Pathogenesis Of Acromegaly Research Articles

MicroRNA in Acromegaly: Involvement in the Pathogenesis and in the Response to First-Generation Somatostatin Receptor Ligands.

Acromegaly is a chronic and systemic disease due to excessive growth hormone and insulin-like growth factor type I caused, in the vast majority of cases, by a GH-secreting pituitary adenoma. About 40% of these tumors have somatic mutations in the stimulatory G protein alpha-subunit 1 gene. The pathogenesis of the remaining tumors, however, is still not fully comprehended. Surgery is the first-line therapy for these tumors, and first-generation somatostatin receptor ligands (fg-SRL) are the most prescribed medications in patients who are not cured by surgery. MicroRNAs are small, non-coding RNAs that control the translation of many mRNAs, and are involved in the post-transcriptional regulation of gene expression. Differentially expressed miRNAs can explain differences in the pathogenesis of acromegaly and tumor resistance. In this review, we focus on the most validated miRNAs, which are mainly involved in acromegaly’s tumorigenesis and fg-SRL resistance, as well as in circulating miRNAs in acromegaly.

Germline or somatic GPR101 duplication leads to X-linked acrogigantism: a clinico-pathological and genetic study.

Non-syndromic pituitary gigantism can result from AIP mutations or the recently identified Xq26.3 microduplication causing X-linked acrogigantism (XLAG). Within Xq26.3, GPR101 is believed to be the causative gene, and the c.924G > C (p.E308D) variant in this orphan G protein-coupled receptor has been suggested to play a role in the pathogenesis of acromegaly.We studied 153 patients (58 females and 95 males) with pituitary gigantism. AIP mutation-negative cases were screened for GPR101 duplication through copy number variation droplet digital PCR and high-density aCGH. The genetic, clinical and histopathological features of XLAG patients were studied in detail. 395 peripheral blood and 193 pituitary tumor DNA samples from acromegaly patients were tested for GPR101 variants.We identified 12 patients (10 females and 2 males; 7.8 %) with XLAG. In one subject, the duplicated region only contained GPR101, but not the other three genes in found to be duplicated in the previously reported patients, defining a new smallest region of overlap of duplications. While females presented with germline mutations, the two male patients harbored the mutation in a mosaic state. Nine patients had pituitary adenomas, while three had hyperplasia. The comparison of the features of XLAG, AIP-positive and GPR101&AIP-negative patients revealed significant differences in sex distribution, age at onset, height, prolactin co-secretion and histological features. The pathological features of XLAG-related adenomas were remarkably similar. These tumors had a sinusoidal and lobular architecture. Sparsely and densely granulated somatotrophs were admixed with lactotrophs; follicle-like structures and calcifications were commonly observed. Patients with sporadic of familial acromegaly did not have an increased prevalence of the c.924G > C (p.E308D) GPR101 variant compared to public databases.In conclusion, XLAG can result from germline or somatic duplication of GPR101. Duplication of GPR101 alone is sufficient for the development of XLAG, implicating it as the causative gene within the Xq26.3 region. The pathological features of XLAG-associated pituitary adenomas are typical and, together with the clinical phenotype, should prompt genetic testing.Electronic supplementary materialThe online version of this article (doi:10.1186/s40478-016-0328-1) contains supplementary material, which is available to authorized users.

The Glucose-Dependent Insulinotropic Polypeptide Receptor is Overexpressed Amongst GNAS1 Mutation-Negative Somatotropinomas and Drives Growth Hormone (GH)-Promoter Activity in GH3 Cells

