RESUMEN
Somatic inactivating mutations in epigenetic regulators are frequently found in combination in myelodysplastic syndrome (MDS). However, the mechanisms by which combinatory mutations in epigenetic regulators promote the development of MDS remain unknown. Here we performed epigenomic profiling of hematopoietic progenitors in MDS mice hypomorphic for Tet2 following the loss of the polycomb-group gene Ezh2 (Tet2KD/KDEzh2Δ/Δ). Aberrant DNA methylation propagated in a sequential manner from a Tet2-insufficient state to advanced MDS with deletion of Ezh2. Hyper-differentially methylated regions (hyper-DMRs) in Tet2KD/KDEzh2Δ/Δ MDS hematopoietic stem/progenitor cells were largely distinct from those in each single mutant and correlated with transcriptional repression. Although Tet2 hypomorph was responsible for enhancer hypermethylation, the loss of Ezh2 induced hyper-DMRs that were enriched for CpG islands of polycomb targets. Notably, Ezh2 targets largely lost the H3K27me3 mark while acquiring a significantly higher level of DNA methylation than Ezh1 targets that retained the mark. These findings indicate that Ezh2 targets are the major targets of the epigenetic switch in MDS with Ezh2 insufficiency. Our results provide a detailed trail for the epigenetic drift in a well-defined MDS model and demonstrate that the combined dysfunction of epigenetic regulators cooperatively remodels the epigenome in the pathogenesis of MDS.
Asunto(s)
Proteínas de Unión al ADN/metabolismo , Proteína Potenciadora del Homólogo Zeste 2/metabolismo , Epigénesis Genética , Regulación de la Expresión Génica , Síndromes Mielodisplásicos/genética , Síndromes Mielodisplásicos/metabolismo , Proteínas Proto-Oncogénicas/metabolismo , Animales , Secuencia de Bases , Sitios de Unión , Islas de CpG , Metilación de ADN , Proteínas de Unión al ADN/genética , Dioxigenasas , Modelos Animales de Enfermedad , Elementos de Facilitación Genéticos , Proteína Potenciadora del Homólogo Zeste 2/genética , Hematopoyesis/genética , Ratones , Ratones Noqueados , Ratones Transgénicos , Miembro 2 del Grupo A de la Subfamilia 4 de Receptores Nucleares/genética , Miembro 2 del Grupo A de la Subfamilia 4 de Receptores Nucleares/metabolismo , Motivos de Nucleótidos , Unión Proteica , Proteínas Proto-Oncogénicas/genética , Proteínas Represoras/metabolismo , Factores de Transcripción/metabolismoRESUMEN
Mutations of calreticulin (CALR) are detected in 25-30% of patients with essential thrombocythemia (ET) or primary myelofibrosis and cause frameshifts that result in proteins with a novel C-terminal. We demonstrate that CALR mutations activated signal transducer and activator of transcription 5 (STAT5) in 293T cells in the presence of thrombopoietin receptor (MPL). Human megakaryocytic CMK11-5 cells and erythroleukemic F-36P-MPL cells with knocked-in CALR mutations showed increased growth and acquisition of cytokine-independent growth, respectively, accompanied by STAT5 phosphorylation. Transgenic mice expressing a human CALR mutation with a 52 bp deletion (CALRdel52-transgenic mice (TG)) developed ET, with an increase in platelet count, but not hemoglobin level or white blood cell count, in association with an increase in bone marrow (BM) mature megakaryocytes. CALRdel52 BM cells did not drive away wild-type (WT) BM cells in in vivo competitive serial transplantation assays, suggesting that the self-renewal capacity of CALRdel52 hematopoietic stem cells (HSCs) was comparable to that of WT HSCs. Therapy with the Janus kinase (JAK) inhibitor ruxolitinib ameliorated the thrombocytosis in TG mice and attenuated the increase in number of BM megakaryocytes and HSCs. Taken together, our study provides a model showing that the C-terminal of mutant CALR activated JAK-STAT signaling specifically downstream of MPL and may have a central role in CALR-induced myeloproliferative neoplasms.
