Colocalization of Wnt/β-catenin and ACTH signaling pathways and paracrine regulation in aldosterone producing adenoma

2021 ◽  
Author(s):  
Kelly De Sousa ◽  
Alaa B Abdellatif ◽  
Isabelle Giscos-Douriez ◽  
Tchao Meatchi ◽  
Laurence Amar ◽  
...  
Keyword(s):  
Mast Cells ◽  
Zona Glomerulosa ◽  
Nerve Fibers ◽  
Molecular Characteristics ◽  
Paracrine Regulation ◽  
Sequence Of Events ◽  
Cellular Environment ◽  
Mutational Status ◽  
Aldosterone Producing Adenoma ◽  
And Function

Abstract Context Aldosterone-producing adenomas (APA) are a common cause of primary aldosteronism. Despite the discovery of somatic mutations in APA and characterization of multiple factors regulating adrenal differentiation and function, the sequence of events leading to APA formation remains to be determined. Objective We investigated the role of Wnt/β-catenin and ACTH signaling, as well as elements of paracrine regulation of aldosterone biosynthesis in adrenals with APA and their relationship to intratumoral heterogeneity and mutational status. Design We analyzed expression of CYP11B2, CYP17A1, β-catenin, MC2R, pCREB, Tryptase, S100, CD34 by multiplex immunofluorescence and IHC guided RT-qPCR. Setting 11 adrenals with APA and one with micronodular hyperplasia from patients with PA were analysed. Main Outcome Measure(s) Localization of CYP11B2, CYP17A1, β-catenin, MC2R, pCREB, Tryptase, S100, CD34 in APA and aldosterone producing cell clusters (APCC). Results Immunofluorescence revealed abundant mast cells and a dense vascular network in APA, independent of mutational status. Within APA, mast cells were localized in areas expressing CYP11B2 and were rarely co-localized with nerve fibers, suggesting that their degranulation is not controlled by innervation. In these same areas, ß-catenin was activated, suggesting a zona glomerulosa cell identity. In heterogeneous APA with KCNJ5 mutations, MC2R and VEGFA expression was higher in areas expressing CYP11B2. A similar pattern was observed in APCC, with high expression of CYP11B2, activated β-catenin, and numerous mast cells. Conclusions Our results suggest that aldosterone producing structures in adrenals with APA share common molecular characteristics and cellular environment, despite different mutation status, suggesting common developmental mechanisms.

scholarly journals Interaction Between Wnt/β-catenin and ACTH Signaling Pathways and Paracrine Regulation in Aldosterone Producing Adenoma

2021 ◽  
Vol 5 (Supplement_1) ◽  
pp. A290-A291
Author(s):  
Alaa B Abdellatif ◽  
Kelly De Sousa ◽  
Isabelle Giscos-Douriez ◽  
Tchao Meatchi ◽  
Laurence Amar ◽  
...  
Keyword(s):  
Mast Cells ◽  
Signaling Pathways ◽  
Somatic Mutations ◽  
Nerve Fibers ◽  
Molecular Heterogeneity ◽  
Mutation Status ◽  
Aldosterone Synthase ◽  
Paracrine Regulation ◽  
Mutational Status ◽  
Aldosterone Producing Adenoma

