Subtypes Sst Research Articles

Inhibition of proliferation of small intestinal and bronchopulmonary neuroendocrine cell lines by using peptide analogs targeting receptors

Currently, no consistently effective therapy is available to inhibit cell proliferation or metastasis of neuroendocrine tumor (NET) disease. The effects of 4 novel peptides were analyzed: a targeted cytotoxic analog of luteinizing hormone-releasing hormone (LH-RH) analog (AN-152), a targeted cytotoxic analog of somatostatin (AN-238), and 2 antagonists of growth hormone-releasing hormone (GH-RH) on 3 NET (carcinoid) cell lines that expressed respective peptide receptors. The effects of the compounds were evaluated on cell proliferation in vitro using MTT uptake and Ki67 expression, apoptosis (caspase 3 expression and activity), and cell cycle parameters (DNA distribution). Proliferation of the LH-RH receptor-expressing lung NET, NCI-H720 line, was inhibited 2-fold by AN-152 containing doxorubicin compared with the chemotherapy alone (IC50 of 9.1 nM vs 24 nM). This was associated with a reduction in Ki67 transcript and an increase in both caspase 3 mRNA levels and activity. Proliferation of the GH-RH receptor expressing lung NET, NCI-H727 line, was inhibited by both GH-RH antagonists, the effects being mediated through changes in Ki67 expression, but not in caspase 3-mediated apoptosis. The small intestinal NET, KRJ-I line, was 8x more sensitive to inhibition by AN-238 than to 2-pyrolino-doxorubicin, reflected by increased caspase 3 transcript as well as activity. AN-238-mediated growth inhibition culminated in complete G1 arrest. The data demonstrate GH-RH antagonists or peptide-linked antineoplastic agents such as AN-152 and AN-238 are effective inhibitors of NET proliferation in vitro. Because peptide receptors such as those for GH-RH, LH-RH, and SST subtypes are commonly expressed by NETs, the development of antineoplastic agents targeted to specific tumor receptors may provide a more efficacious strategy than systemic chemotherapeutic agents currently in use.

Urotensin II and urotensin II-related peptide activate somatostatin receptor subtypes 2 and 5

The UII and urotensin II-related peptide (URP) genes belong to the same superfamily as the somatostatin gene. It has been previously shown that somatostatin activates the UII-receptor (UTR). In contrast, the possible interaction between UII and URP and somatostatin receptors has remained scarcely analyzed. Herein, we have investigated the effects of UII and URP on cell proliferation and free cytosolic Ca2+ concentration ([Ca2+]i) in CHO-K1 cells stably expressing the porcine somatostatin receptor subtypes sst2 and sst5. Results show that both UII and URP induce stimulation of cell proliferation mediated by sst2 receptors and UII provokes inhibition of cell proliferation mediated by sst5 receptors. UII and URP also provoked an increase of [Ca2+]i in both sst2- and sst5-transfected cells. Together, our present data demonstrate that UII and URP directly activate sst2 and sst5 and thus mimic the effect of somatostatin on its cognate receptors.

Quantitative Gene Expression of Somatostatin Receptors and Noradrenaline Transporter Underlying Scintigraphic Results in Patients with Neuroendocrine Tumors

Aim: To measure, by a quantitative approach, the gene expression underlying the results of somatostatin receptor (sst) scintigraphy (¹¹¹In-DTPA-octreotide) and noradrenaline transporter (NAT) scintigraphy (¹²³I-MIBG) in patients with neuroendocrine (NE) tumors. Methods: The gene expression of somatostatin receptors 1–5 (sst) and NAT was measured quantitatively by real-time PCR in a group of patients with NE tumors (n = 14) and compared to a group of patients with colorectal adenocarcinomas (n = 15). If available, scintigraphic results were compared with gene expression results (9 octreotide and 3 MIBG scintigraphies). Results: The sst₂ was upregulated in 13 of 14 patients (93%) with NE tumors, and the absolute level of gene expression was highest for sst₂. Gene expression alterations of NAT and the other sst subtypes were more variable. Gene expression of sst₂ was in all cases in agreement with positive octreotide scintigraphies. In 2 of 3 cases where MIBG scintigraphy was positive, NAT was also upregulated. Sst₂ was generally downregulated in the colorectal tumor group with the gene expression of the other receptors being more heterogeneous. Conclusions: In general, changes in gene expression of sst₂ corresponded with scintigraphic results. Our data support that sst₂ is the best target for visualization of NE tumors, whereas NAT is only a useful target in a subpopulation of NE tumors. Comparison of scintigraphic results with quantitative gene expression may be used to achieve a better understanding of the link between them, which in turn could aid in planning and development of noninvasive molecular imaging of key molecular processes.