Asunto(s)
Calreticulina/genética , Animales , Autorrenovación de las Células , Células HEK293 , Células Madre Hematopoyéticas , Humanos , Quinasas Janus/antagonistas & inhibidores , Ratones , Ratones Transgénicos , Trastornos Mieloproliferativos/inducido químicamente , Trastornos Mieloproliferativos/etiología , Nitrilos , Inhibidores de Proteínas Quinasas/farmacología , Inhibidores de Proteínas Quinasas/uso terapéutico , Pirazoles/farmacología , Pirazoles/uso terapéutico , Pirimidinas , Receptores de Trombopoyetina , Factor de Transcripción STAT5/metabolismo , Trombocitemia Esencial/tratamiento farmacológico , Trombocitemia Esencial/genéticaAsunto(s)
Antidepresivos Tricíclicos , Citocromo P-450 CYP2C19/genética , Citocromo P-450 CYP2D6/genética , Técnicas de Genotipaje/métodos , Administración del Tratamiento Farmacológico/normas , Variantes Farmacogenómicas/genética , Antidepresivos Tricíclicos/efectos adversos , Antidepresivos Tricíclicos/farmacocinética , Relación Dosis-Respuesta a Droga , Humanos , Polimorfismo Genético , Resultado del TratamientoRESUMEN
Joubert syndrome (JS) is rare recessive disorders characterized by the combination of hypoplasia/aplasia of the cerebellar vermis, thickened and elongated superior cerebellar peduncles, and a deep interpeduncular fossa which is defined by neuroimaging and is termed the 'molar tooth sign'. JS is genetically highly heterogeneous, with at least 29 disease genes being involved. To further understand the genetic causes of JS, we performed whole-exome sequencing in 24 newly recruited JS families. Together with six previously reported families, we identified causative mutations in 25 out of 30 (24 + 6) families (83.3%). We identified eight mutated genes in 27 (21 + 6) Japanese families, TMEM67 (7/27, 25.9%) and CEP290 (6/27, 22.2%) were the most commonly mutated. Interestingly, 9 of 12 CEP290 disease alleles were c.6012-12T>A (75.0%), an allele that has not been reported in non-Japanese populations. Therefore c.6012-12T>A is a common allele in the Japanese population. Importantly, one Japanese and one Omani families carried compound biallelic mutations in two distinct genes (TMEM67/RPGRIP1L and TMEM138/BBS1, respectively). BBS1 is the causative gene in Bardet-Biedl syndrome. These concomitant mutations led to severe and/or complex clinical features in the patients, suggesting combined effects of different mutant genes.
Asunto(s)
Anomalías Múltiples/genética , Proteínas Adaptadoras Transductoras de Señales/genética , Antígenos de Neoplasias/genética , Cerebelo/anomalías , Anomalías del Ojo/genética , Enfermedades Renales Quísticas/genética , Proteínas de la Membrana/genética , Proteínas Asociadas a Microtúbulos/genética , Proteínas de Neoplasias/genética , Retina/anomalías , Anomalías Múltiples/diagnóstico por imagen , Anomalías Múltiples/epidemiología , Anomalías Múltiples/fisiopatología , Alelos , Proteínas de Ciclo Celular , Cerebelo/diagnóstico por imagen , Cerebelo/fisiopatología , Proteínas del Citoesqueleto , Anomalías del Ojo/diagnóstico por imagen , Anomalías del Ojo/epidemiología , Anomalías del Ojo/fisiopatología , Femenino , Heterogeneidad Genética , Predisposición Genética a la Enfermedad , Humanos , Japón/epidemiología , Enfermedades Renales Quísticas/diagnóstico por imagen , Enfermedades Renales Quísticas/epidemiología , Enfermedades Renales Quísticas/fisiopatología , Masculino , Mutación , Omán/epidemiología , Linaje , Retina/diagnóstico por imagen , Retina/fisiopatologíaAsunto(s)
ADP-Ribosil Ciclasa 1/metabolismo , Citotoxicidad Inmunológica/efectos de los fármacos , Interferón-alfa/farmacología , Leucemia-Linfoma de Células T del Adulto/inmunología , Glicoproteínas de Membrana/metabolismo , Linfocitos T/efectos de los fármacos , Tretinoina/farmacología , ADP-Ribosil Ciclasa 1/inmunología , Adulto , Sinergismo Farmacológico , Humanos , Interferón-alfa/administración & dosificación , Leucemia-Linfoma de Células T del Adulto/patología , Glicoproteínas de Membrana/inmunología , Receptores de Antígenos de Linfocitos T/inmunología , Receptores de Antígenos de Linfocitos T/metabolismo , Proteínas Recombinantes de Fusión/inmunología , Proteínas Recombinantes de Fusión/metabolismo , Linfocitos T/inmunología , Linfocitos T/metabolismo , Tretinoina/administración & dosificación , Células Tumorales Cultivadas , Regulación hacia Arriba/efectos de los fármacosRESUMEN
