cAMP pathway and pituitary tumorigenesis

The pituitary is the target of different neurohormones that have a crucial role in the control of cell differentiation, cell proliferation and hormone secretion by recognizing specific receptors belonging to the G Protein-Coupled Receptor super-family (GPCR). Evidence from in vitro studies and naturally occurring human diseases indicate that several endocrine cells, and particularly somatotrophs, recognize cAMP as a growth factor. Accordingly, mutations of the alpha subunit of the stimulatory G protein gene (GNAS) leading to the constitutive activation of adenylyl cyclase (i.e. gsp oncogene) have been recognized in a significant proportion of GH-secreting pituitary adenomas. The role of cAMP in the control of cell proliferation in selected cell types and in particular in somatotroph cells has been further confirmed by identification of genetics defect affecting the regulatory subunit IA of PKA. The role of cAMP in the control of cell proliferation as well as the crosstalk with different intracellular signalling pathways will be discussed.     

Epigenetic change in pituitary tumorigenesis

Throughout the genome CpG dinucleotides are found at one-fifth of their expected frequency and their rarity is further marked by the fact that 70% are methylated. In contrast, CpG islands (CGI), found associated with the promoters of many genes, have maintained their expected frequency of this dinucleotide, and remain unmethylated. Inappropriate methylation of CGIs is associated with histone deacetylation and gene silencing, while methylation of CpGs outside of CGIs is associated with significantly higher mutation rates. Methylation of CGIs is a frequent event in numerous tumour types including those that arise within the pituitary gland. Several studies now show highly frequent methylation of the p16 gene that is significantly associated with loss of cognate protein and that appears to be an early change in pituitary tumorigenesis. Collectively, studies show that somatotrophinomas are an infrequent target for p16 CGI methylation. However, in this pituitary tumour subtype, loss of pRb is associated with either CGI methylation or micro-deletion within the protein-pocket binding domain. As in other tumour types loss of p16 or RB1 appear to be mutually exclusive events in non-functional adenomas and somatotrophinomas respectively. Investigation of the Death Associated Protein Kinase gene shows that loss of its protein (DAPK), a pro-apoptotic molecule, in pituitary tumours is also associated with either methylation or deletion within its associated CGI. In the case of DAPK, however, these changes segregate with invasive pituitary tumours irrespective of tumour subtype. Methylation represents a positive signal that can be detected with exquisite sensitivity; in addition, this change targets multiple genes that show tumour type specificity. Taken together, the detection of DNA methylation changes, using either a panel of predefined marker-islands, or CGI arrays, provides the opportunity to generate "methylation profiles". This new knowledge will increase our understanding of tumour biology and could ultimately aid medical management in these different tumour types, including those of pituitary origin.     

Molecular mechanisms involved in adrenocortical tumorigenesis

The adrenocortical tumorigenesis is a complex process, which involves multiple genetic changes. A better knowledge on the mechanisms involved in tumor development would enable an early identification of malignant disease and also lead to the development of new treatment strategies. Although in the recent years a large amount of data was produced, the exact mechanisms that lead to adrenocortical tumor development remains poorly understood. Most of the studies produced were based on the candidate-gene strategy, which has its own limitations. A genome-wide approach, such as microarrays, will surely shed some light into the mechanisms responsible for adrenocortical tumorigenesis. In this review, we summarize the most recent data available on this complex process.     

Molecular and trophic mechanisms of tumorigenesis.

A significant proportion of pituitary macroadenomas, and by definition all microadenomas, regain trophic stability after an initial period of deregulated growth. Classical proto-oncogene activation and tumor suppressor mutation are rarely responsible, and no histologic or molecular markers reliably predict behavior. GNAS1 activation and the mutations associated with multiple endocrine neoplasia type 1 and Carney complex, aryl hydrocarbon receptor interacting protein gene mutations, and a narrowing region of chromosome 11q13 in familial isolated acromegaly together account for such a small proportion of pituitary adenomas that the pituitary adenoma pathogenic epiphany is surely yet to come.     

