Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 848
Filtrar
1.
Nat Commun ; 10(1): 3196, 2019 07 19.
Artigo em Inglês | MEDLINE | ID: mdl-31324767

RESUMO

The limited clinical response observed in high-grade serous ovarian cancer (HG-SOC) with high frequency of TP53 mutations (mutp53) might be related to mutp53-driven oncogenic pathway network. Here we show that ß-arrestin1 (ß-arr1), interacts with YAP, triggering its cytoplasmic-nuclear shuttling. This interaction allows ß-arr1 to recruit mutp53 to the YAP-TEAD transcriptional complex upon activation of endothelin-1 receptors (ET-1R) in patient-derived HG-SOC cells and in cell lines bearing mutp53. In parallel, ß-arr1 mediates the ET-1R-induced Trio/RhoA-dependent YAP nuclear accumulation. In the nucleus, ET-1 through ß-arr1 orchestrates the tethering of YAP and mutp53 to YAP/mutp53 target gene promoters, including EDN1 that ensures persistent signals. Treatment of patient-derived xenografts reveals synergistic antitumoral and antimetastatic effects of the dual ET-1R antagonist macitentan in combination with cisplatinum, shutting-down the ß-arr1-mediated YAP/mutp53 transcriptional programme. Furthermore, ETAR/ß-arr1/YAP gene signature correlates with a worst prognosis in HG-SOC. These findings support effective combinatorial treatment for repurposing the ET-1R antagonists in HG-SOC.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Cistadenocarcinoma Seroso/metabolismo , Neoplasias Ovarianas/metabolismo , Receptor de Endotelina A/metabolismo , Transdução de Sinais , Fatores de Transcrição/metabolismo , Proteína Supressora de Tumor p53/metabolismo , beta-Arrestina 1/metabolismo , Animais , Antineoplásicos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Cistadenocarcinoma Seroso/tratamento farmacológico , Cistadenocarcinoma Seroso/genética , Modelos Animais de Doenças , Endotelina-1/metabolismo , Feminino , Regulação Neoplásica da Expressão Gênica , Fatores de Troca do Nucleotídeo Guanina/metabolismo , Humanos , Camundongos Nus , Mutação , Neoplasias Ovarianas/tratamento farmacológico , Neoplasias Ovarianas/genética , Proteínas Serina-Treonina Quinases/metabolismo , Pirimidinas/farmacologia , Receptor de Endotelina A/efeitos dos fármacos , Sulfonamidas/farmacologia , Proteína Supressora de Tumor p53/genética , Ensaios Antitumorais Modelo de Xenoenxerto , beta-Arrestina 1/efeitos dos fármacos , Proteínas rho de Ligação ao GTP/metabolismo , Proteína rhoA de Ligação ao GTP/metabolismo
2.
Endocrinology ; 160(8): 1786-1796, 2019 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-31173072

RESUMO

Bone metastasis is a painful complication of advanced prostate cancer. Endothelin-1 is a tumor-secreted factor that plays a central role in osteoblast activation and the osteosclerotic response of prostate cancer metastatic to bone. Antagonists that block the activation of the endothelin A receptor (ETAR), located on osteoblasts, reduce osteoblastic bone lesions in animal models of bone metastasis. However, ETAR antagonists demonstrated limited efficacy in clinical trials of men with advanced prostate cancer who also received standard androgen deprivation therapy (ADT). Previous data from our group suggested that, in a mouse model, ETAR antagonists might only be efficacious when androgen signaling in the osteoblast is lowered beyond the ability of standard ADT. This notion was tested in a mouse model of prostate cancer bone metastasis. Castrated and sham-operated male athymic nude mice underwent intracardiac inoculation of the ARCaPM castration-resistant prostate cancer cell line. The mice were then treated with either the ETAR antagonist zibotentan or a vehicle control to generate four experimental groups: vehicle+sham (Veh+Sham), vehicle+castrate (Veh+Castr), zibotentan+sham (Zibo+Sham), and zibotentan+castrate (Zibo+Castr). The mice were monitored radiographically for the development of skeletal lesions. The Zibo+Castr group had significantly longer survival and a single incidental lesion. Mice in the Zibo+Sham group had the shortest survival and the largest number of skeletal lesions. Survival and skeletal lesions of the Veh+Sham and Veh+Castr groups were intermediate compared with the zibotentan-treated groups. We report a complex interaction between ETAR and androgen signaling, whereby ETAR blockade was most efficacious when combined with complete androgen deprivation.


Assuntos
Neoplasias Ósseas/secundário , Antagonistas do Receptor de Endotelina A/uso terapêutico , Neoplasias da Próstata/tratamento farmacológico , Pirrolidinas/uso terapêutico , Animais , Linhagem Celular Tumoral , Modelos Animais de Doenças , Endotelina-1/sangue , Masculino , Camundongos , Orquiectomia , Osteoblastos/efeitos dos fármacos , Osteoblastos/fisiologia , Neoplasias da Próstata/mortalidade , Neoplasias da Próstata/patologia , Receptor de Endotelina A/metabolismo
3.
Oxid Med Cell Longev ; 2019: 6508328, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31214281