INTRODUCTION: This study evaluated the effects of the CYP2D6*10 genotype on steady-state plasma concentrations of enantiomeric mirtazapine (MIR) and N-desmethylmirtazapine (DMIR) in Japanese patients. METHODS: Subjects were 77 Japanese patients treated with racemic MIR. Steady-state plasma concentrations of MIR and DMIR enantiomers were measured using stereoselective liquid chromatography. Polymerase chain reaction was used to determine the CYP2D6 genotypes. RESULTS: After correcting for dose and body weight, smokers (n=15) had significantly lower S-(+)-MIR than nonsmokers (n=55) (15.1±17.8 vs. 23.9±17.8 ng/mL/mg/kg, Kruskal-Wallis test, p=0.034). One-way analysis of variance revealed that CYP2D6*10 homozygotes had significantly higher corrected plasma concentrations of S-(+)-MIR than the no-variant allele group (p=0.034). Multiple regression analysis revealed a significant positive correlation between the number of CYP2D6*10 alleles and corrected plasma concentrations of S-(+)-MIR. These results yielded the following final model: corrected plasma concentration of S-(+)-MIR=15.9+7.30×(number of CYP2D6*10 alleles) (R=0.279, p=0.023, coefficient of determination (R(2))=0.078). CONCLUSION: Homozygous CYP2D6*10 alleles and smoking have a significant impact on the metabolism of S-(+)-MIR in Japanese patients.
Asunto(s)
Antidepresivos Tricíclicos/sangre , Citocromo P-450 CYP2D6/genética , Trastorno Depresivo/tratamiento farmacológico , Genotipo , Mianserina/análogos & derivados , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Alelos , Antidepresivos Tricíclicos/uso terapéutico , Trastorno Depresivo/sangre , Trastorno Depresivo/genética , Femenino , Humanos , Japón , Masculino , Mianserina/sangre , Mianserina/uso terapéutico , Persona de Mediana Edad , Mirtazapina , Farmacogenética , Adulto JovenRESUMEN
A single somatic mutation, V617F, in Janus kinase 2 (JAK2) is one of the causes of myeloproliferative neoplasms (MPNs), including primary myelofibrosis, and the JAK2V617F mutant kinase is a therapeutic target in MPN. However, inhibition of wild-type (WT) JAK2 can decrease the erythrocyte or platelet (PLT) count. Our selective JAK2 inhibitor, NS-018, suppressed the growth of Ba/F3 cells harboring JAK2V617F more strongly than that of cells harboring WT JAK2. The 4.3-fold JAK2V617F selectivity of NS-018 is higher than the 1.0- to 2.9-fold selectivity of seven existing JAK2 inhibitors. NS-018 also inhibited erythroid colony formation in JAK2V617F transgenic mice at significantly lower concentrations than in WT mice. In keeping with the above results, in a JAK2V617F bone marrow transplantation mouse model with a myelofibrosis-like disease, NS-018 reduced leukocytosis and splenomegaly, improved bone marrow fibrosis and prolonged survival without decreasing the erythrocyte or PLT count in the peripheral blood. By exploring the X-ray co-crystal structure of NS-018 bound to JAK2, we identified unique hydrogen-bonding interactions between NS-018 and Gly993 as a plausible explanation for its JAK2V617F selectivity. These results suggest that NS-018 will have therapeutic benefit for MPN patients through both its efficacy and its reduced hematologic adverse effects.
RESUMEN
In the larval bester, a hybrid sturgeon of beluga Huso huso and sterlet Acipenser ruthenus, development of cartilage around the notochord began 7 days post hatch (dph) (14.0 mm, total length, L(T)). The vertebral cartilage develops in the following sequence: basidorsals and basiventrals, neural canals, neural spines and ribs. The development of ribs remained incomplete in the largest specimen (181 dph, 179 mm L(T) ) that was examined. Endoskeletal development of the fins began 4 dph for the dorsal and anal fins, 6 dph for the pectoral fin and 10 dph for the caudal and pelvic fins. Complete elements of all fins were observed by 91 dph and complete ossification of fin rays was observed by 122 dph in the double-stained specimens. Observation of the histological sections, however, suggested that ossification occurred soon after the formation of the organic matrix in the fin rays. Dorsal scutes were first visible by 25 dph, followed by the lateral and ventral scutes, which were visible by 37 and 44 dph, respectively. The number of scutes was fixed at 44, 59 and 91 dph and ossification was complete by 59 (dorsal) and 91 dph (lateral and ventral scutes) in the double-stained specimens. Ossification occurred soon after the formation of the scute organic matrix in the histological sections. Four types of scales were observed in the H. huso×A. ruthenus hybrid. Median predorsal, preanal and small scales on the anterior section of the head were visible by 59 dph. Scales on the caudal fin were visible by 91 dph and a variable assemblage of scales anterior to the anal fin was visible by 122 dph. Both the scutes and scales developed in a process that is similar to that of intramembranous ossification.