Dimerization of G protein-coupled receptors: New avenues for somatostatin receptor signalling, control and functioning

Somatostatin acts through binding and activation of five G protein-coupled receptors (GPCRs) termed somatostatin receptors or ssts (sst1-sst5). These receptors, as many other GPCRs are not just monomers but display a differential tendency to homodimerize, which varies depending on the sst subtype. Moreover, there is evidence that pairs of distinct receptors such as ssst2-sst3 and sst1-sst5 crosstalk by establishing a physical interaction, which results in altered pharmacological or/and functional properties. In addition, ssts can also heterodimerize with other families of GPCRs, as opioid and dopamine receptors, originating heterodimers which properties are different to those of their separated receptors. The present review summarizes the current knowledge on ssts homodimerization, heterodimerization, and interaction with other GPCRs, as well as how interactions affect different aspects of the normal functioning of these receptors.

Somatostatin and somatostatin receptors in Cushing's disease

Cushing's disease is caused by an ACTH secreting pituitary adenoma. Surgery is the treatment of choice and cure rates between 60 and 90% are reported. For patients in which surgery fails, effective medical treatment options are needed. Somatostatin (SS) receptors (sst) are expressed on normal and tumoral corticotroph cells. However, the role of somatostatin and in particular the current clinically available sst(2)-preferring SS analogs in the regulation of normal ACTH secretion, as well as in lowering ACTH and cortisol hypersecretion in patients with Cushing's disease, has been shown to be limited. Recent studies have provided renewed insights into the expression of sst subtypes, as well as into the functional role of SS-analogs in the regulation of ACTH secretion by corticotroph tumors. Sst(2) and sst(5) seem the predominantly expressed sst in corticotroph adenoma cells and targeting both these receptors with a new generation of multiligand SS analogs showed promising effects in terms of lowering ACTH release and urinary free cortisol (UFC) levels in patients with Cushing's disease. In this review an overview of the current insights into the role of SS and sst in the regulation of normal and pathological ACTH secretion is provided.

Intracellular trafficking of somatostatin receptors

The somatostatin receptor subtypes 1-5 (sst(1)-sst(5)) exhibit different intracellular trafficking and endosomal sorting after agonist exposure. The internalization of the somatostatin receptor subtypes sst(2), sst(3) and sst(5) occurs to a much higher extent after somatostatin exposure than of sst(1) or sst(4). After endocytosis, sst(2) and sst(5) recycle to the plasma membrane, whereas sst(3) is predominantly down-regulated. This review will focus on the molecular mechanisms of the differential intracellular trafficking of sst(2), sst(3) and sst(5), and discusses our current knowledge on somatostatin receptor interacting proteins.

Genetic abnormalities of somatostatin receptors in pituitary tumors

Somatostatin exerts antisecretive and antiproliferative effects on different endocrine cells by acting through a family of G protein-coupled receptors that includes five subtypes (SST1-5). Normal human pituitary and pituitary adenomas have been shown to express almost all SST subtypes, with the exception of SST4. Consistent with the observation that octreotide and other somatostatin analogs bind to SST2 and SST5 with high affinity, these genes have been screened for quantitative/qualitative abnormalities in tumors removed from patients with poor responsiveness to somatostatin analogs treatment. Data obtained in GH-secreting adenomas suggested that resistance to octreotide was frequently associated with low expression of SST2 mRNA, although other authors failed to confirm this finding. To date, the only mutational change involving SST2 and SST5 is the Arg to Trp substitution in codon 240 of the SST5 gene that was found in one acromegalic patient resistant to octreotide. Similarly, loss of heterozygosis at SST5 gene locus in pituitary adenomas has been described in individual tumors. In recent years, molecular studies investigated the possible association of gene polymorphisms and susceptibility to diseases and/or resistance to drugs. As far as polymorphic variants of SST genes are concerned, a possible role of SST5 C1004T and T-461C alleles in influencing GH and IGF-I levels in patients with acromegaly has been proposed. Nevertheless, polymorphic variants in SST2 and SST5 genes seem to have a minor, if any, role in determining the different responsiveness to somatostatin analogs in patients with acromegaly.