RESUMO

Endothelin-1 (ET-1) is synthesized primarily by endothelial cells. ET-1 administration in vivo enhances the cardiac sympathetic afferent reflex and sympathetic activity. Previous studies have shown that sympathetic hyperactivity promotes malignant ventricular arrhythmia (VA). The aim of this study was to investigate whether ET-1 could activate the left stellate ganglion (LSG) and promote malignant VA. Twelve male beagle dogs who received local microinjections of saline (control, n = 6) and ET-1 into the LSG (n = 6) were included. The ventricular effective refractory period (ERP), LSG function, and LSG activity were measured at different time points. VA was continuously recorded for 1 h after left anterior descending occlusion (LADO), and LSG tissues were then collected for molecular detection. Compared to that of the control group, local ET-1 microinjection significantly decreased the ERP and increased the occurrence of VA. In addition, local microinjection of ET-1 increased the function and activity of the LSG in the normal and ischemic hearts. The expression levels of proinflammatory cytokines and the protein expression of c-fos and nerve growth factor (NGF) in the LSG were also increased. More importantly, endothelin A receptor (ETA-R) expression was found in the LSG, and its signaling was significantly activated in the ET-1 group. LSG activation induced by local ET-1 microinjection aggravates LADO-induced VA. Activated ETA-R signaling and the upregulation of proinflammatory cytokines in the LSG may be responsible for these effects.


Assuntos
Arritmias Cardíacas/fisiopatologia , Células Endoteliais/metabolismo , Endotelina-1/metabolismo , Isquemia Miocárdica/fisiopatologia , Gânglio Estrelado/metabolismo , Animais , Células Cultivadas , Citocinas/metabolismo , Modelos Animais de Doenças , Cães , Eletrocardiografia , Células Endoteliais/patologia , Humanos , Mediadores da Inflamação/metabolismo , Masculino , Fatores de Crescimento Neural/metabolismo , Receptor de Endotelina A/metabolismo , Transdução de Sinais , Sistema Nervoso Simpático
4.
Life Sci ; 228: 295-304, 2019 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-31075232

RESUMO

AIMS: To investigate the protective effects of downregulating ETaR expression on renal ischemia reperfusion injury (IRI). MAIN METHODS: The renal IRI model was generated by clamping the left renal artery for 60 min followed by nephrectomy of the right kidney. ETaR siRNA were perfused through the renal artery during ischemia. HE staining was performed to assess histological injury. PCR was performed to determine the expression of NF-κb, TNF-α, IFN-γ, IL-6 and TGF-ß. ELISA was used to determine the levels of ET-1, TGF-ß and eNOS. The level of nitric oxide (NO) was tested by the NO detection kit. The expression of PI3K, Akt, sGC and PKG were evaluated by western blot. KEY FINDINGS: ETaR siRNA treatment reduced the levels of serum creatinine and urea nitrogen, decreased the number of apoptotic cells, and ameliorated histological damage after IRI. PCR results demonstrated that IRI increased mRNA levels of inflammatory factors, which were inhibited by ETaR siRNA treatment. ELISA result showed that ETaR siRNA decreased the levels of ET-1, TGF-ß and eNOS in the renal tissues after IRI. Western blot results demonstrated that ETaR siRNA activated the PI3K/Akt and sGC/PKG signaling pathway. Conversely, the NOS inhibitor, L-NAME, reversed the effects of ETaR siRNA treatment. SIGNIFICANCE: ETaR siRNA treatment inhibited inflammatory response and improved renal function after renal IRI. The underlying mechanisms of ETaR siRNA treatment may be through increasing eNOS activity through PI3K/Akt signaling, which subsequently increased NO production. The increased NO reduces the expression of ET-1 by inhibiting transcription of ET-1-associated genes via the sGC/PKG signaling pathway.


Assuntos
Rim/patologia , Óxido Nítrico/metabolismo , RNA Interferente Pequeno/uso terapêutico , Terapêutica com RNAi , Receptor de Endotelina A/genética , Traumatismo por Reperfusão/genética , Traumatismo por Reperfusão/terapia , Animais , Apoptose , Rim/metabolismo , Masculino , Óxido Nítrico Sintase Tipo III/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , RNA Interferente Pequeno/genética , Terapêutica com RNAi/métodos , Ratos Sprague-Dawley , Receptor de Endotelina A/metabolismo , Traumatismo por Reperfusão/metabolismo , Traumatismo por Reperfusão/patologia , Transdução de Sinais
5.
J Pharmacol Sci ; 140(1): 102-105, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31103330

RESUMO

Endothelin type A receptor (ETAR) is internalized upon agonist stimulation; however, the mechanism thereof remains controversial. In this study, we characterized the endothelin-1 (ET-1)-induced internalization of ETAR expressed in Chinese hamster ovary cells. ET-1 elicited ETAR internalization and increase in intracellular Ca2+ concentration. ET-1-induced ETAR internalization was completely inhibited by a reduction in intracellular and extracellular Ca2+ levels and partially suppressed by inhibitors of protein kinase C (PKC) and extracellular signal-regulated kinases 1/2 (ERK1/2), both of which are downstream molecules in ETAR signaling. These results suggest that Ca2+ mobilization, PKC, and ERK1/2 are involved in ET-1-induced ETAR internalization.


