RESUMEN
Somatic PTPN11 mutations cause juvenile myelomonocytic leukemia (JMML). Germline PTPN11 defects cause Noonan syndrome (NS), and specific inherited mutations cause NS/JMML. Here, we report that hematopoietic cells differentiated from human induced pluripotent stem cells (hiPSCs) harboring NS/JMML-causing PTPN11 mutations recapitulated JMML features. hiPSC-derived NS/JMML myeloid cells exhibited increased signaling through STAT5 and upregulation of miR-223 and miR-15a. Similarly, miR-223 and miR-15a were upregulated in 11/19 JMML bone marrow mononuclear cells harboring PTPN11 mutations, but not those without PTPN11 defects. Reducing miR-223's function in NS/JMML hiPSCs normalized myelogenesis. MicroRNA target gene expression levels were reduced in hiPSC-derived myeloid cells as well as in JMML cells with PTPN11 mutations. Thus, studying an inherited human cancer syndrome with hiPSCs illuminated early oncogenesis prior to the accumulation of secondary genomic alterations, enabling us to discover microRNA dysregulation, establishing a genotype-phenotype association for JMML and providing therapeutic targets.
Asunto(s)
Células Madre Pluripotentes Inducidas/citología , Leucemia Mielomonocítica Juvenil/metabolismo , Células Mieloides/citología , Proteína Tirosina Fosfatasa no Receptora Tipo 11/genética , Lectina 3 Similar a Ig de Unión al Ácido Siálico/metabolismo , Células Cultivadas , Células HEK293 , Humanos , Células Madre Pluripotentes Inducidas/metabolismo , Leucemia Mielomonocítica Juvenil/genética , Leucemia Mielomonocítica Juvenil/patología , MicroARNs/genética , Mutación , Células Mieloides/metabolismo , Lectina 3 Similar a Ig de Unión al Ácido Siálico/genética , Regulación hacia ArribaRESUMEN
The conclusions reached by a diverse group of scientists who attended an intense 2-day workshop on hybrid organic-inorganic perovskites are presented, including their thoughts on the most burning fundamental and practical questions regarding this unique class of materials, and their suggestions on various approaches to resolve these issues.
RESUMEN
The interpretation of Onsager cross transport coefficients measured in mixed ionic electronic conductor (MIEC) oxides is examined. It is demonstrated that the cross terms are an artifact of the way the measurements are analyzed. When an appropriate, comprehensive defect model is considered for the MIEC, no cross terms are required.
RESUMEN
Dilated cardiomyopathy (DCM) is a highly heterogeneous trait with sarcomeric gene mutations predominating. The cause of a substantial percentage of DCMs remains unknown, and no gene-specific therapy is available. On the basis of resequencing of 513 DCM cases and 1,150 matched controls from various cohorts of distinct ancestry, we discovered rare, functional RAF1 mutations in 3 of the cohorts (South Indian, North Indian and Japanese). The prevalence of RAF1 mutations was ~9% in childhood-onset DCM cases in these three cohorts. Biochemical studies showed that DCM-associated RAF1 mutants had altered kinase activity, resulting in largely unaltered ERK activation but in AKT that was hyperactivated in a BRAF-dependent manner. Constitutive expression of these mutants in zebrafish embryos resulted in a heart failure phenotype with AKT hyperactivation that was rescued by treatment with rapamycin. These findings provide new mechanistic insights and potential therapeutic targets for RAF1-associated DCM and further expand the clinical spectrum of RAF1-related human disorders.
Asunto(s)
Cardiomiopatía Dilatada/genética , Mutación , Proteínas Proto-Oncogénicas c-raf/genética , Adulto , Edad de Inicio , Anciano , Secuencia de Aminoácidos , Animales , Cardiomiopatía Dilatada/etnología , Estudios de Casos y Controles , Estudios de Cohortes , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Femenino , Fibroblastos/metabolismo , Células HEK293 , Humanos , India , Japón , Masculino , Ratones , Persona de Mediana Edad , Datos de Secuencia Molecular , Fenotipo , Prevalencia , Homología de Secuencia de Aminoácido , Sirolimus/química , Pez CebraRESUMEN
In order to study the effects of Hepatocyte Growth Factor (HGF) in the heart, two transgenic mice were developed, one carrying a bidirectional HGF-TetO-GFP responder construct and the other carrying a α-MHC-tTA transactivator construct. Crosses were carried out between heterozygotes, so that litters contained bitransgenic α-MHC-tTA/HGF-TetO-GFP+, thus expressing HGF and GFP exclusively in the heart and only in the absence of Doxycycline. Our data show that the expression of HGF was indeed restricted to the heart and that the expression was limited to the timeframe of the absence of Doxycycline. Surprisingly the expression was variable even between bitransgenic littermates. In the setting of a model of ischemia-reperfusion, the expression of HGF ameliorates cardiac functionality, enhances proliferation and diminishes the scarred area, proving that this is a good model to study the beneficial influences and functional roles of HGF in the heart.