Somatic mutations in the GNAS1 gene, encoding the α-subunit of the heterotrimeric stimulatory G protein (Gαs), occur in approximately 40% of growth hormone (GH)-secreting pituitary tumours. By altering the adenylate cyclase-cAMP-protein kinase A pathway, they unequivocally give somatotroph cells a growth advantage. Hence, the pathogenesis of somatotropinomas could be linked to anomalies in receptors coupled to the cAMP second-messenger cascade. Among them, the glucose-dependent insulinotropic polypeptide receptor (GIPR) is already known to play a primary role in the impaired cAMP-dependent cortisol secretion in patients affected by food-dependent Cushing's syndrome. In the present study, 43 somatotropinomas and 12 normal pituitary glands were investigated for GIPR expression by quantitative reverse transcriptase-polymerase chain reaction, western blotting and immunohistochemistry. Tumoural specimens were also evaluated for GNAS1 mutational status. The effect of GIPR overexpression on cAMP levels and GH transcription was evaluated in an in vitro model of somatotropinomas, the GH-secreting pituitary cell line GH3. GIPR was expressed at higher levels compared to normal pituitaries in 13 GNAS1 mutation-negative somatotropinomas. GIP stimulated adenylyl cyclase and GH-promoter activity in GIPR-transfected GH3 cells, confirming a correct coupling of GIPR to Gαs. In a proportion of acromegalic patients, GIPR overexpression appeared to be associated with a paradoxical increase in GH after an oral glucose tolerance test. Whether GIPR overexpression in acromegalic patients may be associated with this paradoxical response or more generally involved in the pathogenesis of acromegaly, as suggested by the mutually exclusive high GIPR levels and GNAS1 mutations, remains an open question.

Molecular biology of the somatotroph adenoma cell

Despite considerable progress during the last 15 years, the pathogenesis of acromegaly is still poorly understood. GH producing adenoma cells are monoclonal in origin and as many other neoplastic cell types, are most likely the result of a series of genetic alterations. This involves expression of oncogenes such as gsp, which was reported to be expressed in 30 to 40% of somatotroph adenmomas and which induces constitutive GH production. Loss of heterozygosity and subsequent loss or inactivation of different tumour suppressors has also been found in sporadic and hereditary somatotroph tumours. However, none of the oncogenes or tumour suppressors studied so far has been consistently overexpressed or lost in all somatotroph adenomas demonstrating the heterogeneity of this tumour type. Once a transformed somatotroph tumour cells has emerged, the developing tumour slowly, but continuously expands and starts to produce excessive amounts of GH. This indicates that the mechanisms that inhibit GH secretion (feedback-inhibition through IGF-1; GH suppression by hypothalamic somatostatin) are impaired. Desensitation, downregulation or mutation of the IGF-1 receptor or the somatostatin receptor subtypes as well as disturbances of signalling induced by these receptors have been discussed as the cause of missing inhibitory control of GH release. Moreover, impaired auto-/paracrine-acting intrapituitary growth factor and cytokine loops seem to be disturbed and may contribute to the pathophysiology and progression of somatotroph tumors. In the presentation intra- and intercellular molecular changes associated with somatotroph pituitary adenomas will be summarised and discussed.

Elevated intraocular levels of insulin-like growth factor I in a diabetic patient with acromegaly

Growth factors have become increasingly associated with various events of vitreoretinal disease. In the presence of proliferative retinopathy, elevated levels of insulin growth factor I (IGF-I) have been demonstrated in the vitreous. IGF-I expression itself is regulated by growth hormone, a hormone that is involved in the pathogenesis of acromegaly. In the present paper we report on the detection of very high IGF-levels in the subretinal fluid of a patient with acromegaly who was operated for retinal detachment. With a very sensitive RIA assay we found a four-fold elevation of the serum levels of IGF-I (604 ng/ml) and a high level of IGF-1 in the subretinal fluid (13 ng/ml). It is concluded that acromegaly may lead to an overexpression of IGF-I not only seen as increased concentrations in serum and vitreous, but in the subretinal space as well. This study emphasizes the need for further investigation of growth factor presence in pathological cavities, such as the subretinal space during retinal detachments.

Imprinting of the G(s)alpha gene GNAS1 in the pathogenesis of acromegaly.

Approximately 40% of growth hormone-secreting pituitary adenomas have somatic mutations in the GNAS1 gene (the so-called gsp oncogene). These mutations at codon 201 or codon 227 constitutively activate the alpha subunit of the adenylate cyclase-stimulating G protein G(s). GNAS1 is subject to a complex pattern of genomic imprinting, its various promoters directing the production of maternally, paternally, and biallelically derived gene products. Transcripts encoding G(s)alpha are biallelically derived in most human tissues. Despite this, we show here that in 21 out of 22 gsp-positive somatotroph adenomas, the mutation had occurred on the maternal allele. To investigate the reason for this allelic bias, we also analyzed GNAS1 imprinting in the normal adult pituitary and found that G(s)alpha is monoallelically expressed from the maternal allele in this tissue. We further show that this monoallelic expression of G(s)alpha is frequently relaxed in somatotroph tumors, both in those that have gsp mutations and in those that do not. These findings imply a possible role for loss of G(s)alpha imprinting during pituitary somatotroph tumorigenesis and also suggest that G(s)alpha imprinting is regulated separately from that of the other GNAS1 products, NESP55 and XLalphas, imprinting of which is retained in these tumors.

