RESUMEN

Sphingosine 1-phosphate (S1P) mediates critical physiological responses by its binding to G protein-coupled receptor (GPCR) subtypes, known as S1P receptors. Five distinct mammalian S1P receptors, designated S1P1-5 have been identified, each with a different cellular pattern of expression which influences the responses to S1P. In this review, we briefly outline our understanding of the modes of action and the roles of S1P receptors in the regulation of physiological and pathological functions in the cardiovascular, immune and central nervous system.

Asunto(s)

Lisofosfolípidos/fisiología , Receptores de Lisoesfingolípidos/fisiología , Esfingosina/análogos & derivados , Animales , Aterosclerosis/fisiopatología , Sistema Cardiovascular/fisiopatología , Movimiento Celular , Humanos , Linfocitos/metabolismo , Activación de Macrófagos , Macrófagos/inmunología , Macrófagos/metabolismo , Macrófagos/ultraestructura , Mamíferos , Ratones , Ratones Noqueados , Neoplasias/irrigación sanguínea , Neoplasias/inmunología , Neovascularización Patológica/fisiopatología , Neovascularización Fisiológica/fisiología , Proteínas del Tejido Nervioso/fisiología , Neurogénesis/fisiología , Receptores de Lisoesfingolípidos/clasificación , Receptores de Lisoesfingolípidos/deficiencia , Receptores de Lisoesfingolípidos/efectos de los fármacos , Sistemas de Mensajero Secundario/fisiología , Esfingosina/fisiología

RESUMEN

Sphingosine-1-phosphate (S1P) is a lysophospholipid signaling molecule that regulates important biological functions in both intracellular and extracellular compartments. It interacts with five G protein-coupled receptors subtypes (S1PR(1-5)) to generate multiple downstream signaling. Activation of S1PR1 has been validated to be involved in the process of immune modulation. Fingolimod (FTY720), the novel S1PR1 agonist, has been approved for the treatment of multiple sclerosis in clinical trials. The study towards discovery of selective S1PR1 agonists has become hot spot for immunological diseases. This article summarized the research progress of S1PR1 agonists, emphasizing their structure types, structure-activity relationship and direction of development.

Asunto(s)

Inmunosupresores/uso terapéutico , Esclerosis Múltiple/tratamiento farmacológico , Glicoles de Propileno/uso terapéutico , Receptores de Lisoesfingolípidos/agonistas , Receptores de Lisoesfingolípidos/fisiología , Esfingosina/análogos & derivados , Animales , Clorhidrato de Fingolimod , Humanos , Inmunosupresores/farmacología , Lisofosfolípidos/fisiología , Glicoles de Propileno/farmacología , Receptores de Lisoesfingolípidos/clasificación , Receptores de Lisoesfingolípidos/metabolismo , Esfingosina/farmacología , Esfingosina/fisiología , Esfingosina/uso terapéutico , Relación Estructura-Actividad

RESUMEN

Sphingosine-1-phospate (S1P) and S1P receptor agonists elicit mechanism-based effects on cardiovascular function in vivo. Indeed, FTY720 (non-selective S1P(X) receptor agonist) produces modest hypertension in patients (2-3 mmHg in 1-yr trial) as well as acute bradycardia independent of changes in blood pressure. However, the precise receptor subtypes responsible is controversial, likely dependent upon the cardiovascular response in question (e.g. bradycardia, hypertension), and perhaps even species-dependent since functional differences in rodent, rabbit, and human have been suggested. Thus, we characterized the S1P receptor subtype specificity for each compound in vitro and, in vivo, the cardiovascular effects of FTY720 and the more selective S1P1,5 agonist, BAF312, were tested during acute i.v. infusion in anesthetized rats and after oral administration for 10 days in telemetry-instrumented conscious rats. Acute i.v. infusion of FTY720 (0.1, 0.3, 1.0 mg/kg/20 min) or BAF312 (0.5, 1.5, 5.0 mg/kg/20 min) elicited acute bradycardia in anesthetized rats demonstrating an S1P1 mediated mechanism-of-action. However, while FTY720 (0.5, 1.5, 5.0 mg/kg/d) elicited dose-dependent hypertension after multiple days of oral administration in rat at clinically relevant plasma concentrations (24-hr mean blood pressure = 8.4, 12.8, 16.2 mmHg above baseline vs. 3 mmHg in vehicle controls), BAF312 (0.3, 3.0, 30.0 mg/kg/d) had no significant effect on blood pressure at any dose tested suggesting that hypertension produced by FTY720 is mediated S1P3 receptors. In summary, in vitro selectivity results in combination with studies performed in anesthetized and conscious rats administered two clinically tested S1P agonists, FTY720 or BAF312, suggest that S1P1 receptors mediate bradycardia while hypertension is mediated by S1P3 receptor activation.

