Asunto(s)
Receptores de Activinas Tipo II/sangre , Malformaciones Arteriovenosas/etiología , Proteínas Morfogenéticas Óseas/sangre , Procedimiento de Fontan , Factor 2 de Diferenciación de Crecimiento/sangre , Síndrome del Corazón Izquierdo Hipoplásico/cirugía , Complicaciones Posoperatorias/etiología , Adolescente , Malformaciones Arteriovenosas/sangre , Malformaciones Arteriovenosas/prevención & control , Biomarcadores/sangre , Estudios de Casos y Controles , Niño , Preescolar , Femenino , Humanos , Síndrome del Corazón Izquierdo Hipoplásico/sangre , Síndrome del Corazón Izquierdo Hipoplásico/fisiopatología , Lactante , Masculino , Complicaciones Posoperatorias/sangre , Complicaciones Posoperatorias/prevención & control , Arteria Pulmonar/anomalías , Venas Pulmonares/anomalíasRESUMEN
Aims: Hereditary hemorrhagic telangiectasia is an autosomal dominant disorder characterized by telangiectasia, epistaxis, and vascular malformations. Pathogenic mutations were found in ENG, AVCRL1, SMAD4, and GDF genes. In this study, we present our database of patients with hereditary hemorrhagic telangiectasia regarding the phenotype-genotype relations and discuss two novel ENG gene pathogenic variations in two unrelated families. Methods: Next Generation Sequencing analysis was performed on the peripheral blood of nine patients with hereditary hemorrhagic telangiectasia in four unrelated families. All patients were diagnosed with hereditary hemorrhagic telangiectasia according to the Curaçao criteria. Data on treatment and screenings of visceral involvement were recorded from files. Results: We have found a pathogenic variation in either the ENG or ACVRL1 gene in each family. Two novel pathogenic variations in the ENG gene, including NM_000118.3 (ENG): c.416delC (p.P139fs*24) and NM_000118.3(ENG): c.1139dupT (p.Leu380PhefsTer16), were found in the same family. The NM_000020.2(ACVRL1): c.1298C>T (p.Pro433Leu) pathogenic variation in the ACVRL1 gene in our first family and a novel heterozygous likely pathogenic NM_000020.2(ACVRL1): c.95T>C (p.Val32Ala) variation was found in our second family. Seven of the nine patients were treated with thalidomide for controlling bleeding episodes. All patients responded to thalidomide. In one patient, the response to thalidomide was lost and switched to bevacizumab. Conclusion: In HHT certain type of mutations correlates with disease phenotypes and with next generation sequencing method, new pathogenic variations can be revealed which might help managing HHT patients.
Asunto(s)
Telangiectasia Hemorrágica Hereditaria/sangre , Factores de Virulencia , Receptores de Activinas Tipo II/análisis , Receptores de Activinas Tipo II/sangre , Adulto , Anciano , Endoglina/análisis , Endoglina/sangre , Femenino , Humanos , Masculino , Persona de Mediana Edad , Proteína Smad4/análisis , Proteína Smad4/sangre , Telangiectasia Hemorrágica Hereditaria/fisiopatología , TurquíaRESUMEN
The ability to safely control transgene expression with simple synthetic gene switches is critical for effective gene- and cell-based therapies. In the present study, the signaling pathway controlled by human transient receptor potential (TRP) melastatin 8 (hTRPM8), a TRP channel family member1, is harnessed to control transgene expression. Human TRPM8 signaling is stimulated by menthol, an innocuous, natural, cooling compound, or by exposure to a cool environment (15-18 °C). By functionally linking hTRPM8-induced signaling to a synthetic promoter containing elements that bind nuclear factor of activated T cells, a synthetic gene circuit was designed that can be adjusted by exposure to either a cool environment or menthol. It was shown that this gene switch is functional in various cell types and human primary cells, as well as in mice implanted with engineered cells. In response to transdermal delivery of menthol, microencapsulated cell implants harboring this gene circuit, coupled to expression of either of two therapeutic proteins, insulin or a modified, activin type IIB, receptor ligand trap protein (mActRIIBECD-hFc), could alleviate hyperglycemia in alloxan-treated mice (a model of type 1 diabetes) or reverse muscle atrophy in dexamethasone-treated mice (a model of muscle wasting), respectively. This fully human-derived orthogonal transgene switch should be amenable to a wide range of clinical applications.