Assuntos
Sinalização do Cálcio/fisiologia , Endotelina-1/farmacologia , Receptor de Endotelina A/metabolismo , Animais , Células CHO , Cálcio/metabolismo , Cricetinae , Cricetulus , Feminino , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/fisiologia , Proteína Quinase C/metabolismo , Proteína Quinase C/fisiologia , Transdução de Sinais/efeitos dos fármacos
6.
J Int Med Res ; 47(5): 2177-2186, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30975046

RESUMO

OBJECTIVES: To assess whether switching patients with suboptimally controlled pulmonary arterial hypertension from bosentan or ambrisentan to macitentan would improve six-minute walk test (6MWT) distance and World Health Organization functional class. METHODS: This was a retrospective cohort analysis of 37 patients from a single center. Patients were separated into three heterogeneous treatment groups and followed for 18 months: switch group (n = 14): patients switched to macitentan from bosentan/ambrisentan; added group (n = 11): patients who began macitentan as de novo therapy (n = 5) or who added macitentan to an existing sildenafil regimen (n = 6); and control group (n = 12): patients for whom sildenafil and/or bosentan/ambrisentan therapy was unchanged. RESULTS: Mortality was observed in two patients (one each, switch and added groups). Patients in the control group had one hospital admission and 100% survival. There was significant improvement in functional class for the switch and added groups. Statistically significant improvement was observed in 6MWT distance in the added group alone. Overall, 92% of patients continued macitentan throughout the study. CONCLUSION: Macitentan was well tolerated. For bosentan/ambrisentan-treated patients with suboptimally controlled pulmonary arterial hypertension, switching to macitentan may facilitate an improvement in functional class.


Assuntos
Antagonistas do Receptor de Endotelina A/uso terapêutico , Hipertensão Pulmonar/tratamento farmacológico , Pirimidinas/uso terapêutico , Receptor de Endotelina A/metabolismo , Sulfonamidas/uso terapêutico , Idoso , Antagonistas do Receptor de Endotelina A/efeitos adversos , Feminino , Humanos , Hipertensão Pulmonar/diagnóstico por imagem , Hipertensão Pulmonar/fisiopatologia , Masculino , Pessoa de Meia-Idade , Pirimidinas/efeitos adversos , Sulfonamidas/efeitos adversos , Sístole/efeitos dos fármacos , Resultado do Tratamento , Função Ventricular/efeitos dos fármacos , Teste de Caminhada , Organização Mundial da Saúde
7.
Biochim Biophys Acta Gen Subj ; 1863(5): 917-924, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30851407

RESUMO

The angiotensin II AT1 and the endothelin 1 ETA receptors play a crucial role in the pathogenesis of cardiovascular diseases like hypertension, heart failure, stroke, pulmonary hypertension, and cardiac hypertrophy. Both receptors are members of the rhodopsion-like superfamily of G protein-coupled receptors which can exist as monomers, dimers, and higher order aggregates. Recently, oligomerization of these two receptors have been described by several biophysical methods based mainly on luminescence and fluorescence energy transfer. Since this oligomerization can occur either spontaneously or it can be induced by ligand-binding, the aim of this work was to address whether the oligomerization of these receptors occurs upon ligand-binding. For this purpose the Number and Brightness analysis, a method that allows the identification, localization, and quantification of protein aggregates in the plasma membrane of a single cell, was used. An advantage of this method is that it is not limited to certain dyes specially required for Fluorescence Resonance Energy Transfer measurements. Our results showed that stably transfected angiotensin II AT1 receptors and transiently transfected endothelin 1 ETA receptors, were found as monomeric, dimeric, and tetrameric receptor aggregates. Interestingly, the binding of antihypertensive agents like losartan and BQ123, earlier suggested to be inverse agonists, significantly increased the proportion of monomers and reduced the occurrence of dimers on the cell membrane; while the kown endothelin 1 ETA antagonist sitaxentan did not influence the aggregation state of these receptors.


Assuntos
Angiotensina II/metabolismo , Endotelina-1/agonistas , Receptor de Endotelina A/metabolismo , Animais , Sítios de Ligação/efeitos dos fármacos , Células CHO , Cricetulus , Endotelina-1/metabolismo , Transferência Ressonante de Energia de Fluorescência , Humanos , Isoxazóis/farmacologia , Ligantes , Losartan/farmacologia , Peptídeos Cíclicos/farmacologia , Agregados Proteicos/efeitos dos fármacos , Tiofenos/farmacologia
8.
Nat Commun ; 10(1): 1387, 2019 03 27.
Artigo em Inglês | MEDLINE | ID: mdl-30918259

RESUMO

Inhibitors of the secretion of cancer exosomes, which promote cancer progression and metastasis, may not only accelerate exosome biology research but also offer therapeutic benefits for cancer patients. Here we identify sulfisoxazole (SFX) as an inhibitor of small extracellular vesicles (sEV) secretion from breast cancer cells through interference with endothelin receptor A (ETA). SFX, an FDA-approved oral antibiotic, showed significant anti-tumor and anti-metastatic effects in mouse models of breast cancer xenografts, the reduced expression of proteins involved in biogenesis and secretion of sEV, and triggered co-localization of multivesicular endosomes with lysosomes for degradation. We demonstrate the important role of ETA, as target of SFX, by gain- and loss-of-function studies of the ETA protein, through a direct binding assay, and pharmacological and genetic approaches. These findings may provide a foundation for sEV-targeted cancer therapies and the mechanistic studies on sEV biology.