Asunto(s)
Aletas de Animales/crecimiento & desarrollo , Peces/crecimiento & desarrollo , Columna Vertebral/crecimiento & desarrollo , Animales , Cartílago/crecimiento & desarrollo , Larva/crecimiento & desarrolloRESUMEN
An increased incidence of sudden death has been observed among patients treated with antidepressants. A prolonged QTc interval is a known prognostic factor for fatal arrhythmia, and several studies have shown that the use of antidepressants can cause a prolonged QTc interval. However, few studies, especially in Japan, have compared the effects of multiple drugs on QTc interval or examined dose relationships in a clinical setting.We compared the effects of antidepressants on QT interval, corrected to QTc by Bazett's formula, in 729 Japanese patients who were diagnosed with mood disorder.Using stepwise multiple linear regression analysis, we found that the use of tricyclic antidepressants (P<0.01) and concomitant use of antipsychotics (P<0.05), as well as advanced age and being female (known factors for prolonged QTc interval; both P<0.01), significantly prolonged the QTc interval. Analysis of individual antidepressants also revealed that the use of clomipramine (P<0.01) and amitriptyline (P<0.05) significantly prolonged the QTc interval.Our results reveal that tricyclic antidepressants, especially clomipramine and amitriptyline, confer a risk of prolonged QTc interval in a dose-dependent manner. The selective serotonin reuptake inhibitors investigated (fluvoxamine, paroxetine, sertraline) were not indicated as risk factors for QTc prolongation.
Asunto(s)
Antidepresivos Tricíclicos/efectos adversos , Antipsicóticos/efectos adversos , Pueblo Asiatico/psicología , Síndrome de QT Prolongado/inducido químicamente , Trastornos del Humor/fisiopatología , Inhibidores Selectivos de la Recaptación de Serotonina/efectos adversos , Factores de Edad , Antidepresivos Tricíclicos/administración & dosificación , Antipsicóticos/administración & dosificación , Relación Dosis-Respuesta a Droga , Quimioterapia Combinada/efectos adversos , Electrocardiografía/psicología , Electrocardiografía/estadística & datos numéricos , Femenino , Humanos , Masculino , Persona de Mediana Edad , Trastornos del Humor/tratamiento farmacológico , Análisis de Regresión , Inhibidores Selectivos de la Recaptación de Serotonina/administración & dosificación , Caracteres SexualesRESUMEN
Ten-Eleven-Translocation 2 (TET2) is an enzyme that catalyzes the conversion of 5-methylcytosine into 5-hydroxymethylcytosine (5-hmC) and thereby alters the epigenetic state of DNA; somatic loss-of-function mutations of TET2 are frequently observed in patients with diverse myeloid malignancies. To study the function of TET2 in vivo, we analyzed Ayu17-449 (TET2(trap)) mice, in which a gene trap insertion in intron 2 of TET2 reduces TET2 mRNA levels to about 20% of that found in wild-type (WT) mice. TET2(trap/trap) mice were born at Mendelian frequency but died at a high rate by postnatal day 3, indicating the essential role of TET2 for survival. Loss of TET2 results in an increase in the number of hematopoietic stem cells (HSCs)/progenitors in the fetal liver, and TET2(trap/trap) HSCs exhibit an increased self-renewal ability in vivo. In competitive transplantation assays, TET2(trap/trap) HSCs possess a competitive growth advantage over WT HSCs. These data indicate that TET2 has a critical role in survival and HSC homeostasis.
