Corticotrophin-releasing activity of desmopressin in Cushing's disease: lack of correlation between in vivo and in vitro responsiveness

Desmopressin (DDAVP), an arginine vasopressin analogue, markedly stimulates ACTH secretion in patients with Cushing's disease, in contrast to its minimal effect in normal subjects. However, little is known about the mechanisms underlying this action and it appeared to be of interest to evaluate the effect of DDAVP on ACTH-secreting pituitary adenomas in vitro, in comparison with its effect in the same patients in vivo. Pituitary adenomas from 14 patients with Cushing's disease were incubated with DDAVP, corticotrophin-releasing hormone (CRH) and DDAVP together with vasopressin receptor antagonists or CRH. Incubation with DDAVP induced a modest dose-dependent increase in ACTH concentrations which appeared maximal at 10 nM. CRH stimulated ACTH to a greater extent compared with DDAVP and potentiated the effect of DDAVP alone. The DDAVP-induced ACTH increase appeared blunted by vasopressin V(2) and V(3) receptor antagonists. V(3) receptor gene expression was detected by RT-PCR in all adenoma samples except for two which were not responsive to DDAVP in vitro but responsive to the peptide in vivo. Surprisingly, no difference in the in vitro ACTH secretory response was observed between in vivo DDAVP-responsive (ACTH peak>150% baseline) and -unresponsive (ACTH peak<120% baseline) patients, suggesting that the pituitary adenoma is not the sole mediator of the ACTH-releasing effect of DDAVP. In conclusion, the marked stimulatory effect of DDAVP observed in patients with Cushing's disease appears to be mainly dependent on an extrapituitary action, possibly the inhibition of a corticotrophin release-inhibitory factor.

Download Full-text

Decoding the genetic basis of Cushing's disease: USP8 in the spotlight

Acta Endocrinologica ◽

10.1530/eje-15-0320 ◽

2015 ◽

Vol 173 (4) ◽

pp. M73-M83 ◽

Cited By ~ 27

Author(s):

Marily Theodoropoulou ◽

Martin Reincke ◽

Martin Fassnacht ◽

Masayuki Komada

Keyword(s):

Cushing’S Disease ◽

Growth Factor Receptor ◽

Pituitary Tumors ◽

Cushing's Disease ◽

Tumor Subtypes ◽

Important Regulator ◽

Epidermal Growth ◽

Acth Secreting

Cushing's disease (CD) arises from pituitary-dependent glucocorticoid excess due to an ACTH-secreting corticotroph tumor. Genetic hits in oncogenes and tumor suppressor genes that afflict other pituitary tumor subtypes are not found in corticotrophinomas. Recently, a somatic mutational hotspot was found in up to half of corticotrophinomas in the USP8 gene that encodes a protein that impairs the downregulation of the epidermal growth factor receptor (EGFR) and enables its constitutive signaling. EGF is an important regulator of corticotroph function and its receptor is highly expressed in Cushing's pituitary tumors, where it leads to increased ACTH synthesis in vitro and in vivo. The mutational hotspot found in corticotrophinomas hyper-activates USP8, enabling it to rescue EGFR from lysosomal degradation and ensure its stimulatory signaling. This review presents new developments in the study of the genetics of CD and focuses on the USP8-EGFR system as trigger and target of corticotroph tumorigenesis.

Download Full-text

Octreotide exerts different effects in vivo and in vitro in Cushing's disease

Acta Endocrinologica ◽

10.1530/eje.0.1300125 ◽

1994 ◽

Vol 130 (2) ◽

pp. 125-131 ◽

Cited By ~ 60

Author(s):

Günter K Stalla ◽

Steffi J Brockmeier ◽

Ulrich Renner ◽

Chris Newton ◽

Michael Buchfelder ◽

...