Absence of somatostatin receptor type 2 A mutations and gip oncogene in pituitary somatotroph adenomas.

Somatostatin, acting via specific receptors in the anterior pituitary, tonically inhibits pituitary growth hormone secretion and somatotroph proliferation. Reduction of growth hormone secretion and tumour regression in GH-secreting pituitary adenomas treated with long-acting somatostatin analogues varies widely. In 30-40% of these tumours dominant somatic mutations of the Gsalpha gene (gsp) have been demonstrated leading to constitutive adenylyl cyclase induction. A relationship between somatostatin sensitivity and tumour pathogenesis in some tumours has been suggested. Changes in the function of the somatostatin receptor or intracellular signal elements may be of relevance. Somatostatin receptor type 2 A (sst2A) and Gi2 are proposed to mediate selectively the inhibition of GH release in the somatotroph. We therefore investigated the presence of sst2A mutations and gip oncogene in somatotrophic pituitary adenomas. Tumour samples from 15 patients with pituitary somatotroph adenomas were obtained. RNA was isolated and used for reverse transcription and subsequent polymerase chain reaction. All samples were screened for the presence of sst2A mutations and of the gip oncogene by SSCP analysis and sequencing. For comparison, the gsp oncogene was examined. The relationship between clinical data and molecular analysis results was investigated. Seven of the tumours harboured a gsp mutation. No mutations affecting the sst2A protein were found in any of the tumours analysed. Furthermore, gip oncogene was absent in all tumours. Mutations of the somatostatin receptor type 2 A and the gip oncogene are unlikely to be involved in the pathogenesis of acromegaly.

Molecular pathogenesis of acromegaly.

Acromegaly is generally caused by growth hormone (GH) hypersecretion from a benign, monoclonal pituitary adenoma. As in other neoplastic conditions, pituitary tumor formation and dysregulated hormone secretion are most likely the ultimate result of a series of genetic alterations. A number of molecular and biochemical defects have been associated with pituitary tumorigenesis. Molecular events such as tumor suppressor gene inactivation and oncogene activation involved in pituitary tumor progression are examined. The role of hypothalamic regulatory hormones and hereditary syndromes involving acromegaly are also discussed.

The IGF-I receptor sub-membrane domain is intact in GH-secreting pituitary tumours.

Clinical acromegaly is characterized by dysregulation of somatotroph GH secretion in the presence of high circulating serum IGF-I levels. Physiologically, IGF-I exerts a negative feedback on GH secretion at both the hypothalamic and the pituitary levels. We have previously shown that the 943 and 950 tyrosine residues in the IGF-I receptor beta-subunit are required for ligand signalling to the GH gene, as substitution of these residues abrogates IGF-I signal transduction. To determine whether a mutation within the IGF-I receptor submembrane domain may be involved in the pathogenesis of GH secreting tumours, we studied this region in these tumours. Exon 15 of the IGF-I receptor containing both the 943 and 950 tyrosines was analysed in 19 GH-secreting tumours by single-strand conformation polymorphism (SSCP) analysis of polymerase chain reaction (PCR) products. Tumour DNA and patients' lymphocyte DNA, which served as normal controls, were analysed. All samples exhibited normal migration patterns in the SSCP analysis which was further confirmed by direct DNA sequencing. We conclude that mutations in the IGF-I receptor sub-membrane domain which disrupt the negative feedback loop are not involved in the pathogenesis of acromegaly.

Molecular screening of pituitary adenomas for gene mutations and rearrangements.