Abstract Primary aldosteronism (PA) is the most frequent form of secondary arterial hypertension and is caused in the majority of cases by an aldosterone producing adenoma (APA) or bilateral adrenal hyperplasia. Different somatic mutations have been identified in APA and in other aldosterone producing structures, which can be distinct within the same adrenal, suggesting multiple mechanisms underlying APA development. Also, APA show important cellular and molecular heterogeneity which may be due to interaction of different signaling pathways involved in adrenal cortex cell differentiation and function. The aim of this study was to investigate the role of Wnt/β-catenin and ACTH signaling as well as elements of paracrine regulation of aldosterone biosynthesis and vascularization in the development of APA and aldosterone producing cell clusters (APCC) and their relationship with intratumoral heterogeneity and mutational status. We performed immunohistochemistry and multiplex immunofluorescence (CYP11B2, CYP17A1, β-catenin, MC2R, pCREB, Tryptase, S100, CD34) multispectral image analysis on 11 adrenals with APA and one with micronodular hyperplasia from patients with PA. CYP11B2 (aldosterone synthase) IHC guided RT-qPCR was performed on RNA extracted from formalin-fixed paraffin-embedded tissues in 7 adrenals. Multiplex immunofluorescence revealed high abundance of tryptase positive mast cells and a dense vascular component in APA, which were independent of the mutational status. Within APA, mast cells were mainly localized in zones expressing CYP11B2, but not in areas expressing CYP17A1, and were rarely colocalized with nerve fibers, suggesting that their activity is not controlled by innervation. In cells expressing aldosterone synthase, β-catenin was activated, i.e. shows nuclear and/or cytoplasmic staining, features suggestive of a zona glomerulosa cell identity; MC2R was found at the cell membrane. Expression of MC2R mRNA was observed at different levels in APA, similar to expression of MRAP and VEGFA; MRAP2 was not detected. Within heterogeneous APA carrying KCNJ5 mutations, both MC2R and VEGFA expression was higher in areas expressing CYP11B2. Remarkably, this pattern was maintained in APCC, where cells show high CYP11B2 expression, together with activated β-catenin, independently of the mutation status. In addition, a high number of mast cells was detected around APCC, with a reorganization of the capillaries around the CYP11B2 positive cells. Our results suggest that aldosterone producing structures in adrenals with APA share common molecular characteristics and cellular environment, despite different mutation status. Mast cells appear to be closely associated with cells expressing aldosterone synthase, both in APA and APCC, and their role in regulating aldosterone biosynthesis in the context of somatic mutations in PA remains to be established.

scholarly journals Molecular characteristics of the KCNJ5 mutated aldosterone-producing adenomas

2017 ◽  
Vol 24 (10) ◽  
pp. 531-541 ◽  
Author(s):  
Masanori Murakami ◽  
Takanobu Yoshimoto ◽  
Kazuhiko Nakabayashi ◽  
Yujiro Nakano ◽  
Takahiro Fukaishi ◽  
...  
Keyword(s):  
Gene Expression ◽  
Gene Expression Omnibus ◽  
Zona Glomerulosa ◽  
Adrenal Tumors ◽  
Molecular Characteristics ◽  
Molecular Heterogeneity ◽  
Dna Hypomethylation ◽  
Mutational Status ◽  
Genome Wide ◽  
Genome Wide Gene Expression

The pathophysiology of aldosterone-producing adenomas (APAs) has been investigated via genetic approaches and the pathogenic significance of a series of somatic mutations, including KCNJ5, has been uncovered. However, how the mutational status of an APA is associated with its molecular characteristics, including its transcriptome and methylome, has not been fully understood. This study was undertaken to explore the molecular characteristics of APAs, specifically focusing on APAs with KCNJ5 mutations as opposed to those without KCNJ5 mutations, by comparing their transcriptome and methylome status. Cortisol-producing adenomas (CPAs) were used as reference. We conducted transcriptome and methylome analyses of 29 APAs with KCNJ5 mutations, 8 APAs without KCNJ5 mutations and 5 CPAs. Genome-wide gene expression and CpG methylation profiles were obtained from RNA and DNA samples extracted from these 42 adrenal tumors. Cluster analysis of the transcriptome and methylome revealed molecular heterogeneity in APAs depending on their mutational status. DNA hypomethylation and gene expression changes in Wnt signaling and inflammatory response pathways were characteristic of APAs with KCNJ5 mutations. Comparisons between transcriptome data from our APAs and that from normal adrenal cortex obtained from the Gene Expression Omnibus suggested similarities between APAs with KCNJ5 mutations and zona glomerulosa. The present study, which is based on transcriptome and methylome analyses, indicates the molecular heterogeneity of APAs depends on their mutational status. Here, we report the unique characteristics of APAs with KCNJ5 mutations.

scholarly journals Pathophysiological Roles of Neuro-Immune Interactions between Enteric Neurons and Mucosal Mast Cells in the Gut of Food Allergy Mice