Assuntos
Anti-Infecciosos/farmacologia , Neoplasias da Mama/metabolismo , Vesículas Extracelulares/efeitos dos fármacos , Receptor de Endotelina A/efeitos dos fármacos , Sulfisoxazol/farmacologia , Animais , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Endossomos/efeitos dos fármacos , Endossomos/metabolismo , Vesículas Extracelulares/metabolismo , Feminino , Humanos , Lisossomos/efeitos dos fármacos , Lisossomos/metabolismo , Células MCF-7 , Masculino , Camundongos , Camundongos Nus , Metástase Neoplásica , Biogênese de Organelas , Receptor de Endotelina A/metabolismo , Ensaios Antitumorais Modelo de Xenoenxerto
9.
Am J Physiol Renal Physiol ; 316(6): F1173-F1179, 2019 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-30864842

RESUMO

Emerging evidence indicates that endogenous production of endothelin (ET)-1, a 21-amino acid peptide vasoconstrictor, plays an important role in proteinuric kidney disease. Previous studies in rats have shown that chronic administration of ET-1 leads to increased glomerular albumin leakage. The underlying mechanisms are, however, currently not known. Here, we used size-exclusion chromatography to measure glomerular sieving coefficients for neutral FITC-Ficoll (molecular Stokes-Einstein radius: 15-80 Å, molecular weight: 70 kDa/400 kDa) in anesthetized male Sprague-Dawley rats (n = 12) at baseline and at 5, 15, 30, and 60 min after intravenous administration of ET-1. In separate experiments, ET-1 was given together with the selective ET type A (ETA) or ET type B (ETB) receptor antagonists JKC-301 and BQ-788, respectively. At both 15 and 30 min postadministration, the glomerular sieving coefficient for macromolecular Ficoll (70 Å) was significantly increased to 4.4 × 10-5 ± 0.7 × 10-5 (P = 0.024) and 4.5 × 10-5 ± 0.8 × 10-5 (P = 0.007), respectively, compared with baseline (2.2 × 10-5 ± 0.4 ×10-5). Decreased urine production after ET-1 prevented the use of higher doses of ET-1. Data analysis using the two-pore model indicated changes in large-pore permeability after ET-1, with no changes in the small-pore pathway. Administration of ETA blocker abrogated the permeability changes induced by ET-1 at 30 min, whereas blockade of ETB receptors was ineffective. Mean arterial pressure was only significantly increased at 60 min, being 123 ± 4 mmHg compared with 111 ± 2 mmHg at baseline (P = 0.02). We conclude that ET-1 evoked small, delayed, and sustained increases in glomerular permeability, mediated via the ETA receptor.


Assuntos
Endotelina-1/administração & dosagem , Taxa de Filtração Glomerular/efeitos dos fármacos , Glomérulos Renais/efeitos dos fármacos , Receptor de Endotelina A/agonistas , Animais , Hemodinâmica/efeitos dos fármacos , Infusões Intravenosas , Glomérulos Renais/metabolismo , Masculino , Modelos Biológicos , Permeabilidade , Ratos Sprague-Dawley , Receptor de Endotelina A/metabolismo , Transdução de Sinais , Fatores de Tempo
10.
PLoS Genet ; 15(2): e1007941, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30811380

RESUMO

Skin pigment patterns are important, being under strong selection for multiple roles including camouflage and UV protection. Pigment cells underlying these patterns form from adult pigment stem cells (APSCs). In zebrafish, APSCs derive from embryonic neural crest cells, but sit dormant until activated to produce pigment cells during metamorphosis. The APSCs are set-aside in an ErbB signaling dependent manner, but the mechanism maintaining quiescence until metamorphosis remains unknown. Mutants for a pigment pattern gene, parade, exhibit ectopic pigment cells localised to the ventral trunk, but also supernumerary cells restricted to the Ventral Stripe. Contrary to expectations, these melanocytes and iridophores are discrete cells, but closely apposed. We show that parade encodes Endothelin receptor Aa, expressed in the blood vessels, most prominently in the medial blood vessels, consistent with the ventral trunk phenotype. We provide evidence that neuronal fates are not affected in parade mutants, arguing against transdifferentiation of sympathetic neurons to pigment cells. We show that inhibition of BMP signaling prevents specification of sympathetic neurons, indicating conservation of this molecular mechanism with chick and mouse. However, inhibition of sympathetic neuron differentiation does not enhance the parade phenotype. Instead, we pinpoint ventral trunk-restricted proliferation of neural crest cells as an early feature of the parade phenotype. Importantly, using a chemical genetic screen for rescue of the ectopic pigment cell phenotype of parade mutants (whilst leaving the embryonic pattern untouched), we identify ErbB inhibitors as a key hit. The time-window of sensitivity to these inhibitors mirrors precisely the window defined previously as crucial for the setting aside of APSCs in the embryo, strongly implicating adult pigment stem cells as the source of the ectopic pigment cells. We propose that a novel population of APSCs exists in association with medial blood vessels, and that their quiescence is dependent upon Endothelin-dependent factors expressed by the blood vessels.


Assuntos
Células-Tronco Adultas/citologia , Células-Tronco Adultas/metabolismo , Receptores ErbB/metabolismo , Pigmentos Biológicos/metabolismo , Receptor de Endotelina A/metabolismo , Proteínas de Peixe-Zebra/metabolismo , Animais , Diferenciação Celular , Proliferação de Células , Receptores ErbB/antagonistas & inibidores , Melanócitos/citologia , Melanócitos/metabolismo , Melanóforos/citologia , Melanóforos/metabolismo , Modelos Biológicos , Mutação , Crista Neural/citologia , Crista Neural/metabolismo , Fenótipo , Receptor de Endotelina A/genética , Transdução de Sinais , Pigmentação da Pele/genética , Peixe-Zebra/genética , Peixe-Zebra/crescimento & desenvolvimento , Peixe-Zebra/metabolismo , Proteínas de Peixe-Zebra/antagonistas & inibidores , Proteínas de Peixe-Zebra/genética
11.
FASEB J ; 33(2): 2636-2645, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30303741