Asunto(s)
Proteínas de Unión al ADN/fisiología , Células Madre Hematopoyéticas/fisiología , Homeostasis , Proteínas Proto-Oncogénicas/fisiología , Animales , Supervivencia Celular , Dioxigenasas , Hematopoyesis , Células Madre Hematopoyéticas/citología , Janus Quinasa 2/fisiología , Ratones , Ratones Endogámicos C57BL , ARN Mensajero/análisisAsunto(s)
Médula Ósea/metabolismo , Subunidad alfa del Factor 1 Inducible por Hipoxia/genética , Leucemia Mieloide Aguda/genética , Leucemia Mieloide Aguda/patología , Células Madre Neoplásicas/metabolismo , Animales , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Médula Ósea/patología , Huesos/metabolismo , Huesos/patología , Hipoxia de la Célula/genética , Hipoxia de la Célula/fisiología , Línea Celular Tumoral , Regulación Leucémica de la Expresión Génica , Células HL-60 , Humanos , Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismo , Células K562 , Leucemia Mieloide Aguda/tratamiento farmacológico , Leucemia Mieloide Aguda/metabolismo , Ratones , Neoplasia Residual/etiología , Neoplasia Residual/genética , Neoplasia Residual/metabolismo , Células Madre Neoplásicas/patología , Células U937 , Regulación hacia Arriba , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
Cell adhesion molecule 1 (CADM1/TSLC1) was recently identified as a novel cell surface marker for adult T-cell leukemia/lymphoma (ATLL). In this study, we developed various antibodies as diagnostic tools to identify CADM1-positive ATLL leukemia cells. In flow cytometric analysis, the percentages of CD4(+)CADM1(+) double-positive cells correlated well with both the percentages of CD4(+)CD25(+) cells and with abnormal lymphocytes in the peripheral blood of patients with various types of ATLL. Moreover, the degree of CD4(+)CADM1(+) cells over 1% significantly correlated with the copy number of the human T-lymphotropic virus type 1 (HTLV-1) provirus in the peripheral blood of HTLV-1 carriers and ATLL patients. We also identified a soluble form of CADM1 in the peripheral blood of ATLL patients, and the expression levels of this form were correlated with the levels of soluble interleukin 2 receptor alpha. Moreover, lymphomas derived from ATLL were strongly and specifically stained with a CADM1 antibody. Thus, detection of CD4(+)CADM1(+) cells in the peripheral blood, measurement of serum levels of soluble CADM1 and immunohistochemical detection of CADM1 in lymphomas would be a useful set of markers for disease progression in ATLL and may aid in both the early diagnosis and measurement of treatment efficacy for ATLL.
Asunto(s)
Moléculas de Adhesión Celular/metabolismo , Infecciones por HTLV-I/diagnóstico , Inmunoglobulinas/metabolismo , Leucemia-Linfoma de Células T del Adulto/diagnóstico , Leucemia-Linfoma de Células T del Adulto/metabolismo , Proteínas Supresoras de Tumor/metabolismo , Adulto , Estudios de Casos y Controles , Molécula 1 de Adhesión Celular , Moléculas de Adhesión Celular/inmunología , ADN Viral/genética , Progresión de la Enfermedad , Citometría de Flujo , Infecciones por HTLV-I/genética , Infecciones por HTLV-I/virología , Virus Linfotrópico T Tipo 1 Humano/genética , Humanos , Técnicas para Inmunoenzimas , Inmunoglobulinas/inmunología , Leucemia-Linfoma de Células T del Adulto/virología , Linfocitos/citología , Linfocitos/metabolismo , Provirus/genética , Reacción en Cadena en Tiempo Real de la Polimerasa , Carga ViralRESUMEN
Aberrant activation of Janus kinase 2 (JAK2) caused by somatic mutation of JAK2 (JAK2V617F) or the thrombopoietin receptor (MPLW515L) plays an essential role in the pathogenesis of myeloproliferative neoplasms (MPNs), suggesting that inhibition of aberrant JAK2 activation would have a therapeutic benefit. Our novel JAK2 inhibitor, NS-018, was highly active against JAK2 with a 50% inhibition (IC(50)) of <1 n, and had 30-50-fold greater selectivity for JAK2 over other JAK-family kinases, such as JAK1, JAK3 and tyrosine kinase 2. In addition to JAK2, NS-018 inhibited Src-family kinases. NS-018 showed potent antiproliferative activity against cell lines expressing a constitutively activated JAK2 (the JAK2V617F or MPLW515L mutations or the TEL-JAK2 fusion gene; IC(50)=11-120 n), but showed only minimal cytotoxicity against most other hematopoietic cell lines without a constitutively activated JAK2. Furthermore, NS-018 preferentially suppressed in vitro erythropoietin-independent endogenous colony formation from polycythemia vera patients. NS-018 also markedly reduced splenomegaly and prolonged the survival of mice inoculated with Ba/F3 cells harboring JAK2V617F. In addition, NS-018 significantly reduced leukocytosis, hepatosplenomegaly and extramedullary hematopoiesis, improved nutritional status, and prolonged survival in JAK2V617F transgenic mice. These results suggest that NS-018 will be a promising candidate for the treatment of MPNs.