Cells ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1586
Author(s):  
Tomoe Yashiro ◽  
Hanako Ogata ◽  
Syed Faisal Zaidi ◽  
Jaemin Lee ◽  
Shusaku Hayashi ◽  
...  
Keyword(s):  
Food Allergy ◽  
Mast Cells ◽  
Imaging System ◽  
Receptor Gene ◽  
Nerve Fibers ◽  
Enteric Neurons ◽  
Food Allergies ◽  
Adenosine A3 Receptor ◽  
Myenteric Neurons ◽  
Mucosal Mast Cells

Recently, the involvement of the nervous system in the pathology of allergic diseases has attracted increasing interest. However, the precise pathophysiological role of enteric neurons in food allergies has not been elucidated. We report the presence of functional high-affinity IgE receptors (FcεRIs) in enteric neurons. FcεRI immunoreactivities were observed in approximately 70% of cholinergic myenteric neurons from choline acetyltransferase-eGFP mice. Furthermore, stimulation by IgE-antigen elevated intracellular Ca2+ concentration in isolated myenteric neurons from normal mice, suggesting that FcεRIs are capable of activating myenteric neurons. Additionally, the morphological investigation revealed that the majority of mucosal mast cells were in close proximity to enteric nerve fibers in the colonic mucosa of food allergy mice. Next, using a newly developed coculture system of isolated myenteric neurons and mucosal-type bone-marrow-derived mast cells (mBMMCs) with a calcium imaging system, we demonstrated that the stimulation of isolated myenteric neurons by veratridine caused the activation of mBMMCs, which was suppressed by the adenosine A3 receptor antagonist MRE 3008F20. Moreover, the expression of the adenosine A3 receptor gene was detected in mBMMCs. Therefore, in conclusion, it is suggested that, through interaction with mucosal mast cells, IgE-antigen-activated myenteric neurons play a pathological role in further exacerbating the pathology of food allergy.

scholarly journals Membrane-associated phase separation: organization and function emerge from a two-dimensional milieu

2021 ◽  
Author(s):  
Jonathon A Ditlev
Keyword(s):  
Phase Separation ◽  
Cell Biology ◽  
Cellular Environment ◽  
Condensate Formation ◽  
Associated Proteins ◽  
Available Technology ◽  
And Function ◽  
Surrounding Membrane

Abstract Liquid‒liquid phase separation (LLPS) of biomolecules has emerged as an important mechanism that contributes to cellular organization. Phase separated biomolecular condensates, or membrane-less organelles, are compartments composed of specific biomolecules without a surrounding membrane in the nucleus and cytoplasm. LLPS also occurs at membranes, where both lipids and membrane-associated proteins can de-mix to form phase separated compartments. Investigation of these membrane-associated condensates using in vitro biochemical reconstitution and cell biology has provided key insights into the role of phase separation in membrane domain formation and function. However, these studies have generally been limited by available technology to study LLPS on model membranes and the complex cellular environment that regulates condensate formation, composition, and function. Here, I briefly review our current understanding of membrane-associated condensates, establish why LLPS can be advantageous for certain membrane-associated condensates, and offer a perspective for how these condensates may be studied in the future.

Mesencephalic GABA neuronal development: no more on the other side of oblivion

2014 ◽  
Vol 5 (5) ◽  
pp. 371-382 ◽  
Author(s):  
Suyan Li ◽  
Sampada Joshee ◽  
Anju Vasudevan
Keyword(s):  
Transcriptional Control ◽  
Neuronal Development ◽  
Developmental Regulation ◽  
Molecular Characteristics ◽  
Psychiatric Illnesses ◽  
Neuronal Populations ◽  
Gaba Neurons ◽  
Recent Developments ◽  
And Function ◽  
The Brain

AbstractMidbrain GABA neurons, endowed with multiple morphological, physiological and molecular characteristics as well as projection patterns are key players interacting with diverse regions of the brain and capable of modulating several aspects of behavior. The diversity of these GABA neuronal populations based on their location and function in the dorsal, medial or ventral midbrain has challenged efforts to rapidly uncover their developmental regulation. Here we review recent developments that are beginning to illuminate transcriptional control of GABA neurons in the embryonic midbrain (mesencephalon) and discuss its implications for understanding and treatment of neurological and psychiatric illnesses.