RESUMO

Adaptor protein p66Shc is overexpressed in smooth muscle cells of renal resistance vessels of hypertensive salt-sensitive rats and is involved in the regulation of renal vascular tone. We applied 2-photon laser scanning fluorescence microscopy to analyze spontaneous dynamic fluctuations in intracellular calcium concentrations ([Ca2+]i) in smooth muscle cells embedded in the walls of freshly isolated renal resistance arteries. The amplitude, number of events, and frequency of spontaneous [Ca2+]i oscillations triggered by endogenously released endothelin-1 were recorded in smooth muscle cells of the renal arteries. Endothelin receptor A antagonist BQ123 dramatically reduced the amplitude and frequency of spontaneous Ca2+ events, producing marked inhibition of renal vessels spontaneous motion. Spontaneous Ca2+ fluctuations in smooth muscle cells of p66Shc knockout (p66ShcKO) rats had significantly higher amplitude than in control rats. The frequency of spontaneous [Ca2+]i oscillations did not change in p66ShcKO rats, suggesting that p66Shc expression did not affect endothelin-1 release from resident endothelial cells. Acute application of endothelin-1 revealed significantly elevated production of the total [Ca2+]i in p66ShcKO rats. Spontaneous cytosolic Ca2+ oscillations in smooth muscle cells of renal vessels mediate their spontaneous motion via the endothelin-1/endothelin receptor A pathway. p66Shc decreases the amplitude of individual changes in [Ca2+]i, which mitigates the spontaneous motion of renal vessels.-Palygin, O., Miller, B. S., Nishijima, Y., Zhang, D. X., Staruschenko, A., Sorokin, A. Endothelin receptor A and p66Shc regulate spontaneous Ca2+ oscillations in smooth muscle cells controlling renal arterial spontaneous motion.


Assuntos
Cálcio/metabolismo , Hipertensão/fisiopatologia , Músculo Liso Vascular/fisiologia , Receptor de Endotelina A/metabolismo , Artéria Renal/fisiologia , Proteína 1 de Transformação que Contém Domínio 2 de Homologia de Src/metabolismo , Resistência Vascular , Animais , Células Cultivadas , Endotelina-1/metabolismo , Hipertensão/metabolismo , Masculino , Músculo Liso Vascular/citologia , Ratos , Ratos Endogâmicos Dahl , Artéria Renal/citologia
12.
Nat Commun ; 9(1): 5224, 2018 12 06.
Artigo em Inglês | MEDLINE | ID: mdl-30523250

RESUMO

Autoantibodies have been associated with autoimmune diseases. However, studies have identified autoantibodies in healthy donors (HD) who do not develop autoimmune disorders. Here we provide evidence of a network of immunoglobulin G (IgG) autoantibodies targeting G protein-coupled receptors (GPCR) in HD compared to patients with systemic sclerosis, Alzheimer's disease, and ovarian cancer. Sex, age and pathological conditions affect autoantibody correlation and hierarchical clustering signatures, yet many of the correlations are shared across all groups, indicating alterations to homeostasis. Furthermore, we identify relationships between autoantibodies targeting structurally and functionally related molecules, such as vascular, neuronal or chemokine receptors. Finally, autoantibodies targeting the endothelin receptor type A (EDNRA) exhibit chemotactic activity, as demonstrated by neutrophil migration toward HD-IgG in an EDNRA-dependent manner and in the direction of IgG from EDNRA-immunized mice. Our data characterizing the in vivo signatures of anti-GPCR autoantibodies thus suggest that they are a physiological part of the immune system.


Assuntos
Doença de Alzheimer/imunologia , Autoanticorpos/imunologia , Homeostase/imunologia , Neoplasias Ovarianas/imunologia , Receptores Acoplados a Proteínas-G/imunologia , Escleroderma Sistêmico/imunologia , Idoso , Sequência de Aminoácidos , Animais , Autoanticorpos/sangue , Autoanticorpos/metabolismo , Feminino , Humanos , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Imunoglobulina G/metabolismo , Masculino , Camundongos , Pessoa de Meia-Idade , Mapas de Interação de Proteínas/imunologia , Receptor de Endotelina A/genética , Receptor de Endotelina A/imunologia , Receptor de Endotelina A/metabolismo , Receptores Acoplados a Proteínas-G/genética , Receptores Acoplados a Proteínas-G/metabolismo , Homologia de Sequência de Aminoácidos
13.
Leuk Res ; 75: 61-68, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30384975

RESUMO

Endothelin receptor type A (EDNRA) is known as a mediator of cell proliferation and survival. Aberrant regulation of EDNRA has been shown to play a role in tumor growth and metastasis. Using a global gene expression screen, we found that expression of Ednra was upregulated in murine leukemia inducing cells co-expressing Hoxa9 and Meis1 compared to cells only expressing Hoxa9. The aim of this study was to explore the role of Ednra in leukemogenesis further. In a murine bone marrow transplantation model, mice transplanted with cells overexpressing Ednra and Hoxa9 succumbed to leukemia significantly earlier than mice transplanted with cells overexpressing Hoxa9 only. Furthermore, overexpression of Ednra led to increased proliferation and resistance to apoptosis of bone marrow cells in vitro. We could also show that Meis1 binds to the Ednra promoter region, suggesting a regulatory role for Meis1 in Ednra expression. Taken together, our results suggest a role for Ednra in Hoxa9/Meis1-driven leukemogenesis.