Alteration of the somatostatinergic system in the striatum of rats with acute experimental autoimmune encephalomyelitis

To date, the neurochemical basis underlying the motor and cognitive deficits described in patients with multiple sclerosis (MS) is unclear. Since the neuropeptide somatostatin (SRIF) and the striatum have been implicated in movement control and implicit memory, the aim of this study was to analyze the striatal somatostatinergic system in an animal model of MS, experimental autoimmune encephalomyelitis (EAE). Female Lewis rats were immunized with an emulsion containing myelin basic protein (MBP) in complete Freund’s adjuvant to induce the disease. The animals were decapitated when limp tail (grade 1) or severe hind limb paralysis (grade 3) was observed. Acute EAE in grade 3 did not modify striatal somatostatin-like immunoreactivity (SRIF-LI) content but decreased the overall SRIF receptor density, without affecting the apparent affinity, in the rat striatal membranes. A selective reduction in the protein levels of the SRIF receptor subtype sst2, analyzed by Western blotting, was detected in the EAE rats, which correlated with decreased sst2 mRNA levels. The expression of the receptor subtypes sst1, sst3 or sst4 was unaltered by the disease. The decrease in the SRIF receptor density was accompanied by an attenuated capacity of SRIF to inhibit both basal and forskolin-stimulated adenylyl cyclase activity. No significant changes, however, were found in the protein levels of Gi proteins (Giα1, Giα2 or Giα3) nor in those of the G-protein-coupled receptor kinase subtypes GRK2, GRK5 or GRK6. Acute EAE in grade 1 did not modify any of the parameters studied. In conclusion, these data demonstrate that acute EAE, in grade 3, disrupts the rat striatal SRIF receptor-effector system. These findings provide new insight into the molecular basis of EAE which might contribute to a better understanding of multiple sclerosis in humans.

Expression and localisation of somatostatin receptor subtypes sst1–sst5 in areas of the rat medulla oblongata involved in autonomic regulation

Somatostatin is known to modulate the activity of neurones of the medulla oblongata involved in autonomic regulation, mediated through five subtypes of G protein-coupled receptors, sst1–sst5. This study utilises reverse transcription polymerase chain reaction and immunohistochemistry to investigate the expression of sst1–sst5, including the sst2 A/sst2 B isoforms, in the main autonomic centres of the rat medulla oblongata: nucleus of the solitary tract (NTS), dorsal motor vagal nucleus (DVN) and ventrolateral medulla (VLM). In tissue from the cerebral cortex, hippocampus and cerebellum all subtype mRNAs were detected, but sst5 signals were weak, and the distribution of sst1–sst5 immunoreactivities was consistent with previous reports. In the medulla, all sst mRNAs gave clear amplicons and subtype-specific antibodies produced characteristic patterns of immunolabelling, frequently in areas of somatostatinergic innervation. Anti-sst1 labelled beaded fibres, sst2 A, sst2 B, sst4 and sst5 gave somatodendritic labelling and sst3 labelled presumptive neuronal cilia. In NTS tissue, sst1, sst2 A, sst4 and sst5 mRNAs were strongly expressed, while in VLM tissue sst1, sst2 A, sst2 B and sst4 predominated. In both areas of the medulla, neurones with intense somatodendritic sst2 A immunoreactivity were principally catecholaminergic in phenotype, being double labelled for tyrosine hydroxylase (TH) and phenylethanolamine- N-methyl-transferase (PNMT). Some TH/PNMT positive neurones were also sst2 B and sst4 immunoreactive. Cholinergic parasympathetic neurones in the DVN were immunoreactive for the sst2 A, sst2 B, sst4 and sst5 subtypes. These observations are consistent with the proposal that multiple somatostatin receptor subtypes, possibly combining as heterodimers, are involved in mediating the modulatory effects of somatostatin on autonomic function, including cardiovascular, respiratory and gastrointestinal reflex activity.