Asunto(s)

Azetidinas/efectos adversos , Compuestos de Bencilo/efectos adversos , Bradicardia/inducido químicamente , Hipertensión/inducido químicamente , Glicoles de Propileno/efectos adversos , Receptores de Lisoesfingolípidos/agonistas , Esfingosina/análogos & derivados , Animales , Azetidinas/farmacología , Compuestos de Bencilo/farmacología , Bradicardia/patología , Células Cultivadas , Evaluación Preclínica de Medicamentos , Clorhidrato de Fingolimod , Humanos , Hipertensión/patología , Inmunosupresores/efectos adversos , Inmunosupresores/farmacología , Masculino , Glicoles de Propileno/farmacología , Ratas , Ratas Sprague-Dawley , Receptores de Lisoesfingolípidos/clasificación , Esfingosina/efectos adversos , Esfingosina/farmacología , Especificidad por Sustrato

RESUMEN

Sphingosine-1-phosphate (S1P) and lysophosphatidic acid (LPA) are endogenous bioactive lipids which mediate a variety of biological cell responses such as cell proliferation, migration, differentiation and apoptosis. Their actions are mediated by binding to the G-protein-coupled endothelial differentiation gene (Edg) receptor subfamily, referred to as S1P1-5 and LPA1-5, and regulate a variety of signalling pathways involved in numerous physiological processes and pathological conditions. Their importance during embryogenesis has been demonstrated by the generation of knock-out mice and specific roles have been assigned to these receptors. However, potential functional redundancy and the lethality of some mutants have complicated functional analysis in these models. Here we report the cloning of the S1P and LPA receptors in Xenopus laevis and tropicalis. Phylogenetic analyses demonstrate the high level of conservation of these receptors between amphibian and other vertebrate species. We have conducted a comparative expression analysis of these receptors during development and in the adult frog, by both RT-PCR and whole mount in situ hybridisation. In particular, we show that S1P1, 2 and 5 display distinct embryonic specific expression patterns, suggesting potentially different developmental roles for these receptors, and therefore for their ligands, during amphibian embryogenesis.

Asunto(s)

Familia de Multigenes , Receptores del Ácido Lisofosfatídico/genética , Receptores de Lisoesfingolípidos/genética , Proteínas de Xenopus/genética , Xenopus laevis/genética , Secuencia de Aminoácidos , Animales , Clonación Molecular , Embrión no Mamífero/embriología , Embrión no Mamífero/metabolismo , Femenino , Perfilación de la Expresión Génica , Regulación del Desarrollo de la Expresión Génica , Hibridación in Situ , Masculino , Datos de Secuencia Molecular , Oocitos/metabolismo , Filogenia , Isoformas de Proteínas/clasificación , Isoformas de Proteínas/genética , Receptores del Ácido Lisofosfatídico/clasificación , Receptores de Lisoesfingolípidos/clasificación , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Homología de Secuencia de Aminoácido , Especificidad de la Especie , Xenopus/embriología , Xenopus/genética , Xenopus laevis/embriología

RESUMEN

Evidence is emerging indicating that sphingosine-1-phosphate (S1P) participates in signaling in the retina. To determine whether S1P might be involved in signaling in the inner retina specifically, we examine the effects of this sphingolipid on cultured retinal amacrine cells. Whole cell voltage-clamp recordings reveal that S1P activates a cation current that is dependent on signaling through G(i) and phospholipase C. These observations are consistent with the involvement of members of the S1P receptor family of G-protein-coupled receptors in the production of the current. Immunocytochemistry and PCR amplification provide evidence for the expression of S1P1R and S1P3R in amacrine cells. The receptor-mediated channel activity is shown to be highly sensitive to blockade by lanthanides consistent with the behavior of transient receptor potential canonical (TRPC) channels. PCR products amplified from amacrine cells reveal that TRPCs 1 and 3-7 channel subunits have the potential to be expressed. Because TRPC channels provide a Ca(2+) entry pathway, we asked whether S1P caused cytosolic Ca(2+) elevations in amacrine cells. We show that S1P-dependent Ca(2+) elevations do occur in these cells and that they might be mediated by S1P1R and S1P3R. The Ca(2+) elevations are partially due to release from internal stores, but the largest contribution is from influx across the plasma membrane. The effect of inhibition of sphingosine kinase suggests that the production of cytosolic S1P underlies the sustained nature of the Ca(2+) elevations. Elucidation of the downstream effects of these signals will provide clues to the role of S1P in regulating inner retinal function.