Asunto(s)
Receptores de Activinas Tipo II/sangre , Insulina/biosíntesis , Canales Catiónicos TRPM/fisiología , Transgenes , Receptores de Activinas Tipo II/genética , Receptores de Activinas Tipo II/uso terapéutico , Fosfatasa Alcalina/genética , Animales , Frío , Diabetes Mellitus Tipo 1/tratamiento farmacológico , Sistemas de Liberación de Medicamentos , Femenino , Regulación de la Expresión Génica , Células HEK293 , Humanos , Insulina/genética , Insulina/uso terapéutico , Mentol/farmacología , Ratones , Ratones Endogámicos C57BL , Distrofias Musculares/tratamiento farmacológico , Transducción de Señal/efectos de los fármacosRESUMEN
Background: Preeclampsia (PE) is the most common complication of pregnancy that remains to be a major cause of maternal and fetal mortality. Prediction and early diagnosis of PE would allow for timely initiation of preventive therapy. According to recent studies of ACVR2A gene polymorphism is associated with PE, but it is still unclear whether these findings reflect specific pathogenetic mechanisms of this disease. Methods: We performed targeted next-generation sequencing (NGS) sequencing of ACVR2A gene by means of Ion Torrent Personal Genome machine (PGM) Sequencer. A genetic analysis of patients with PE and control group was performed. Bioinformatics analysis using Polyphen2 (Boston, MA), SIFT (La Jolla, CA), and SnpSift software were used. To select genetic markers in PE patients two additive models and score analysis were applied. Results: Based on the score analysis, we detected two substitutions (rs145399059 and rs17692648) and one insertion insAA at position 148642724 that were associated with PE in our cohorts. We also detected a variant rs17742573 that can be considered as protective against preeclampsia. Conclusions: Our data suggest that some variants in ACVR2A gene are associated with PE. But more studies are required to reveal the role of ACVR2A gene in the pathogenesis of this disease during pregnancy.
Asunto(s)
Receptores de Activinas Tipo II/genética , Preeclampsia/genética , Receptores de Activinas Tipo II/sangre , Adulto , Biomarcadores , Estudios de Casos y Controles , Femenino , Marcadores Genéticos , Predisposición Genética a la Enfermedad , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Polimorfismo de Nucleótido Simple , Preeclampsia/diagnóstico , Embarazo , Resultado del EmbarazoRESUMEN
Activin A, a multifunctional cytokine of transforming growth factor-ß (TGF-ß) superfamily, can be produced by the diverse immune cells. NK cells in peripheral blood are one of the major immune cells applied to cancer therapy in recent years. However, whether activin A can be produced by natural killer (NK) cells and be involved in regulation of peripheral blood NK cells activities of mouse are not well characterized. Here, we found that activin type IIA and IIB receptors and signaling molecules Smad2, 3 were expressed in peripheral blood NK cells of mouse by flow cytometry and RT-PCR. The cultured blood NK cells of mouse not only produced activin ßA chain protein by intracellular cytokine staining, but also secreted mature activin A protein by enzyme-linked immunosorbent assay (ELISA), and the production was promoted by IL-2. In addition, IL-2 as a positive control obviously promoted IFNγ production of mouse blood NK cells in vitro. However, activin A suppressed IFNγ production, but enhanced IL-2 synthesis and did not alter IL-10 production. Moreover, we found that activin A significantly suppressed the ability of NK cells to lyse target cells. These data revealed that blood NK cells of mouse were not only the target cells in response to activin A, but also the source of activin A, suggesting that activin A may play an important role in regulation of NK cells activities of mouse in an autocrine / paracrine manner.
Asunto(s)
Activinas/farmacología , Comunicación Autocrina , Células Asesinas Naturales/metabolismo , Comunicación Paracrina , Receptores de Activinas Tipo II/sangre , Receptores de Activinas Tipo II/genética , Receptores de Activinas Tipo II/metabolismo , Animales , Comunicación Autocrina/efectos de los fármacos , Muerte Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Folistatina/farmacología , Subunidades beta de Inhibinas/sangre , Subunidades beta de Inhibinas/genética , Subunidades beta de Inhibinas/metabolismo , Interferón gamma/biosíntesis , Interleucina-10/biosíntesis , Interleucina-2/biosíntesis , Células Asesinas Naturales/efectos de los fármacos , Masculino , Ratones Endogámicos BALB C , Comunicación Paracrina/efectos de los fármacos , ARN Mensajero/genética , ARN Mensajero/metabolismo , Proteínas Smad/sangre , Proteínas Smad/genética , Proteínas Smad/metabolismoRESUMEN
PURPOSE: Inhibitors of the ActRII signaling pathways represent promising therapeutics for the treatment of muscular diseases, but also pose risks as performance-enhancing agents in sports. Bimagrumab is a human anti-ActRII antibody which was found to increase muscle mass and function by blocking ActRII signaling. As it has considerable potential for being misused as doping agent in sports, the aim of this study was to develop a mass spectrometric detection assay for doping control serum samples. EXPERIMENTAL DESIGN: Within this study, a detection method for Bimagrumab in human serum was developed, which combines ammonium sulfate precipitation and affinity purification with proteolytic digestion and LC-HRMS. To facilitate the unambiguous identification of the diagnostic peptides, an orthogonal IM separation was additionally performed. RESULTS: The assay was successfully validated and the analysis of clinical samples demonstrated its fitness for purpose for an application in routine doping control analysis. CONCLUSIONS AND CLINICAL RELEVANCE: Although no myostatin inhibitors have obtained clinical approval yet, the proactive development of detection methods for emerging doping agents represents a key aspect of preventive doping research. The presented approach will expand the range of available tests for novel protein therapeutics and can readily be modified to include further target analytes.