Spontaneous hypersensitivity in mesenteric afferent nerves of mice deficient in the sst2 subtype of somatostatin receptor

Somatostatin is an inhibitory peptide present in abundance in the gastrointestinal (GI) tract. The effects of somatostatin are mediated through its interaction with a family of G-protein-coupled receptors, namely sst1-5. Previous evidence suggested that the sst2 receptor mediates an inhibitory role of somatostatin on GI afferent nerve sensitivity. In the present study we further evaluated mechanical and chemical sensitivity of mesenteric afferents in mice deficient in the sst2 receptor. Multi-unit recordings were made from mesenteric afferents from mouse jejunal segments perfused in vitro. Ramp distension of the jejunum up to 60 mmHg induced biphasic increases in afferent activity in both wild-type (WT) and sst2 gene knock-out (KO) mice. However, the level of afferent activity was significantly higher in the KO (n=15) compared to the WT (n=16) mice across the entire pressure range. The mesenteric afferent sensitivity to acid was evaluated by intraluminal infusion of hydrochloric acid (HCl 20 mM) for 2 min. Peak afferent discharge rate following acid infusion was significantly greater in KO (36.76 +/- 6.47 impulses s(-1), n=7) than in WT preparations (16.53 +/- 3.91 impulses s(-1), n=5, P<0.01). The response to bath-applied bradykinin (1 microm, 3 ml) was not significantly different in the KO and the WT preparations. It is interesting that in the WT preparations, octreotide inhibited both low- and high-threshold mechanosensory responses, whereas in the sst2 KO group it appeared to inhibit the low-threshold responses preferentially and failed to affect the high-threshold responses. The results of the present investigation demonstrate that sst2 deficiency is associated with exaggerated jejunal afferent sensitivity to both mechanical and chemical stimulations, suggesting that somatostatin plays an important inhibitory role in the control of visceral sensitivity by interacting with the sst2 receptor.

Erratum

Correction to: Differential Expression of the Human Somatostatin Receptor Subtypes sst1 to sst5 in Various Adrenal Tumors and Normal Adrenal GlandHorm Metab Res 2005; 37(12): 722-728DOI: 10.1055/s-2005-921092

Somatostatin receptor imaging for neuroendocrine tumors

Tumors and metastases that express the somatostatin receptor subtypes sst2 sst3 or sst5 can be visualized in vivo after injection of radiolabeled octapeptide somatostatin analogs, like (111)In-pentetreotide. (111)In-pentetreotide scintigraphy also allows for more accurate staging of the disease by demonstrating tumor sites, which were not shown by conventional imaging. (111)In-pentetreotide scintigraphy may also detect resectable tumors that would have remained unrecognized using conventional radiological imaging techniques; it may prevent surgery with curative intent in those patients whose tumors have metastasized to a greater extend than could be detected with conventional radiological imaging and it may be used to select patients for treatment with the currently available octapeptide somatostatin analogs or with tumor targeted radioactive treatment with radiolabelled somatostatin analogs. (111)In-pentetreotide scintigraphy has also been used to select patients with pituitary tumors for medical treatment with octapeptide analogs, but its clinical usefulness for this purpose seems to be limited. It further allows scar tissue to be differentiated from tumor recurrence after the pituitary surgery or radiotherapy. However, a large variety of lesions in and around the pituitary region also express somatostatin receptors and, therefore, can be visualized by (111)In-pentetreotide scintigraphy.