Keyword(s):

Cell Cultures ◽

Cushing’S Disease ◽

Cushing's Disease ◽

Dependent Manner ◽

Acth Secretion ◽

Adenoma Cell ◽

Cortisol Levels ◽

Corticotropic Adenoma

Stalla GK, Brockmeier SJ, Renner U, Newton C, Buchfelder M, Stalla J, Müller OA. Octreotide exerts different effects in vivo and in vitro in Cushing's disease. Eur J Endocrinol 1994;130:125–31. ISSN 0804–4643 The effect of the long-acting somatostatin analog octreotide (SMS 201-995) on adrenocorticotropin (ACTH) secretion was studied in five patients with untreated Cushing's disease in vivo and in six human corticotropic adenoma cell cultures in vitro. For the in vivo study, 100 μg of octreotide sc was given 30 and 180 min after cannulation of the cubital vein and 100 μg of corticotropin-releasing hormone (CRH) was injected iv at 210 min. Serum ACTH and cortisol levels were measured for 390 min. In vivo, octreotide had no significant effect either on basal or CRH-stimulated ACTH levels and did not influence cortisol levels. The in vitro studies were conducted with corticotropic adenoma cell cultures derived from adenoma tissue obtained from six patients with Cushing's disease. In four of six cell cultures, octreotide (1 nmol/l–1 μmol/l) inhibited basal ACTH secretion in a dose-dependent manner. The inhibition ranged from 70 to 92% for 1 nmol/l octreotide to 14–46% for 1 μmol/l octreotide as compared to controls (100%). In three of three octreotide-responsive adenoma cell cultures investigated, CRH-stimulated ACTH secretion was suppressed by octreotide. Hydrocortisone pretreatment in vitro abolished the inhibitory effect of octreotide on ACTH secretion in one octreotide-responsive corticotropic adenoma cell culture. In conclusion, we showed that octreotide in most cases could inhibit the ACTH release from human corticotropic adenoma cells in vitro but had no suppressive effect on ACTH levels of patients with Cushing's disease in vivo. This discrepancy could be due to a somatostatin receptor down-regulation by cortisol at the hypercortisolemic state in vivo. Günter K Stalla, Max-Planck-Institute of Psychiatry, Clinical Institute, Kraepelinstr. 10, D-80804 Munich, Germany

Download Full-text

Mutation and Expression Analysis of Corticotropin-Releasing Factor 1 Receptor in Adrenocorticotropin-Secreting Pituitary Adenomas1

The Journal of Clinical Endocrinology & Metabolism ◽

10.1210/jcem.83.9.5114 ◽

1998 ◽

Vol 83 (9) ◽

pp. 3327-3331

Author(s):

K. D. Dieterich ◽

E. D. Gundelfinger ◽

D. K. Lüdecke ◽

H. Lehnert

Keyword(s):

Expression Analysis ◽

Cushing’S Disease ◽

Pituitary Adenomas ◽

Messenger Rna ◽

Corticotropin Releasing Factor ◽

Cushing's Disease ◽

Receptor Regulation ◽

R Gene ◽

Acth Secreting

The present study was designed to investigate a possible role of CRF1 receptors (CRF1-R) in the pathogenesis of Cushing’s disease. ACTH-secreting pituitary adenomas and nonsecreting pituitary adenomas have been analyzed for mutations in the CRF1-R gene by PCR and sequencing and been compared with the sequences of normal anterior pituitaries. No mutations affecting the CRF1-R protein have been found in all tumors analyzed. However, we found a significant overexpression of the CRF1-R messenger RNA in ACTH-secreting pituitary adenomas vs. inactive adenomas and normal pituitaries. We conclude that mutations of the CRF1-R are unlikely to be involved in Cushing’s disease. We suggest that the overexpression of the CRF1-R messenger RNA may be related to a disturbed receptor regulation in ACTH-secreting pituitary adenomas.

Download Full-text

Two Distinctive POMC Promoters Modify Gene Expression in Cushing’s Disease

The Journal of Clinical Endocrinology & Metabolism ◽

10.1210/clinem/dgab387 ◽

2021 ◽

Author(s):

Takako Araki ◽

Yukiko Tone ◽

Masaaki Yamamoto ◽

Hiraku Kameda ◽

Anat Ben-Shlomo ◽

...