RESUMEN
Janus kinase 1 (JAK1) and JAK3 plays a critical role in lymphocyte proliferation and differentiation. Somatic JAK1 mutations are found in 18% of adult precursor T acute lymphoblastic leukemias and somatic JAK3 mutations are found in 3.3% of cutaneous T cell lymphomas. Some of the mutations are confirmed as a gain-of-function mutation and are assumed to be involved in leukemogenesis. Adult T cell leukemia/lymphoma (ATLL) is a type of T cell neoplasm, and activation of JAK/STAT pathways is sometimes observed in them. We investigated JAK1 and JAK3 mutations in 20 ATLL patients. No JAK1 mutations were found, and five types of single nucleotide polymorphisms were observed in 12 cases, whose frequencies almost match those in Asian populations. As for JAK3, a synonymous mutation was found in one case. JAK1 and JAK3 mutations are unlikely involved in the leukemogenesis of ATLL.
Asunto(s)
Janus Quinasa 1/genética , Janus Quinasa 3/genética , Leucemia-Linfoma de Células T del Adulto/genética , Adulto , Diferenciación Celular , Proliferación Celular , Análisis Mutacional de ADN , Humanos , Japón , Leucemia-Linfoma de Células T del Adulto/etiología , Polimorfismo de Nucleótido SimpleRESUMEN
The intercalated cell masses of the amygdala consist of cell clusters located between the basolateral complex of the amygdala and its surrounding structures including the central nucleus of the amygdala and the external capsule. Although recent studies have revealed that the intercalated cell masses may play an important role in emotional learning and memory, there are only a few reports on its molecular characterization. We examined the expression patterns of transcription factors in the intercalated cell masses in late embryonic stage and postnatal rats, and non-human primates. Dlx5, Foxp2, Pbx3 and Meis2 were expressed in all subdivisions of the intercalated cell masses, while Ebf1, Nkx2.1 and Foxp1 were not. In contrast, Pax6 was only expressed in a small population of the main intercalated islands, but not in the medial or lateral cell clusters. In addition, few Pax6-positive neurons co-expressed Foxp2. Thus the intercalated cell masses do not contain a homogeneous population of neurons, in terms of their molecular constituents. Given that Foxp2, Pbx3 and Meis2 are preferentially expressed in distinct cell populations in the developing striatum, and that the intercalated cell masses of the amygdala appear to be a ventrocaudal expansion of the striatum, the intercalated neurons may share a common origin with some types of neurons located in the dorsal striatum.
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Amígdala del Cerebelo/embriología , Amígdala del Cerebelo/metabolismo , Cuerpo Estriado/embriología , Cuerpo Estriado/metabolismo , Neuronas/metabolismo , Factores de Transcripción/metabolismo , Amígdala del Cerebelo/citología , Animales , Tipificación del Cuerpo/genética , Cuerpo Estriado/citología , Evolución Molecular , Proteínas del Ojo/genética , Proteínas del Ojo/metabolismo , Femenino , Factores de Transcripción Forkhead/genética , Factores de Transcripción Forkhead/metabolismo , Regulación del Desarrollo de la Expresión Génica/fisiología , Proteínas de Homeodominio/genética , Proteínas de Homeodominio/metabolismo , Macaca , Neurogénesis/fisiología , Neuronas/citología , Factor de Transcripción PAX6 , Factores de Transcripción Paired Box/genética , Factores de Transcripción Paired Box/metabolismo , Proteínas Proto-Oncogénicas/genética , Proteínas Proto-Oncogénicas/metabolismo , Ratas , Ratas Wistar , Proteínas Represoras/genética , Proteínas Represoras/metabolismo , Especificidad de la Especie , Factores de Transcripción/genéticaRESUMEN
BACKGROUND: The F-box protein S-phase kinase-associated protein 2 (Skp2) positively regulates the G1-S transition by promoting degradation of the cyclin-dependent kinase inhibitor p27(kip1) (p27). Recent evidence has indicated an oncogenic role of Skp2 in not only carcinogenesis but also lymphomagenesis. MATERIALS AND METHODS: Clinicopathologic features and immunohistochemical expression of Skp2 and p27 were studied retrospectively in 671 patients treated with cyclophosphamide, vincristine, doxorubicin and prednisolone (CHOP) or cyclophosphamide, vincristine, doxorubicin and prednisolone plus rituximab (R-CHOP). The median follow-up periods were 43.2 months in the CHOP group (n = 425) and 24.0 months in the R-CHOP group (n = 246). RESULTS: High Skp2 or low p27 expression correlated significantly with poor overall survival (OS) and progression-free survival (P < 0.001) in both treatment groups. The prognostic value of Skp2 or p27 expression was independent of the parameters included in the International Prognostic Index by multivariate analysis. Patients with high Skp2 expression in combination with low p27 expression showed the worst survival. CONCLUSIONS: Addition of rituximab to the CHOP regimen did not provide a beneficial outcome to patients with diffuse large B-cell lymphoma with high Skp2 expression and low p27 expression. Skp2 and p27 may be useful prognostic markers in the rituximab era.