Somatostatin Presynaptically Inhibits Both GABA and Glutamate Release Onto Rat Basal Forebrain Cholinergic Neurons

A whole cell patch-clamp study was carried out in slices obtained from young rat brain to elucidate the roles of somatostatin in the modulation of synaptic transmission onto cholinergic neurons in the basal forebrain (BF), a region that contains cholinergic and GABAergic corticopetal neurons and somatostatin (SS)-containing local circuit neurons. Cholinergic neurons within the BF were identified by in vivo prelabeling with Cy3 IgG. Because in many cases SS is contained in GABAergic neurons in the CNS, we investigated whether exogenously applied SS can influence GABAergic transmission onto cholinergic neurons. Bath application of somatostatin (1 muM) reduced the amplitude of the evoked GABAergic inhibitory presynaptic currents (IPSCs) in cholinergic neurons. SS also reduced the frequency of miniature IPSCs (mIPSCs) without affecting their amplitude distribution. SS-induced effect on the mIPSC frequency was significantly larger in the solution containing 7.2 mM Ca(2+) than in the standard (2.4 mM Ca(2+)) external solution. Similar effects were observed in the case of non-NMDA glutamatergic excitatory postsynaptic currents (EPSCs). SS inhibited the amplitude of evoked EPSCs and reduced the frequency of miniature EPSCs dependent on the external Ca(2+) concentration with no effect on their amplitude distribution. Pharmacological analyses using SS-receptor subtype-specific drugs suggest that SS-induced action of the IPSCs is mediated mostly by the sst(2) subtype, whereas sst subtypes mediating SS-induced inhibition of EPSCs are mainly sst(1) or sst(4). These findings suggest that SS presynaptically inhibits both GABA and glutamate release onto BF cholinergic neurons in a Ca(2+)-dependent way, and that SS-induced effect on IPSCs and EPSCs are mediated by different sst subtypes.

Immunohistochemical Distribution and Subcellular Localization of the Somatostatin Receptor Subtype 1 (sst1) in the Rat Hypothalamus

The aim of the present study was to examine the cellular and sub-cellular distribution of the somatostatin (SRIF) receptor subtype sst1 in the rat hypothalamus. Receptors were immunolabeled using an antibody directed against an antigenic sequence in the N-terminus of the receptor. Immunopositive neuronal cell bodies and dendrites were observed throughout the mediobasal hypothalamus, including the medial preoptic area, paraventricular, periventricular, and arcuate nuclei. Immunoreactive axons and axon terminals were also observed in the median eminence, suggesting that sst1 is also located pre-synaptically. Electron microscopic examination of the arcuate nucleus revealed a predominant association of immunoreactive sst1 with perikarya and dendrites. Most immunoreactive receptors were intracellular and localized to tubulovesicular compartments and organelles such as the Golgi apparatus, but 14% were associated with the plasma membrane. Of the latter, 47% were apposed to abbuting afferent axon terminals and 20% localized immediately adjacent to an active synaptic zone. These results demonstrate a widespread distribution of sst1 receptors in rat hypothalamus. They also show that somatodendritic sst1 receptors in the arcuate nucleus are ideally poised to mediate SRIF's modulation of afferent synaptic inputs, including central SRIF effects on growth hormone-releasing hormone neurons documented in this area.

Differential Expression of the Human Somatostatin Receptor Subtypes sst1 to sst5 in Various Adrenal Tumors and Normal Adrenal Gland

Somatostatin (SRIF) is a widely distributed peptide with growth-inhibiting effects in various tumors. So far, five distinct human SRIF receptor subtypes (sst1-sst5) have been identified. We investigated expression of the five ssts in various adrenal tumors and in normal adrenal gland. Tissue was obtained from ten pheochromocytomas (PHEOs), nine cortisol-secreting adenomas (CPAs), eleven aldosterone secreting adenomas (APAs) and eight non-functional adenomas (NFAs) after retroperitoneoscopic surgery, and used for RNA extraction. Adrenal tissue surrounding the tumor was available for analysis in twenty-seven cases. Receptor expression was studied by RT-PCR using sst-specific primers and subsequently confirmed by Southern blotting. Expression of all five receptor subtypes was observed in RNA obtained from normal adrenal gland. Furthermore, each receptor subtype was expressed in more than 50 % of all tumors analyzed. No sst5 expression was found in PHEOs, while sst1 was present in nearly all of these tumors. Only a few of the CPAs expressed subtypes sst1 and sst4. Expression of all five subtypes was distributed equally in APAs. No sst4 was found in any of the NFAs. Differential expression of ssts in various adrenal tumors may point to new aspects in the pathogenesis of these adenomas. Furthermore, the presence of specific ssts could expand the diagnostic and therapeutic strategies during management. New subtype specific analogues of SRIF may be used in the future depending on the type of adrenal tumor and receptor subtype expressed.