Keyword(s):

Cushing’S Disease ◽

Methylation Status ◽

Regulatory Region ◽

Pituitary Tumors ◽

Cushing's Disease ◽

Human Pituitary ◽

Ectopic Acth ◽

Pomc Gene ◽

Acth Secreting

Abstract Context Mechanisms underlying pituitary corticotroph adenoma ACTH production are poorly understood, yet circulating ACTH levels closely correlate with adenoma phenotype and clinical outcomes. Objective We characterized the 5’ ends of proopiomelanocortin (POMC) gene transcripts, which encode the precursor polypeptide for ACTH, in order to investigate additional regulatory mechanisms of POMC gene transcription and ACTH production. Methods We examined 11 normal human pituitary tissues, 32 ACTH-secreting tumors, as well as 6 silent pituitary corticotroph adenomas (SCA) that immunostain for but do not secrete ACTH. Results We identified a novel regulatory region located near the intron2/exon3 junction in the human POMC gene, which functions as a second promoter and an enhancer. In vitro experiments demonstrated that CREB binds the second promoter and regulates its transcriptional activity. The second promoter is highly methylated in SCA, partially demethylated in normal pituitary tissue, and highly demethylated in pituitary and ectopic ACTH-secreting tumors. In contrast, the first promoter is demethylated in all POMC-expressing cells and is highly demethylated only in pituitary ACTH-secreting tumors harboring the USP8 mutation. Demethylation patterns of the second promoter correlate with clinical phenotypes of Cushing’s disease. Conclusion We identified a second POMC promoter regulated by methylation status in ACTH-secreting pituitary tumors. Our findings open new avenues for elucidating subcellular regulation of the hypothalamic-pituitary-adrenal axis and suggest the second POMC promoter may be a target for therapeutic intervention to suppress excess ACTH production.

Download Full-text

Cushing's disease and hypertension: in vivo and in vitro study of the role of the renin-angiotensin-aldosterone system and effects of medical therapy

Acta Endocrinologica ◽

10.1530/eje-13-0477 ◽

2014 ◽

Vol 170 (2) ◽

pp. 181-191 ◽

Cited By ~ 9

Author(s):

R van der Pas ◽

J H M van Esch ◽

C de Bruin ◽

A H J Danser ◽

A M Pereira ◽

...

Keyword(s):

Blood Pressure ◽

Cushing’S Disease ◽

In Vitro Study ◽

Cushing's Disease ◽

Ang Ii ◽

Renin Angiotensin Aldosterone System ◽

Renin Angiotensin ◽

Raas Blockade

Objective/methodsCushing's disease (CD) is often accompanied by hypertension. CD can be treated surgically and, given the expression of somatostatin subtype 5 and dopamine 2 receptors by corticotroph pituitary adenomas, pharmacologically. Indeed, we recently observed that stepwise medical combination therapy with the somatostatin-analog pasireotide, the dopamine-agonist cabergoline, and ketoconazole (which directly suppresses steroidogenesis) biochemically controlled CD patients and lowered their blood pressure after 80 days. Glucocorticoids (GC) modulate the renin–angiotensin–aldosterone system (RAAS) among others by increasing hepatic angiotensinogen expression and stimulating mineralocorticoid receptors (MR). This study therefore evaluated plasma RAAS components in CD patients before and after drug therapy. In addition, we studied whether cabergoline/pasireotide have direct relaxant effects in angiotensin II (Ang II)-constricted iliac arteries of spontaneously hypertensive rats, with and without concomitant GR/MR stimulation with dexamethasone or hydrocortisone.ResultsBaseline concentrations of angiotensinogen were elevated, while renin and aldosterone were low and suppressed, respectively, even in patients treated with RAAS-blockers. This pattern did not change after 80 days of treatment, despite blood pressure normalization, nor after 4 years of remission. In the presence of dexamethasone, pasireotide inhibited Ang II-mediated vasoconstriction.ConclusionsThe low plasma renin concentrations, even under RAAS blockade, in CD may be the consequence of increased GC-mediated MR stimulation and/or the elevated angiotensinogen levels in such patients. The lack of change in RAAS-parameters despite blood pressure and cortisol normalization suggests persisting consequences of long-term exposure to cortisol excess. Finally, pasireotide may have a direct vasodilating effect contributing to blood pressure lowering.