Molecular and genetic defects in colorectal tumorigenesis

Colorectal cancers, whether sporadic or hereditary, are caused by a defined set of molecular events. There are at least two different pathogenetic pathways for colorectal cancer: the chromosomal instability pathway and the microsatellite instability pathway; the two major inherited syndromes, familial adenomatous polyposis (FAP) and hereditary non-polyposis colorectal cancer (HNPCC), are examples of these two mechanisms. These different pathways, however, converge on common pathological entities that have crucial functions in the regulation of normal crypt homeostasis. Preventive strategies aimed at reversing these changes, or therapeutic interventions targeting cell populations with these alterations, should be most efficacious. Genetic testing for inherited syndromes is now available and allows appropriate management of these disorders. Further insight into colorectal tumorigenesis pathways can lead to the development of useful prognostic indicators and target preventive and therapeutic strategies in the management of colorectal cancer.     

Molecular aspects of cholangiocarcinoma

Novel targets for therapeutic or chemopreventive approaches against cholangiocarcinoma (CCA) are urgently needed. In this review article, we discuss the molecular aspects of CCA including the role of erbB receptor tyrosine kinases (RTKs), downstream signaling pathways of these erbB RTKs, inflammatory mediators during gallbladder carcinogenesis and bile acids based on our study using a mouse model for human CCA (BK5.erbB2 mice) as well as additional information in the literature.     

Molecular aspects of myocarditis

Myocarditis is an acquired form inflammatory heart muscle disease, manifested as acute and chronic conditions. While many etiologies have been reported, the most common cause of this disease is infection, primarily viral. Typically, the specific causative agent(s) and mechanism(s) are elusive. Over the past several years, various new findings have added to our understanding of myocarditis. These include the identification of adenoviruses as important causative agents, a new receptor protein likely to play an important role in the virulence of certain agents affecting the myocardium, and the effect of viruses on the cardiac cytoskeleton. This report reviews the current understanding of myocarditis, proposes a hypothesis about the long-term sequelae, and suggests possible new therapeutic strategies.     

Growth hormone-releasing hormone and pituitary development, hyperplasia and tumorigenesis.

Growth hormone-releasing hormone (GHRH) is essential for expansion of the somatotrope lineage during pituitary development, and excessive GHRH secretion and/or action results in unregulated somatotrope proliferation and neoplastic transformation. Our understanding of the molecular and morphological bases for these effects from both animal and clinical studies has greatly increased during the past decade. However, many features of the cellular pathways remain to be defined, including the interaction of other genes in the multistep process of somatotrope tumorigenesis.     

Ovarian dependence for pituitary tumorigenesis in D2 dopamine receptor-deficient mice

Hypophyseotropic dopamine exerts a tonic inhibitory tone on pituitary lactotrophs by the activation of dopamine D2 receptors (D2R). Ablation of D2R through gene knock-out approaches results in hyperprolactinemia and prolactinomas. This phenotype is more severe and develops more rapidly in female mice. We tested whether the female hypersensitivity is due solely to the loss of D2R inhibitory tone or concomitant stimulation by ovarian factors. C57BL/6J congenic D2R(-/-) mice were ovariectomized at 2 months of age and serum PRL levels were measured serially. Ovariectomy attenuated hyperprolactinemia and after 18 months, D2R(-/-) mice had average pituitary weights of 4 mg, compared with 60 mg in the intact group. 17beta-Estradiol did not restore PRL secretion or pituitary weight. Although the pharmacologic dose of estradiol slightly increased pituitary weight in wild-type and D2R(-/-) mice, it inhibited serum PRL in both intact and ovariectomized females and in castrated males. For comparison, we tested the estradiol response of wild-type 129S6/SvEv mice in the same paradigm and found the expected increase in pituitary weight and serum PRL. Our results demonstrate that the development of hyperprolactinemia and prolactinomas in mice lacking D2R is dependent on ovarian stimulation and likely involves a factor(s) in addition to estrogen. Furthermore, we showed that estradiol-induced proliferation and PRL secretion can be differentially regulated in a strain-specific manner. These findings illustrate the importance of genetic background when analyzing endocrine regulation in mutant mouse models.     

Molecular genetics of pituitary tumours.