Assuntos
Regulação Leucêmica da Expressão Gênica/fisiologia , Proteínas de Homeodomínio/metabolismo , Leucemia Mieloide Aguda/metabolismo , Proteína Meis1/metabolismo , Receptor de Endotelina A/metabolismo , Animais , Humanos , Leucemia Mieloide Aguda/genética , Leucemia Mieloide Aguda/patologia , Camundongos , Camundongos Endogâmicos C57BL
14.
Invest Ophthalmol Vis Sci ; 59(12): 5256-5265, 2018 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-30383197

RESUMO

Purpose: Obstructive sleep apnea recently has been associated with a higher frequency of ischemic optic neuropathies. Intermittent hypoxia (IH) has been proposed as a major component of obstructive sleep apnea cardiovascular consequences. However, there currently are no pathophysiologic data regarding the effect of IH on the ocular vascular system. Thus, we assessed the impact of chronic IH exposure on the morphology and vascular reactivity of the rat ophthalmic artery (OA). Methods: Rats were exposed to 14 days of IH or normoxia (NX). Ophthalmic artery reactivity was studied using wire myography in rats treated or not with tempol (1 mM/day). Expression of endothelin-1 (ET-1) and its receptors, and of the three nitric oxide synthase (NOS) isoform genes was quantified using quantitative polymerase chain reaction (qPCR) in the retina and optic nerve. Structural alterations (optical and electron microscopy) and superoxide anion production were studied in OA sections. Results: Superoxide ion expression in the OA wall was increased by 23% after IH exposure. Ophthalmic artery contractile response to 3.10-8 M ET-1 was increased by 18.6% and nitric oxide-mediated relaxation was significantly delayed in IH compared to NX rats. In the absence of nitric oxide, cytochrome P450 blockade increased relaxation to acetylcholine in IH rats and delayed it in NX rats. Tempol treatment abolished the IH-induced changes in OA reactivity. Conclusions: These results strongly suggest that chronic IH induces oxidative stress in the rat OA, associated with endothelial dysfunction through alterations of nitric oxide and endothelium-derived hyperpolarising factors (EDHF) pathways.


Assuntos
Endotelina-1/metabolismo , Hipóxia/fisiopatologia , Artéria Oftálmica/fisiopatologia , Estresse Oxidativo , Receptor de Endotelina A/metabolismo , Animais , Doença Crônica , Óxidos N-Cíclicos/farmacologia , Hipóxia/metabolismo , Masculino , Músculo Liso Vascular/fisiologia , Miografia , Óxido Nítrico Sintase/genética , Artéria Oftálmica/metabolismo , Inibidores da Síntese de Proteínas/farmacologia , RNA Mensageiro/genética , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase em Tempo Real , Marcadores de Spin , Superóxidos/metabolismo
15.
Mol Med Rep ; 18(6): 5229-5236, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30272323

RESUMO

Cerebral vasospasm (CVS) is a severe complication of subarachnoid hemorrhage (SAH), and endothelin­1 (ET­1) may be involved in its pathogenesis. The present study aimed to investigate the expression of ET­1 in cerebrospinal fluid (CSF) in patients with SAH and to analyze rat arterial contractility and the expression levels of ET­1 receptors in vitro. CSF samples were collected from 28 patients and the expression levels of ET­1 were measured. Rat cerebral basilar arteries were isolated and incubated with hemorrhagic or clear CSF. Contractility, as well as ETA and ETB mRNA expression were measured. ET­1 levels in CSF increased and reached a peak within the initial 5 days after SAH onset and then gradually subsided. After 12 or 24 h, the contraction of arteries incubated in hemorrhagic CSF was substantially stronger than those in clear CSF. The mRNA expression levels of endothelin receptor type A and B in arteries incubated in hemorrhagic CSF were significantly higher than those in clear CSF. ET­1 and its receptors may be involved in the pathogenic mechanism of CVS following SAH. ET­1 expression in CSF may be used as a marker in CVS and its receptors may provide novel therapeutic targets in CVS.


Assuntos
Endotelina-1/metabolismo , Receptor de Endotelina A/metabolismo , Hemorragia Subaracnóidea/complicações , Vasoespasmo Intracraniano/etiologia , Vasoespasmo Intracraniano/fisiopatologia , Adulto , Idoso , Animais , Angiografia por Tomografia Computadorizada , Modelos Animais de Doenças , Endotelina-1/genética , Endotelina-1/farmacologia , Feminino , Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Receptor de Endotelina A/genética , Hemorragia Subaracnóidea/líquido cefalorraquidiano , Hemorragia Subaracnóidea/diagnóstico , Fatores de Tempo , Tomografia Computadorizada por Raios X
16.
Invest Ophthalmol Vis Sci ; 59(12): 5167-5175, 2018 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-30372743

RESUMO

Purpose: Endothelin-1 (ET-1) is a potent vasoconstrictor peptide implicated in retinal venous pathologies such as diabetic retinopathy and retinal vein occlusion. However, underlying mechanisms contributing to venular constriction remain unknown. Thus, we examined the roles of ET-1 receptors, extracellular calcium (Ca2+), L-type voltage-operated calcium channels (L-VOCCs), Rho kinase (ROCK), and protein kinase C (PKC) in ET-1-induced constriction of retinal venules. Methods: Porcine retinal venules were isolated and pressurized for vasoreactivity study using videomicroscopic techniques. Protein and mRNA were analyzed using molecular tools. Results: Retinal venules developed basal tone and constricted concentration-dependently to ET-1. The ETA receptor (ETAR) antagonist BQ123 abolished venular constriction to ET-1, but ETB receptor (ETBR) antagonist BQ788 had no effect on vasoconstriction. The ETBR agonist sarafotoxin S6c did not elicit vasomotor activity. In the absence of extracellular Ca2+, venules lost basal tone and ET-1-induced constriction was nearly abolished. Although L-VOCC inhibitor nifedipine also reduced basal tone and blocked vasoconstriction to L-VOCC activator Bay K8644, constriction of venules to ET-1 remained. The ROCK inhibitor H-1152 but not PKC inhibitor Gö 6983 prevented ET-1-induced vasoconstriction. Protein and mRNA expressions of ETARs and ETBRs, along with ROCK1 and ROCK2 isoforms, were detected in retinal venules. Conclusions: Extracellular Ca2+ entry via L-VOCCs is essential for developing and maintaining basal tone of porcine retinal venules. ET-1 causes significant constriction of retinal venules by activating ETARs and extracellular Ca2+ entry independent of L-VOCCs. Activation of ROCK signaling, without involvement of PKC, appears to mediate venular constriction to ET-1 in the porcine retina.