scholarly journals Isolation of Human Adrenocortical Aldosterone-Producing Cells by a Novel Immunomagnetic Beads Method

Endocrinology ◽  
2010 ◽  
Vol 151 (3) ◽  
pp. 1375-1380 ◽  
Author(s):  
Brasilina Caroccia ◽  
Ambrogio Fassina ◽  
Teresa Maria Seccia ◽  
Chiara Recarti ◽  
Lucia Petrelli ◽  
...  
Keyword(s):  
Magnetic Beads ◽  
Zona Glomerulosa ◽  
Multiple Features ◽  
Pheochromocytoma Cells ◽  
Expression Studies ◽  
Neural Cell Adhesion ◽  
Multiple Characteristics ◽  
Normal Human ◽  
Aldosterone Producing Adenoma ◽  
Gene Expression Studies

We detected intense CD56 immunostaining in the zona glomerulosa (ZG) and medulla of the normal human adrenal gland and therefore identified CD56, the neural cell adhesion molecule, as a membrane antigen specific for the ZG, aldosterone-producing adenoma (APA), and chromaffin cells. The APA and pheochromocytoma cells, which are histogenetically derived from the ZG and medulla, respectively, also showed intense CD56 immunostaining. Based on these findings we developed a strategy for isolating cells from the ZG and APA using CD56 immunobinding to magnetic beads. Morphology, gene expression studies, and aldosterone measurement confirmed that CD56 positive (+) cells were ZG and APA cells. Analysis of CD56+ cells under light and phase contrast microscopy evidenced that these cells formed clumps, as the ZG cells usually do; with electron microscopy they showed multiple features typical of a steroidogenic phenotype. Expression levels of the CD56 and the aldosterone synthase (CYP11B2) gene were markedly higher in CD56+ cells than CD56− cells (+1600 and +2100% increase, respectively). Moreover, aldosterone secretion was higher (+1380%) from CD56+ cells than from CD56− cells. Hence, this novel methodology allows isolation of a pure population of ZG and APA cells exhibiting multiple characteristics of the aldosterone-producing cells.

Expression and function of fibronectin binding integrins on rat mast cells

1995 ◽  
Vol 7 (2) ◽  
pp. 251-258 ◽  
Author(s):  
Masahiko Yasuda ◽  
Yukihito Hasunuma ◽  
Hiroyasu Adachi ◽  
Chiyoko Sekine ◽  
Tamami Sakanishi ◽  
...  
Keyword(s):  
Mast Cells ◽  
Fibronectin Binding ◽  
And Function ◽  
Rat Mast Cells

scholarly journals Emerging strategies to target RAS signaling in human cancer therapy

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Kun Chen ◽  
Yalei Zhang ◽  
Ling Qian ◽  
Peng Wang
Keyword(s):  
Human Cancer ◽  
Ras Signaling ◽  
Targeting Therapy ◽  
Ras Protein ◽  
Patients With Cancer ◽  
Mutational Status ◽  
Poor Differentiation ◽  
And Function ◽  
Nsclc Patients ◽  
Shed Light

AbstractRAS mutations (HRAS, NRAS, and KRAS) are among the most common oncogenes, and around 19% of patients with cancer harbor RAS mutations. Cells harboring RAS mutations tend to undergo malignant transformation and exhibit malignant phenotypes. The mutational status of RAS correlates with the clinicopathological features of patients, such as mucinous type and poor differentiation, as well as response to anti-EGFR therapies in certain types of human cancers. Although RAS protein had been considered as a potential target for tumors with RAS mutations, it was once referred to as a undruggable target due to the consecutive failure in the discovery of RAS protein inhibitors. However, recent studies on the structure, signaling, and function of RAS have shed light on the development of RAS-targeting drugs, especially with the approval of Lumakras (sotorasib, AMG510) in treatment of KRASG12C-mutant NSCLC patients. Therefore, here we fully review RAS mutations in human cancer and especially focus on emerging strategies that have been recently developed for RAS-targeting therapy.

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