The last several years have seen a significant increase in our understanding of the molecular and biochemical changes associated with pituitary tumour initiation and progression. The combined data, from several groups, now allow a tentative map to be drawn showing that reduction to hemizygosity at several chromosomal loci (10q, 11q13 and 13q) is associated with the transition to the invasive phenotype, while loss on chromosome 9p and methylation of the tumour suppressor gene p16 appear to occur early in pituitary tumorigenesis. Changes in the expression/status of several genes and/or proteins including p53, the cAMP response element-binding factor (CREB), growth hormone-releasing hormone (GHRH), nm23, p16 and p27 have also been identified along this multi-step pathway. Prospective studies will determine whether these markers are truly predictive of subsequent tumour behaviour and can be used to aid clinical management in a manner not possible when current histological criteria are used.     

Molecular aspects of disaccharidase deficiencies

We have described the methods used for studying the biosynthesis and the post-translational processing of sucrase-isomaltase (SI), lactase-phlorizin hydrolase (LPH) and maltase-glucoamylase (MGA) in human small intestinal mucosa. Our results are discussed in the context of findings by other researchers. A surprising finding coming out of all these studies is that SI, LPH and MGA are structurally quite different. SI and LPH are both synthesized as large molecular weight precursors which are proteolytically processed to the mature enzymes. In the case of SI, this processing occurs after insertion of the precursor into the brush border membrane and is catalysed by pancreatic proteases; the mature form consists of the two subunits sucrase and isomaltase, the latter containing an N-terminal peptide anchor. Proteolytic processing of the LPH-precursor occurs intracellularly, yielding a mature enzyme in the form of a two active site polypeptide which is anchored via a C-terminal peptide. The role of the large cleaved propolypeptide of LPH is not yet known. MGA is the largest of the three disaccharidases, having a molecular weight of greater than 300 kDa. No proteolytic processing seems to be taking place during biogenesis of MGA in human mucosa, and the mode of attachment to the membrane is unknown at present. The application of the methods described to the investigation of congenital sucrase-isomaltase deficiency (CSID) and lactase restriction in adults is presented and differences between CSID and LPH restriction are discussed.     

Molecular aspects of glucocorticoid sensitivity

Glucocorticoids play an essential role in maintaining basal and stress-related homeostasis. Most known effects of glucocorticoids are mediated by the intracellular glucocorticoid receptors. The glucocorticoid sensitivity seems to depend on the amount of receptors expressed and the efficiency of glucocorticoid receptor-mediated signal transduction. Glucocorticoid resistance or hypersensitivity, seen in autoimmune-inflammatory diseases and in metabolic syndrome respectively, can represent the variability of several steps that influence the signaling cascade of glucocorticoid action. The recognition of these steps could provide the understanding of the clinical phenotype and course of such diseases as well as their responsiveness to glucocorticoid therapy. The comprehension of these pathophysiological mechanisms can also improve the possible therapeutic interventions. In this review, we have summarized the multiple factors that have been shown to be involved in this signaling cascade and, thus, to influence glucocorticoid sensitivity.     

Molecular aspects of myeloproliferative neoplasms

During these past 5 years several studies have provided major genetic insights into the pathogenesis of the so-called classical myeloproliferative neoplasms (MPNs): polycythemia vera (PV), essential thrombocythemia (ET), and primary myelofibrosis (PMF). The discovery of the JAK2V617F mutation first, then of the JAK2 exon 12 and MPLW515 mutations, have modified the understanding of these diseases, their diagnosis, and management. Now it is established that almost 100% of PV patients present a JAK2 mutation. Nearly 60% of ET patients and 50% of patients with PMF have the JAK2V617F mutation. The MPLW515 mutations are also present in a small proportion of ET and PMF patients. These mutations are oncogenic events that cause these disorders; however, they do not explain the heterogeneity of the entities in which they occur. Genetic defects have not been yet identified in around 40% of ET and PMF. There are likely additional somatic genetic factors important for the MPN phenotype like the recently described TET2, ASXL1, and CBL mutations. Moreover, polymorphisms in the JAK2 gene have been recently described as associated with MPN. Additional studies of large cohorts are required to dissect the genetic events in MPNs and the mechanisms of these oncogenic cooperations.