Assuntos
Cálcio/metabolismo , Endotelina-1/farmacologia , Receptor de Endotelina A/metabolismo , Veia Retiniana/fisiologia , Vasoconstrição/efeitos dos fármacos , Quinases Associadas a rho/metabolismo , Animais , Western Blotting , Canais de Cálcio Tipo L/metabolismo , Antagonistas do Receptor de Endotelina B/farmacologia , Antagonistas dos Receptores de Endotelina/farmacologia , Feminino , Masculino , Oligopeptídeos/farmacologia , Peptídeos Cíclicos/farmacologia , Piperidinas/farmacologia , Proteína Quinase C/metabolismo , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Receptor de Endotelina A/genética , Sus scrofa , Vênulas/fisiologia
17.
Sci Rep ; 8(1): 14698, 2018 10 02.
Artigo em Inglês | MEDLINE | ID: mdl-30279475

RESUMO

LAM is a rare low-grade metastasizing lung neoplasm. Inhibitors of mTOR improve clinical outcome of LAM patients by preventing loss of lung function. Nevertheless, other cell targets may be of interest for drug development. Therefore, we explored the potential role of EDN1 (endothelin) in LAM. We report an increased endothelin blood level in LAM patients as well as EDN1 overexpression and EDN1 receptor downregulation in LAM-derived primary cells and in TSC2NEG cells mutated in TSC2. We evidenced EDN pathway dysregulation based on EDN1, EDNRA, EDNRB and ARRB1 mRNA expression in LAM-derived primary cells. We showed overexpression of EDN1 and ARRB1 mRNAs in TSC2NEG cells; these cells lost their ability to respond to stimulation by endothelin. We analyzed the effects of endothelin receptor antagonists alone or in combination with rapamycin, an mTOR inhibitor, on proliferation and migration of LAM cells. Rapamycin treatment of TSC2NEG cells significantly reduced cell proliferation or migration, while none of the tested inhibitors of EDN receptors impaired these functions. We showed that TSC2NEG cells have acquired a transformed phenotype as showed by their ability to grow as spheroids in semi-solid medium and that unlike endothelin receptors antagonists, rapamycin reduced anchorage-independent cell growth and prevented expansion of TSC2NEG spheroids.


Assuntos
Endotelina-1/metabolismo , Neoplasias Pulmonares/patologia , Linfangioleiomiomatose/patologia , Esclerose Tuberosa/patologia , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Antagonistas do Receptor de Endotelina A/farmacologia , Endotelina-1/sangue , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Pulmão/patologia , Neoplasias Pulmonares/sangue , Neoplasias Pulmonares/genética , Linfangioleiomiomatose/sangue , Linfangioleiomiomatose/genética , Cultura Primária de Células , Receptor de Endotelina A/metabolismo , Receptor de Endotelina B/metabolismo , Transdução de Sinais/efeitos dos fármacos , Sirolimo/farmacologia , Esferoides Celulares , Esclerose Tuberosa/genética , Proteína 2 do Complexo Esclerose Tuberosa/genética , beta-Arrestina 1/metabolismo
18.
Clin Sci (Lond) ; 132(20): 2261-2267, 2018 10 31.
Artigo em Inglês | MEDLINE | ID: mdl-30301761

RESUMO

Cysteine-rich angiogenic inducer 61 (CYR61), an angiogenic factor whose expression is decreased in fibroids. The aim of the present study was to determine if CYR61 secretion in smooth muscle cells (SMCs) is regulated by hypoxia and through the endothelin A (ETA) receptor. SMCs from fibroids (fSMC) and the adjacent myometrium smooth muscle cells (mSMCs) were extracted from ten women undergoing hysterectomy for uterine fibroids and cultured with or without 1.0 µM of an ETA receptor antagonist for 24 h under either normal or hypoxic oxygen conditions. Cellular secretion of endothelin-1 (ET-1) and CYR61 were measured via enzyme linked immunosorbent assay in the cell culture media. SMCs were collected to determine cell proliferation and CYR61 protein expression via Western blot. ET-1 secretion was significantly increased in fSMC and was decreased with blockade of the ETA receptor under both normoxia (P=0.0004) and hypoxia (P=0.008). CYR61 expression was decreased in fSMCs and significantly increased with blockade of the ETA receptor under hypoxia (P=0.04). Cell proliferation decreased with ETA blockade under normoxia (P=0.0001) and hypoxia (P=0.001). These results suggest that suppression of CYR61 secretion in fSMC is regulated by the ET-1 and that blockade with ETA could be considered for a future treatment option.