Download Full-text

Pituitary Adenomas with Changing Phenotype: A Systematic Review

Experimental and Clinical Endocrinology & Diabetes ◽

10.1055/a-1120-8277 ◽

2020 ◽

Vol 128 (12) ◽

pp. 835-844

Author(s):

Fernando Guerrero-Pérez ◽

Agustina Pia Marengo ◽

Noemi Vidal ◽

Carles Villabona

Keyword(s):

Systematic Review ◽

Cushing’S Disease ◽

Pituitary Adenomas ◽

Tumor Resection ◽

Cushing's Disease ◽

Complete Tumor Resection ◽

Null Cell ◽

Acth Secreting ◽

Complete Tumor

Abstract Purpose and Methods Phenotype transformation in pituitary adenomas (PA) is a little known and unexpected clinical phenomenon. We describe two illustrative cases and performed a systematic review of cases reported in literature. Results Case 1: A 24-year-old woman underwent surgery because of Cushing’s disease. A complete tumor resection and hypercortisolism resolution was achieved. Two years later, tumor recurred but clinical and hormonal hypercortisolism were absent. Case 2: A 77-year-old woman underwent surgery due to acromegaly. A complete tumor resection and GH excess remission was achieved. Four years later, tumor recurred but clinical and hormonal acromegaly was ruled out. Search of literature: From 20 patients (including our cases), 75% were female with median age 45 (19) years. Ten patients (50%) had initially functioning PA: 8 switched to NFPA (5 ACTH-secreting PA, 2 prolactinomas and 1 acromegaly) and 2 exchanged to acromegaly from TSH-secreting PA and microprolactinoma. One patient developed a pituitary carcinoma from ACTH-secreting PA. Ten patients (50%) initially had NFPA; 9 developed Cushing’s disease (4 silent corticotroph adenomas, 4 null cell PA and 1 managed conservatively). One patient with silent somatotroph PA changed to acromegaly. Treatments before transformation were surgery (80%), radiotherapy (40%), pharmacological (40%) and in 2 patients switching happened without any treatment. Median follow-up until transformation was 72 months (range 12–276). Conclusion PA can change from functioning to (NF) non-functioning (vice versa) and even exchange their hormonal expression. Clinicians should be aware and a careful lifelong follow-up is mandatory to detect it.

Download Full-text

TMOD-49. ANTITUMOR EFFICACY OF ANTI-PDL1 IN ACTH-SECRETING PITUITARY ADENOMAS: AN IMMUNOTHERAPEUTIC APPROACH FOR CUSHING’S DISEASE

Neuro-Oncology ◽

10.1093/neuonc/nox168.1085 ◽

2017 ◽

Vol 19 (suppl_6) ◽

pp. vi264-vi265

Author(s):

Aladine A Elsamadicy ◽

S Harrison Farber ◽

Pakawat Chongsathidkiet ◽

Hanna Kemeny ◽

Karolina Woroniecka ◽

...

Keyword(s):

Cushing’S Disease ◽

Pituitary Adenomas ◽

Antitumor Efficacy ◽

Cushing's Disease ◽

Immunotherapeutic Approach ◽

Acth Secreting

Download Full-text

In Vitro Secretion of Adenoma and Anterior Lobe Cells in Two Typical Cases of Cushing's Disease

Neurosurgery ◽

10.1227/00006123-198305000-00012 ◽

1983 ◽

Vol 12 (5) ◽

pp. 549-554 ◽

Cited By ~ 6

Author(s):

Dieter K. Lüdecke ◽

Martin Schabet ◽

Wolfgang Saeger

Keyword(s):

Cushing’S Disease ◽

Anterior Lobe ◽

Cushing's Disease ◽

Acth Secretion ◽

Acth Cell ◽

Superfusion System ◽

Adenomatous Tissue ◽

Corticoid Feedback ◽

Acth Secreting

Abstract Fragments of adrenocorticotropic hormone (ACTH) cell adenomas and anterior lobes of two patients with Cushing's disease were obtained by transnasal operation. Both patients showed the typical clinical course, with postoperative ACTH deficit and all other pituitary functions intact. Equivalent specimens of tissue were investigated by immunocytology and in a superfusion system. The majority of adenoma cells were ACTH-positive, whereas ACTH-secreting cells of the anterior lobes were mostly inactive and were reduced in number. In vitro, adenomatous tissue showed high ACTH secretion into the superfusion medium, which was increased significantly after vasopressin application. Corticoid feedback was impaired. Anterior lobe cells exhibited a significant spontaneous ACTH secretion that was reduced by cortisol, but not stimulated by vasopressin. These results support the concept of an impaired corticoid feedback at the adenoma level in the presence of suppressed ACTH secretion of the para-adenomatous anterior lobe.