Somatostatin Receptor Type 2 Undergoes Plastic Changes in the Human Epileptic Dentate Gyrus

Temporal lobe epilepsy (TLE) is characterized by hippocampal sclerosis together with profound losses and phenotypic changes of different classes of interneurons, including those expressing somatostatin (SRIF). To understand the functional significance of the plasticity of SRIF transmission in TLE, unraveling the status of SRIF receptors is, however, a prerequisite. To address this issue, we characterized expression and distribution of the major SRIF receptor, the sst2 subtype, in hippocampal tissue resected in patients with TLE using complementary neuroanatomic approaches. In patients with hippocampal sclerosis, the number of cells expressing sst2 receptor mRNA as well as sst2 receptor-binding sites and immunoreactivity decreased significantly in the CA1-3, reflecting neuronal loss. By contrast, in the dentate gyrus, sst2 receptor mRNA expression was strongly increased in the granule cell layer, and sst2 receptor-binding sites and immunoreactivity was preserved in the inner but decreased significantly in the outer molecular layer. In this latter region, pronounced changes in SRIF terminal fields were observed. Decreased receptor density in the distal dendrites of granule cells is likely to reflect downregulation of sst2 receptors in response to physiopathologic release of SRIF. Because sst2 receptors have anticonvulsant and antiepileptogenic properties, this phenomenon may contribute to the etiology of TLE seizures.

Somatostatin inhibits insulin secretion from pancreatic B-cells through modulation of R-type Ca2+ channel activity

Background: Somatostatin (SST) inhibits insulin secretion from pancreatic B-cells by modulating the activity of Ca2+ permeable channels. Five pharmacologically distinctive subtypes of voltage-operated calcium channels (VOOCs) are known: L-type [CaV1.1–1.4], P/Q-type [CaV2.1], N-type [CaV2.2; R-type [CaV2.3] and T-type [CaV3.1–3.3]). Recently it has been demonstrated that the R-type VOCC subtype plays an important role at regulating the second phase of insulin secretion. Its involvement in SST-dependent regulation of insulin secretion is not known. Aims: To evaluate whether the R-type channel confers the effects of SST and somatostatin receptor subtype (SSTR)-selective agonists on insulin secretion from pancreatic B-cells. Materials & Methods: The effects of SST were determined in fura2-loaded INS-1 cells, a permanent model of insulin-secreting B-cells, using a fluorescence video imaging system. INS-1 were incubated with SST with or without the specific R-type channel blocker SNX-482 (10 nM) and the levels of intracellular Ca2+ concentration ([Ca2+]i) were determined. The effects of SST and SSTR-selective agonists on insulin secretion, in the presence or absence of SNX-482 were measured by an ELISA. Results: Ca2+ responses in INS-1 cells were clearly detectable at high glucose levels (10 and 20 mM) as compared to 5 mM glucose. SST (100 nM) transiently increased [Ca2+]i within 5min, which declined after 15–30min below the baseline (e.g. 15min: 95±2% of control (set to 100%) (n=8; p<0.05)). Combination of SST with the R-type channel blocker SNX-482 resulted in a smaller transient [Ca2+]i increase followed by a much faster decrease below the baseline. The inhibitory effect of SST and SSTR-selective agonists on 20 mM glucose-induced insulin secretion was strongly reduced in the presence of SNX-482. Conclusion: These data demonstrate that “neuronal“ R-type channel (CaV2.3) plays an important role at mediating the inhibitory effects of somatostatin on insulin secretion.