Assuntos
Leiomioma/cirurgia , Miócitos de Músculo Liso/metabolismo , Receptor de Endotelina A/metabolismo , Células Cultivadas , Proteína Rica em Cisteína 61/metabolismo , Antagonistas do Receptor de Endotelina A/farmacologia , Feminino , Humanos , Histerectomia/métodos , Leiomioma/metabolismo , Miócitos de Músculo Liso/efeitos dos fármacos , Miométrio/metabolismo , Miométrio/patologia , Oxigênio/metabolismo , Neoplasias Uterinas/metabolismo , Neoplasias Uterinas/cirurgia
19.
Physiol Rep ; 6(17): e13865, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-30198212

RESUMO

Renal ischemia-reperfusion (IR) injury and acute kidney injury (AKI) increase the risk of developing hypertension, with T cells suspected as a possible mechanistic link. Endothelin promotes renal T cell infiltration in several diseases, predominantly via the ETA receptor, but its contribution to renal T cell infiltration following renal IR injury is poorly understood. To test whether ETA receptor activation promotes T cell infiltration of the kidney following IR injury, male C57BL/6 mice were treated with the ETA receptor antagonist ABT-627 or vehicle, commencing 2 days prior to unilateral renal IR injury. Mice were sacrificed at 24 h or 10 days post-IR for assessment of the initial renal injury and subsequent infiltration of T cells. Vehicle and ABT-627-treated mice displayed significant upregulation of endothelin-1 (ET-1) in the IR compared to contralateral kidney at both 24 h and 10 days post-IR (P < 0.001). Renal CD3+ T cell numbers were increased in the IR compared to contralateral kidneys at 10 days, but ABT-627-treated mice displayed a 35% reduction in this effect in the outer medulla (P < 0.05 vs. vehicle) and a nonsignificant 23% reduction in the cortex compared to vehicle-treated mice. Whether specific T cell subsets were affected awaits confirmation by flow cytometry, but outer medullary expression of the T helper 17 transcription factor RORγt was reduced by ABT-627 (P = 0.06). These data indicate that ET-1 acting via the ETA receptor contributes to renal T cell infiltration post-IR injury. This may have important implications for immune system-mediated long-term consequences of AKI, an area which awaits further investigation.


Assuntos
Rim/metabolismo , Receptor de Endotelina A/metabolismo , Traumatismo por Reperfusão/metabolismo , Linfócitos T/fisiologia , Animais , Movimento Celular , Antagonistas do Receptor de Endotelina A/farmacologia , Endotelina-1/metabolismo , Rim/irrigação sanguínea , Rim/patologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Membro 3 do Grupo F da Subfamília 1 de Receptores Nucleares/metabolismo , Traumatismo por Reperfusão/patologia
20.
Brain Res ; 1701: 196-203, 2018 12 15.
Artigo em Inglês | MEDLINE | ID: mdl-30244111

RESUMO

BACKGROUND: Cerebral vasospasm may lead to delayed ischemic neurological deficits following subarachnoid hemorrhage (SAH). Endothelin (ET-1) is an important factor participating in cerebral vasospasm underlying SAH. We used a specific endothelin receptor antagonist, BQ123 to assess the specific role of endothelin-1 receptor antagonist in cerebral vasospasm in a rabbit model of SAH by examining plasma ET-1 levels and the principal CT perfusion (CTP) parameters pertinent to the hemodynamic status of microcirculation following SAH. METHODS: 102 male New Zealand white rabbits were divided into control, SAH and SAH + BQ123 intervention group (BQ123 group). Rabbit SAH model was established by double hemorrhage injection of autologous blood into the cisterna magna; Aquilion ONE was used to collect cerebral blood flow (CBF), cerebral blood volume (CBV), and mean transit time (MTT) which were used to evaluate cerebral microcirculation hemodynamics; Elisa was used to assess plasma ET-1 levels. Data were collected on days 1, 4, 7 and 14 following SAH, respectively. RESULTS: Compared with the control group, the CBF in the SAH group was significantly lower, while the MTT was significantly higher. The CBF decreased on the 4th day and reached the lowest on the 7th day. The MTT began to rise on the 4th day and peaked on the 7th day. While in the BQ123 intervention group, the CBF significantly increased while the MTT significantly decreased on the 1st and the 4th days, respectively. Compared with SAH group, plasma ET-1 levels in BQ123 group significantly increased on the earlier (1st and 4th days) but not later days (between the 7th and 14th days). In addition, the inflammatory infiltration of brain tissues in rabbits treated with BQ123 post-SAH was significantly reduced compared with SAH group. CONCLUSION: CTP can quantify the therapeutic effect of BQ123 after SAH; Selective blockade of ET-1 endothelin receptor, BQ123 significantly improved microcirculatory perfusion along with a reduction in resultant vasogenic inflammatory responses. The effect of BQ123 on the cerebral microcirculation was lobe dependent.


Assuntos
Antagonistas do Receptor de Endotelina A/metabolismo , Peptídeos Cíclicos/farmacologia , Hemorragia Subaracnóidea/fisiopatologia , Animais , Circulação Cerebrovascular/fisiologia , Endotelina-1/sangue , Endotelina-1/metabolismo , Endotelinas , Hemodinâmica , Masculino , Microcirculação/fisiologia , Peptídeos Cíclicos/metabolismo , Imagem de Perfusão , Coelhos , Receptor de Endotelina A/metabolismo , Hemorragia Subaracnóidea/diagnóstico por imagem , Tomografia Computadorizada por Raios X/métodos , Vasoespasmo Intracraniano
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA