cells, the differential expression in AR and ERβ were proved to involve in regulating the gender difference of 11β-HSD2 expression. Conclusion and Implications: This study demonstrated the gender difference in adrenal steroidogenic function of PDE offspring after birth, and elucidates a sex hormone receptor-dependent epigenetically regulating mechanism for adrenal 11β-HSD2 programming alteration.

BEX1 Is Differentially Expressed in Aldosterone-Producing Adenomas and Protects Human Adrenocortical Cells From Ferroptosis

Aldosterone-producing adenomas (APAs) are a major cause of primary aldosteronism. Somatic mutations in ion channels and transporters drive the aldosterone overproduction in the majority of APAs with mutations in the KCNJ5 G protein-coupled potassium channel predominating in most reported populations. Our objective was to gain insight into biological mechanisms of APA tumorigenesis by comparing transcriptomes of APAs of distinct sizes by mRNA sequencing analysis (9 APAs with adenoma diameter ≥30 mm versus 12 APAs ≤10 mm). Genes with significantly altered expression levels between these 2 groups were identified in APAs with no mutation detected (348 genes) and with a KCNJ5 mutation (155 genes). We validated the differential expression of 10 genes with a known function related to cell death and proliferation in an expanded sample set of 71 APAs by real-time quantitative polymerase chain reaction (58 macro-APAs, diameter ≥10 mm; 13 micro-APAs, diameter <10 mm). We focused on BEX1 that was upregulated in micro-APAs relative to macro-APAs (2.76-fold, P <0.001) and compared with paired adrenal cortex (3.85-fold, P <0.05), and showed a linear negative correlation with APA diameter in the no mutation detected group (r=−0.501, P =0.007). Compared with control cells, stable expression of BEX1 in human adrenocortical cells did not alter cell cycle progression or sensitivity to apoptosis but conferred protection from ferroptosis ( P <0.01), a form of regulated cell death, measured by flow cytometry. Taken together, these findings demonstrate that BEX1 promotes cell survival in adrenal cells by mediating the inhibition of ferroptosis and suggest a function for BEX1 in the pathogenesis of APAs.