Discovery of Iodinated Somatostatin Analogues Selective for hsst2 and hsst5 with Excellent Inhibition of Growth Hormone and Prolactin Release from Rat Pituitary Cells

Inhibition of growth hormone (GH) and prolactin (PRL) release from the anterior pituitary gland is mediated through somatostatin receptor subtypes sst2 and sst5. It has been found that somatostatin (SS) analogues that are selective for both receptor subtypes are more effective at inhibiting GH and PRL release than monospecific analogues alone. We synthesized several disulfide-bridged octapeptide SS analogues. Iodinated compounds 7, (4-amino-3-iodo)-d-Phe-c[Cys-Tyr-d-Trp-Lys-Val-Cys]-Thr-NH2, and 9, (4-amino-3-iodo)-d-Phe-c[Cys-(3-iodo)-Tyr-d-Trp-Lys-Val-Cys]-Thr-NH2, were as potent as somatostatin in binding at receptors hsst2 and hsst5 and inhibited GH and PRL release from rat pituitary cells as potently as somatostatin.

Novel Subtype Specific and Universal Somatostatin Analogues: Clinical Potential and Pitfalls

The isolation and purification of somatostatin (SS), exactly 30 years ago, has led to the elucidation of physiologic actions of SS. This cyclic peptide is produced in the hypothalamus, throughout the central nervous system, as well as in most major peripheral organs and inhibits hormone release from the anterior pituitary gland, pancreas and the gastro-intestinal tract. The potent inhibitory actions of SS not only led to the clinical application of this peptide, but also resulted in the development of SS-analogues, among which octreotide and lanreotide are most well known and clinically used for several distinct disorders. Almost ten years ago, five different SS receptor subtypes (sst(1-5)) were identified. These receptor subtypes are variably expressed in distinct tissues and bind with varying affinity to the different SS-analogues, providing an excellent tool to unravel the (patho-) physiological function of the five sst subtypes. Following an overview upon the latest developments in SS receptor physiology and established diagnostic and therapeutic efficacy of SS-analogues in several challenging neuroendocrine disorders, this review predominantly focuses upon the latest developments of (clinically) potential novel sst subtype specific analogues as well as universal binding SS-peptides. Also, the role of potential SS antagonists is assessed. Furthermore, the most recent insights concerning targeted sst-mediated chemo- or radiotherapy are discussed, which offers new therapeutic and diagnostic opportunities for patients harbouring sst-positive neuroendocrine diseases. Finally, acknowledged side effects and possible pitfalls of the use of SS-analogues are discussed.

The multi-ligand somatostatin analogue SOM230 inhibits ACTH secretion by cultured human corticotroph adenomas via somatostatin receptor type 5

Currently, there is no effective medical treatment for patients with pituitary-dependent Cushing's disease. A novel somatostatin (SS) analogue, named SOM230, with high binding affinity to SS receptor subtypes sst(1), sst(2), sst(3) and sst(5) was recently introduced. We compared the in vitro effects of the sst(2)-preferring SS analogue octreotide (OCT) and the multi-ligand SOM230 on ACTH release by human and mouse corticotroph tumour cells. By quantitative RT-PCR the sst subtype expression level was determined in human corticotroph adenomas. In vitro, the inhibitory effect of OCT and SOM230 on ACTH release by dispersed human corticotroph adenoma cells and mouse AtT20 corticotroph adenoma cells was determined. In addition, the influence of dexamethasone on the responsiveness to OCT and SOM230 was studied. Corticotroph adenomas expressed predominantly sst(5) mRNA (six out of six adenomas), whereas sst(2) mRNA expression was detected at significantly lower levels. In a 72 h incubation with 10 nmol/l SOM230, ACTH release was inhibited in three out of five cultures (range -30 to -40%). Ten nmol/l OCT slightly inhibited ACTH release in only one of five cultures (- 28%). In AtT20 cells, expressing sst(2), sst(3) and sst(5), SOM230 inhibited ACTH secretion with high potency (IC(50) 0.2 nmol/l). Dexamethasone (10 nmol/l) pre-treatment did not influence the sensitivity of the cells to the inhibitory effect of SOM230, suggesting that sst(5) is relatively resistant to negative control by glucocorticoids. The selective expression of sst(5) receptors in corticotroph adenomas and the preferential inhibition of ACTH release by human corticotroph adenoma cells by SOM230 in vitro, suggest that SOM230 may have potential in the treatment of patients with pituitary-dependent Cushing's disease.