Asunto(s)
Anticuerpos Monoclonales/uso terapéutico , Inhibidor p27 de las Quinasas Dependientes de la Ciclina/metabolismo , Linfoma de Células B Grandes Difuso/diagnóstico , Linfoma de Células B Grandes Difuso/tratamiento farmacológico , Proteínas Quinasas Asociadas a Fase-S/metabolismo , Adulto , Anciano , Anciano de 80 o más Años , Anticuerpos Monoclonales/administración & dosificación , Anticuerpos Monoclonales de Origen Murino , Antineoplásicos/administración & dosificación , Antineoplásicos/uso terapéutico , Protocolos de Quimioterapia Combinada Antineoplásica , Biomarcadores de Tumor/metabolismo , Ciclofosfamida , Doxorrubicina , Femenino , Humanos , Linfoma de Células B Grandes Difuso/metabolismo , Linfoma de Células B Grandes Difuso/mortalidad , Masculino , Persona de Mediana Edad , Prednisona , Pronóstico , Estudios Retrospectivos , Rituximab , Análisis de Supervivencia , Vincristina , Adulto JovenRESUMEN
Janus kinase 2 (JAK2) V617F mutation has been regarded as the major cause of myeloproliferative disorders (MPD). However, the mechanisms of abnormal cell growth by JAK2V617F have not been elucidated. In this study, cell cycle regulatory protein expression was analyzed using JAK2V617F-Ba/F3 and mock-Ba/F3. JAK2V617F-Ba/F3, but not mock-Ba/F3, showed IL-3 independent cell growth and constitutive STATs activation. Deregulation of p27(Kip1), the cell cycle regulator at the G1 to S transition, was observed in JAK2V617F-Ba/F3 but not in mock-control. p27(Kip1) deregulation was not due to p27(Kip1) mRNA level but due to high Skp2 expression, a subunit of ubiquitin E3 ligase, through the STAT binding in the Skp2 promoter. Like JAK2V617F overexpression, constitutively active STAT5 or STAT3 induced aberrant p27(Kip1) expression of Ba/F3 cells. Similar findings were observed in BCR/ABL-transfected Ba/F3. Our results elucidate the regulatory mechanism by which JAK2V617F modulates Skp2 gene expression through the STAT transcription factors.
Asunto(s)
Inhibidor p27 de las Quinasas Dependientes de la Ciclina/metabolismo , Regulación de la Expresión Génica , Janus Quinasa 2/metabolismo , Proteínas Quinasas Asociadas a Fase-S/genética , Factores de Transcripción STAT/metabolismo , Animales , Línea Celular Tumoral , Ensayo de Cambio de Movilidad Electroforética , Interleucina-3/metabolismo , Janus Quinasa 2/genética , Ratones , Mutación , Regiones Promotoras GenéticasRESUMEN
beta-Thujaplicin (hinokitiol) is a tropolone derivative present in the heartwood of cupressaceous plants and is used as a medicine, a food additive, and a preservative, and in cosmetics as hair tonic. The cultured plant cells of Nicotiana tabacum glycosylated beta-thujaplicin to two glucosides, 4-isopropyltropolone 2-O-beta-D-glucoside (6%) and 6-isopropyltropolone 2-O-beta-D-glucoside (12%), and two gentiobiosides, 4-isopropyltropolone 2-O-beta-D-gentiobioside (2%) and 6-isopropyltropolone 2-O-beta-D-gentiobioside (5%) after 48 h incubation. The use of immobilized cells of N. tabacum in sodium alginate gel much improved the yield of the products; the glycosylation of beta-thujaplicin with immobilized N. tabacum gave the glycoside products, 4-isopropyltropolone 2-O-beta-D-glucoside (11%), 4-isopropyltropolone 2-O-beta-D-gentiobioside (6%), 6-isopropyltropolone 2-O-beta-D-glucoside (20%), and 6-isopropyltropolone 2-O-beta-D-gentiobioside (10%). On the other hand, 4-isopropyltropolone 2-O-beta-D-glucoside (14%), 4-isopropyltropolone 2-O-beta-D-gentiobioside (7%), 6-isopropyltropolone 2-O-beta-D-glucoside (33%), and 6-isopropyltropolone 2-O-beta-D-gentiobioside (13%) were obtained through the biotransformation with immobilized cells in the medium without iron ions. In comparison with the case of bioconversion in the normal medium containing iron ions, removal of iron ions improved the yields of products.