Although pituitary tumors arise as benign monoclonal neoplasms, genetic alterations have not readily been identified in these adenomas. We studied restriction fragment abnormalities involving the GH gene locus, and mutations in the p53 and H-, K- and N-ras genes in 22 human GH cell adenomas. Twenty two nonsecretory adenomas were also examined for p53 and ras gene mutations. Seven prolactinoma DNA samples were tested for deletions in the multiple endocrine neoplasia-1 (MEN-1) locus, as well as for rearrangements in the hst gene, a member of the fibroblast growth factor family. Pituitary adenoma tissue and lymphocytes were obtained from patients at the time of transsphenoidal surgery. In DNA from GH-cell adenomas, identical GH restriction patterns were detected in both pituitary and lymphocyte DNA in all patients and in one patient with a mixed GH-TSH cell adenoma. Using polymerase chain reaction (PCR)-single stranded conformation polymorphism analysis, no mutations were detected in exons 5, 6, 7, and 8 of the p53 gene in GH cell adenomas nor in 22 nonsecretory adenomas. Codons 12/13 and 61 of H-ras, K-ras, and N-ras genes were also intact in GH cell adenomas and in nonsecretory adenomas. Site-specific probes for chromosome 11q13 including PYGM, D11S146, and INT2 were used in 7 sporadic PRL-secreting adenomas to detect deletions of the MEN-1 locus on chromosome 11. One patient was identified with a loss of 11p, and the remaining 6 patients did not demonstrate loss of heterozygosity in the pituitary 11q13 locus, compared to lymphocyte DNA. None of these patients, demonstrated hst gene rearrangements which also maps to this locus. These results show that p53 and ras gene mutations are not common events in the pathogenesis of acromegaly and nonsecretory tumors. Although hst gene rearrangements and deletions of 11q13 are not associated with sporadic PRL-cell adenoma formation, a single patient was detected with a partial loss of chromosome 11, including the putative MEN-1 site.

Impaired glucose tolerance coincides with abnormal release of growth hormone following a glucose load as well as in response to TRH in acromegaly.

It is known that some acromegalic patients exhibited a paradoxical release of growth hormone (GH) after glucose administration. We have attempted to investigate a relationship between the paradoxical GH secretion with the abnormal glucose tolerance test present in some cases of acromegaly. We also studied the inappropriate increase in GH levels following thyrotropin releasing hormone (TRH) injection which is present in some acromegalics. We found that only those patients who had an abnormal glucose tolerance test exhibited simultaneously, the paradoxical release of GH, moreover, the same patients showed GH release following TRH administration. This observation suggests that some acromegalics have an abnormality in their hypothalamic glucose receptor and such abnormality is associated with abnormal GH secretion when TRH is administered. On basis of these findings it is suggested that the hypothalamus may play an important role in the pathogenesis of acromegaly in these cases.

The pathogenesis of acromegaly. Clinical and immunocytochemical analysis in 75 patients.

A series of 75 patients with acromegaly and immunocytochemically characterized pituitary adenomas has been analyzed. Tumors secreting growth hormone (GH) only were found in 21% of cases. The remainder had tumors immunoreactive for more than one pituitary hormone: GH and prolactin in 31%; GH, prolactin, and glycoprotein in 40%; and GH and glycoprotein in 8%. Microadenomas were surgically treated in 17 patients with a success rate of 82%. Overall, normalization of basal GH secretion (to less than or equal to 5 ng/ml) was achieved in 54% of cases. The implications of these findings for the pathogenesis and neurosurgical management of acromegaly are discussed.

Pathophysiology of acromegaly.

This review discusses the pathophysiology of acromegaly. Acromegaly has been classified in this paper into distinct entities based on etiology, ultrastructural features of the pituitary, and cytogenesis. This classification has been proposed based on clinical signs, immunoperoxidase techniques, transmission electromicroscopy and immunoelectron microscopy. Pituitary causes of acromegaly include densely granulated adenomas, sparsely granulated adenomas, mixed growth hormone and prolactin cell adenomas, acidophil stem cell adenomas, mammosomatotroph cell adenomas, and pleurihormonal adenomas. GH cell hyperplasia and GH cell carcinoma are also discussed. Extrapituitary causes of acromegaly include eutopic GH cell adenoma in the sphenoid sinus or parapharyngeal region and excess GHRF secretion which may be eutopic or ectopic. The pathological, clinical, and biochemical evidence in favor of a pituitary or hypothalamic etiology of acromegaly has been reviewed. Finally, a multistage theory of GH cell tumorigenesis has been proposed as a model in an attempt to unify the genetic, environmental and biochemical factors implicated in the pathogenesis of acromegaly.

THE PATHOGENESIS OF ACROMEGALY.

THE PATHOGENESIS OF ACROMEGALY.

Pathogenesis of Acromegaly

Pathogenesis of Acromegaly