Asunto(s)
Receptores de Activinas Tipo II/sangre , Anticuerpos Bloqueadores/sangre , Anticuerpos Bloqueadores/inmunología , Anticuerpos Monoclonales/sangre , Anticuerpos Monoclonales/inmunología , Análisis Químico de la Sangre/métodos , Proteolisis , Tripsina/metabolismo , Receptores de Activinas Tipo II/inmunología , Secuencia de Aminoácidos , Anticuerpos Bloqueadores/química , Anticuerpos Monoclonales/química , Anticuerpos Monoclonales Humanizados , Cromatografía de Afinidad , Cromatografía Liquida , Humanos , Masculino , Espectrometría de MasasRESUMEN
OBJECTIVES: Activin A, a TGFß family member, circulates in the maternal blood at increasing concentrations throughout gestation during a healthy pregnancy. The circulating concentration of activin A is further increased in pre-eclampsia (PE), a hypertensive disorder of pregnancy that is marked by systemic maternal vascular endothelial cell dysfunction. The effect of increasing activin A concentrations on the maternal vascular endothelium is unknown. The study aim was to investigate the effect of physiological and pathological activin A concentrations observed in normotensive and PE pregnancies respectively, on vascular endothelial cell function. METHODS AND RESULTS: Immunostaining demonstrated the presence of the activin A receptor, ACVR2A, in SGHEC-7 cells used to model the vascular endothelium. SGHEC-7 cells were treated with activin A concentrations representative of concentrations throughout gestation in normotensive (0-10ng/ml) and PE (50ng/ml) pregnancies. xCELLigence functional assays revealed that normotensive activin A concentrations increased SGHEC-7 proliferation and migration, which was inhibited by PE concentrations. Additionally, fluorescence based assays showed that PE concentrations increased endothelial permeability. None of the tested activin A concentrations affected cell apoptosis. PE concentrations also resulted in an imbalance of the vasoactive factors eNOS, PTGIS and EDN1, as determined by quantitative real-time polymerase chain reaction and enzyme-linked immunosorbent assays. CONCLUSION: Compared with normotensive activin A concentrations, the higher PE activin A concentrations resulted in abnormal endothelial functions, which may contribute to the systemic maternal vascular endothelial cell dysfunction observed in the disorder.
Asunto(s)
Activinas/sangre , Endotelio Vascular , Preeclampsia/sangre , Preeclampsia/diagnóstico , Receptor 1 de Factores de Crecimiento Endotelial Vascular/sangre , Receptores de Activinas Tipo II/sangre , Adulto , Biomarcadores/sangre , Técnicas de Cultivo de Célula , Endotelio Vascular/fisiopatología , Femenino , Humanos , Preeclampsia/fisiopatología , Valor Predictivo de las Pruebas , Embarazo , Sensibilidad y EspecificidadRESUMEN
Diamond-Blackfan Anemia (DBA) is a bone marrow failure disorder characterized by low red blood cell count. Mutations in ribosomal protein genes have been identified in approximately half of all DBA cases. Corticosteriod therapy and bone marrow transplantation are common treatment options for patients; however, significant risks and complications are associated with these treatment options. Therefore, novel therapeutic approaches are needed for treating DBA. Sotatercept (ACE-011, and its murine ortholog RAP-011) acts as an activin receptor type IIA ligand trap, increasing hemoglobin and hematocrit in pharmacologic models, in healthy volunteers, and in patients with ß-thalassemia, by expanding late-stage erythroblasts through a mechanism distinct from erythropoietin. Here, we evaluated the effects of RAP-011 in zebrafish models of RPL11 ribosome deficiency. Treatment with RAP-011 dramatically restored hemoglobin levels caused by ribosome stress. In zebrafish embryos, RAP-011 likely stimulates erythropoietic activity by sequestering lefty1 from erythroid cells. These findings identify lefty1 as a signaling component in the development of erythroid cells and rationalize the use of sotatercept in DBA patients.