Asunto(s)
Glicósidos/metabolismo , Monoterpenos/metabolismo , Nicotiana/metabolismo , Tropolona/análogos & derivados , Línea Celular , Estructura Molecular , Monoterpenos/química , Tropolona/química , Tropolona/metabolismoRESUMEN
BACKGROUND: Psychiatric comorbidity of autism spectrum disorder (ASD) has not been well examined. METHODS: Mood disorders in 44 consecutive outpatients with high-functioning ASD were examined at a university hospital according to DSM-IV. Inclusion criteria were an IQ of 70 or higher on the Wechsler Intelligence Scale and age of 12 years or over. RESULTS: Sixteen patients (36.4%) were diagnosed with mood disorder. Of these 16 patients, four were diagnosed as having major depressive disorder, two patients as bipolar I disorder, six patients as bipolar II disorder, and four patients as bipolar disorder not otherwise specified. Bipolar disorder accounted for 75% of cases. Twelve patients had Asperger disorder and four patients had pervasive developmental disorder not otherwise specified. None of the patients had autistic disorder. LIMITATIONS: The sample size was small. We could not use Autism Diagnostic Interview - Revised. Referral bias could not be avoided in this study. CONCLUSIONS: The major comorbid mood disorder in patients with high-functioning ASD is bipolar disorder and not major depressive disorder. The autistic spectrum may share common vulnerability genes with the bipolar spectrum.
Asunto(s)
Atención Ambulatoria , Síndrome de Asperger/epidemiología , Trastorno Autístico/epidemiología , Trastorno Bipolar/epidemiología , Trastornos Generalizados del Desarrollo Infantil/epidemiología , Adolescente , Adulto , Síndrome de Asperger/diagnóstico , Trastorno Autístico/diagnóstico , Trastorno Bipolar/diagnóstico , Niño , Trastornos Generalizados del Desarrollo Infantil/diagnóstico , Comorbilidad , Trastorno Depresivo Mayor/diagnóstico , Trastorno Depresivo Mayor/epidemiología , Manual Diagnóstico y Estadístico de los Trastornos Mentales , Femenino , Estudios de Seguimiento , Humanos , Masculino , Prevalencia , Escalas de Valoración Psiquiátrica , Encuestas y Cuestionarios , Escalas de Wechsler , Adulto JovenRESUMEN
An acquired JAK2 V617F mutation is found in most patients with polycythemia vera (PV), and about half of patients with essential thrombocythemia (ET) or primary myelofibrosis (PMF). Mice transplanted with bone marrow cells in which JAK2 V617F was retrovirally expressed developed PV-like features, but not ET or PMF. To address the contribution of this mutation to the pathogenesis of these three MPDs, we generated two lines of JAK2 V617F transgenic mice. One line showed granulocytosis after 4 months of age. Among 43 mice, 8 (19%) showed polycythemia and 15 (35%) showed thrombocythemia. The second line showed extreme leukocytosis and thromobocytosis. They showed anemia that means Hb value from 9 to 10 g per 100 ml when 1 month old. Myeloid cells and megakaryocytes were predominant in the bone marrow of these animals, and splenomegaly was observed. The expression of JAK2 V617F mRNA in bone marrow cells was 0.45 and 1.35 that of endogenous wild-type JAK2 in the two lines, respectively. In vitro analysis of bone marrow cells from both lines showed constitutive activation of ERK1/2, STAT5 and AKT, and augmentation of their phosphorylations by cytokine stimulation. We conclude that in vivo expression of JAK2 V617F results in ET-, PMF- and PV-like disease.