Asunto(s)

Células Amacrinas/fisiología , Señalización del Calcio/fisiología , Receptores de Lisoesfingolípidos/metabolismo , Células Amacrinas/efectos de los fármacos , Animales , Calcio/metabolismo , Señalización del Calcio/efectos de los fármacos , Células Cultivadas , Embrión de Pollo , Estrenos/farmacología , Heparina/farmacología , Receptores de Inositol 1,4,5-Trifosfato/antagonistas & inhibidores , Activación del Canal Iónico/efectos de los fármacos , Activación del Canal Iónico/fisiología , Elementos de la Serie de los Lantanoides/farmacología , Lisofosfolípidos/farmacología , Potenciales de la Membrana/efectos de los fármacos , Potenciales de la Membrana/fisiología , Modelos Biológicos , Oxadiazoles/farmacología , Técnicas de Placa-Clamp/métodos , Toxina del Pertussis/farmacología , Inhibidores de Fosfodiesterasa/farmacología , Bloqueadores de los Canales de Potasio/farmacología , Pirrolidinonas/farmacología , Ratas , Ratas Sprague-Dawley , Receptores de Lisoesfingolípidos/clasificación , Retina/citología , Esfingosina/análogos & derivados , Esfingosina/farmacología , Tetraetilamonio/farmacología , Tiofenos/farmacología , Fosfolipasas de Tipo C/metabolismo

RESUMEN

OBJECTIVE: The role of sphingosine-1-phosphate (S1P) receptors in acute vascular injury and smooth muscle cell (SMC) phenotypic modulation is not completely resolved. METHODS AND RESULTS: S1P receptor antagonists were used to test the hypothesis that specific S1P receptor subtypes differentially regulate SMC phenotypic modulation. In response to acute balloon injury of the rat carotid artery, S1P1/S1P3 receptor mRNA levels were transiently increased at 48 hours whereas S1P2 receptor expression was decreased. S1P2 expression was reinduced and increased at 7 to 10 days postinjury. Daily intraperitoneal injection of the S1P1/S1P3 antagonist VPC44116 decreased neointimal hyperplasia by approximately 50%. In vitro, pharmacological inhibition of S1P1/S1P3 receptors with VPC25239 attenuated S1P-induced proliferation of rat aortic SMCs. Conversely, inhibition of S1P2 with JTE013 potentiated S1P-induced proliferation. Inhibition of S1P1/S1P3 resulted in S1P-induced activation of the SMC differentiation marker genes SMalpha-actin and SMMHC, whereas inhibition of S1P2 attenuated this response. S1P2-dependent activation of SMalpha-actin and SMMHC was shown to be mediated by L-type voltage-gated Ca(2+) channels and subsequent RhoA/Rho kinase-dependent SRF enrichment of CArG box promoter regions. CONCLUSIONS: Results provide evidence that S1P1/S1P3 receptors promote, whereas S1P2 receptors antagonize, SMC proliferation and phenotypic modulation in vitro in response to S1P, or in vivo after vascular injury.

Asunto(s)

Lisofosfolípidos/fisiología , Miocitos del Músculo Liso/fisiología , Receptores de Lisoesfingolípidos/fisiología , Esfingosina/análogos & derivados , Animales , Masculino , Miocitos del Músculo Liso/clasificación , Fenotipo , Ratas , Receptores de Lisoesfingolípidos/clasificación , Esfingosina/fisiología

RESUMEN

Phylogenetic analysis of transmembrane regions of GPCRs using PHYLIP indicated that the orphan receptor P2Y(10) receptor was classified into the cluster consisting nucleotide and lipid receptors. Based on the results, we studied the abilities of nucleotides and lipids to activate the P2Y(10) receptors. As a result, sphingosine-1-phosphate (S1P) and lysophosphatidic acid (LPA) evoked intracellular Ca(2+) increases in the CHO cells stably expressing the P2Y(10) fused with a G(16alpha) protein. These Ca(2+) responses were inhibited by S1P receptor and LPA receptor antagonists. The introduction of siRNA designed for P2Y(10) receptor into the P2Y(10)-CHO cells effectively blocked both S1P- and LPA-induced Ca(2+) increases. RT-PCR analysis showed that the mouse P2Y(10) was expressed in reproductive organs, brain, lung and skeletal muscle, suggesting the receptor plays physiological roles throughout the whole body. In conclusion, the P2Y(10) receptor is the first receptor identified as a dual lysophospholipid receptor.

Asunto(s)

Receptores del Ácido Lisofosfatídico/clasificación , Receptores del Ácido Lisofosfatídico/fisiología , Receptores de Lisoesfingolípidos/clasificación , Receptores de Lisoesfingolípidos/fisiología , Receptores Purinérgicos P2/clasificación , Receptores Purinérgicos P2/fisiología , Secuencia de Aminoácidos , Animales , Células CHO , Calcio/metabolismo , Cricetinae , Cricetulus , Humanos , Ligandos , Lisofosfolípidos/metabolismo , Lisofosfolípidos/farmacología , Ratones , Datos de Secuencia Molecular , Filogenia , Conformación Proteica , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/farmacología , Receptores del Ácido Lisofosfatídico/genética , Receptores de Lisoesfingolípidos/genética , Receptores Purinérgicos P2/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Análisis de Secuencia de Proteína , Esfingosina/análogos & derivados , Esfingosina/metabolismo , Esfingosina/farmacología , Distribución Tisular