Download Full-text

Multihormonal response to corticotropin-releasing hormone in inferior petrosal sinus blood of one patient with Cushing's disease: comparison with in vitro secretion of the tumoral corticotropes

Acta Endocrinologica ◽

10.1530/acta.0.1270284 ◽

1992 ◽

Vol 127 (3) ◽

pp. 284-288 ◽

Cited By ~ 7

Author(s):

Antoine Tabarin ◽

Jean-Benoît Corcuff ◽

Michel Rashedi ◽

Reine Angibeau ◽

Jean-Marie Caille ◽

...

Keyword(s):

Cushing’S Disease ◽

Cushing's Disease ◽

Pituitary Cells ◽

Inferior Petrosal Sinus ◽

Inferior Petrosal Sinus Sampling ◽

Tsh Secretion ◽

Corticotropic Adenoma ◽

Stimulation Of

A multihormonal response to CRH during inferior petrosal sinus sampling in patients with Cushing's disease has recently been described. Whether it reflects multihormonal secretion by the corticotropic adenoma, or secretion by non-tumorous adjacent cells via paracrine mechanisms remains debatable. We have compared the effect of CRH on ACTH, GH, PRL and TSH secretion during inferior petrosal sinus sampling with its effect on the in vitro secretion of the corticotropic adenoma after excision in one case of Cushing's disease. Before CRH injection in vivo results show significant central-peripheral gradients for all hormones but only ACTH lateralized to the side of the tumor. After CRH administration, the petrosal concentrations of all hormones increased preferentially on the side of the adenoma resulting in significant intersinus gradients: 8.1 for ACTH, 2.0 for GH, 1.8 for PRL and 1.5 for TSH. In vitro results: the adenoma cells were immunostainable for ACTH only. In culture, they secreted ACTH only. Addition of CRH to the culture induced a mean increase of 160% in ACTH secretion but GH, PRL and TSH remained undetectable. Our results favor the hypothesis that the multihormonal response to CRH seen during inferior petrosal sinus sampling in Cushing's disease reflects a paracrine stimulation of the adjacent non-tumorous pituitary cells by the corticotropic adenoma.

Download Full-text

Sexual Dimorphism in Cellular and Molecular Features in Human ACTH-Secreting Pituitary Adenomas

Cancers ◽

10.3390/cancers12030669 ◽

2020 ◽

Vol 12 (3) ◽

pp. 669 ◽

Cited By ~ 2

Author(s):

Francesca Pecori Giraldi ◽

Maria Francesca Cassarino ◽

Antonella Sesta ◽

Mariarosa Terreni ◽

Giovanni Lasio ◽

...

Keyword(s):

Sexual Dimorphism ◽

Differentially Expressed Genes ◽

Cushing’S Disease ◽

Pituitary Adenomas ◽

Differentially Expressed ◽

Cushing's Disease ◽

Female Patients ◽

Molecular Features ◽

Male Patients ◽

Acth Secreting

(1) Background. Cushing’s disease presents gender disparities in prevalence and clinical course. Little is known, however, about sexual dimorphism at the level of the corticotrope adenoma itself. The aim of the present study was to evaluate molecular features of ACTH-secreting pituitary adenomas collected from female and male patients with Cushing’s disease. (2) Methods. We analyzed 153 ACTH-secreting adenomas collected from 31 men and 122 women. Adenomas were established in culture and ACTH synthesis and secretion assessed in basal conditions as well as during incubation with CRH or dexamethasone. Concurrently, microarray analysis was performed on formalin-fixed specimens and differences in the expression profiles between specimens from male and female patients identified. (3) Results. ACTH medium concentrations in adenomas obtained from male patients were significantly lower than those observed in adenomas from female patients. This could be observed for baseline as well as modulated secretion. Analysis of corticotrope transcriptomes revealed considerable similarities with few, selected differences in functional annotations. Differentially expressed genes comprised genes with known sexual dimorphism, genes involved in tumour development and genes relevant to pituitary pathophysiology. (4) Conclusions. Our study shows for the first time that human corticotrope adenomas present sexual dimorphism and underlines the need for a gender-dependent analysis of these tumours. Differentially expressed genes may represent the basis for gender-tailored target therapy.

Download Full-text