Asunto(s)
Doxiciclina/farmacología , Corazón/fisiopatología , Factor de Crecimiento de Hepatocito/metabolismo , Animales , Western Blotting , Línea Celular , Proliferación Celular , Colágeno/metabolismo , Cruzamientos Genéticos , Medios de Cultivo Condicionados/metabolismo , Perros , Ecocardiografía , Femenino , Regulación de la Expresión Génica , Proteínas Fluorescentes Verdes/metabolismo , Corazón/efectos de los fármacos , Factor de Crecimiento de Hepatocito/genética , Heterocigoto , Inmunohistoquímica , Ratones , Ratones Transgénicos , Modelos Animales , Daño por Reperfusión Miocárdica/tratamiento farmacológico , Daño por Reperfusión Miocárdica/fisiopatología , Embarazo , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
BACKGROUND: The Hepatocyte Growth Factor (HGF) is a pleiotropic cytokine involved in many physiological processes, including skeletal muscle, placenta and liver development. Little is known about its role and that of Met tyrosine kinase receptor in cardiac development. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we generated two transgenic mice with cardiac-specific, tetracycline-suppressible expression of either Hepatocyte Growth Factor (HGF) or the constitutively activated Tpr-Met kinase to explore: i) the effect of stimulation of the endogenous Met receptor by autocrine production of HGF and ii) the consequence of sustained activation of Met signalling in the heart. We first showed that Met is present in the neonatal cardiomyocytes and is responsive to exogenous HGF. Exogenous HGF starting from prenatal stage enhanced cardiac proliferation and reduced sarcomeric proteins and Connexin43 (Cx43) in newborn mice. As adults, these transgenics developed systolic contractile dysfunction. Conversely, prenatal Tpr-Met expression was lethal after birth. Inducing Tpr-Met expression during postnatal life caused early-onset heart failure, characterized by decreased Cx43, upregulation of fetal genes and hypertrophy. CONCLUSIONS/SIGNIFICANCE: Taken together, our data show that excessive activation of the HGF/Met system in development may result in cardiac damage and suggest that Met signalling may be implicated in the pathogenesis of cardiac disease.
Asunto(s)
Cardiopatías/metabolismo , Corazón/crecimiento & desarrollo , Miocardio/enzimología , Miocardio/patología , Proteínas Proto-Oncogénicas c-met/metabolismo , Transducción de Señal , Animales , Animales Recién Nacidos , Proliferación Celular/efectos de los fármacos , Conexina 43/metabolismo , Activación Enzimática/efectos de los fármacos , Regulación de la Expresión Génica/efectos de los fármacos , Corazón/fisiopatología , Cardiopatías/enzimología , Cardiopatías/etiología , Cardiopatías/patología , Factor de Crecimiento de Hepatocito/genética , Factor de Crecimiento de Hepatocito/farmacología , Ratones , Ratones Transgénicos , Contracción Muscular/efectos de los fármacos , Proteínas Musculares/metabolismo , Miocardio/metabolismo , Miocitos Cardíacos/efectos de los fármacos , Miocitos Cardíacos/metabolismo , Miocitos Cardíacos/patología , Especificidad de Órganos , Transducción de Señal/efectos de los fármacosRESUMEN
We show that within the validity range of local thermal equilibrium--therein, however, irrespective of the magnitude of the driving force--a simple current equation can be formulated that expresses the current in terms of a product of a local nonequilibrium conductivity and a sinh function of half the electrochemical potential drop (normalized with respect to kBT) over the local hopping distance. This local current/driving force relation takes account of both electrical and compositional effects and can be generalized as to include interactions and structural variations.
Asunto(s)
Modelos Teóricos , Electroquímica , Campos Electromagnéticos , Cinética , Sustancias MacromolecularesRESUMEN
PAX3-FKHR, the product of a rearrangement of PAX3 with FKHR is the pathogenetic marker for alveolar rhabdomyosarcoma, an aggressive form of childhood cancer. In this work we show that PAX3-FKHR, which is a stronger transcriptional activator relative to PAX3, can lead to two apparently irreconcilable outcomes: transformation and terminal myogenic differentiation. Fibroblasts (10T1/2, NIH3T3, and a newly established murine line named 'Plus') transduced by PAX3-FKHR acquire transformed features such as anchorage independence and loss of contact inhibition and concomitantly undergo various degrees of myogenic conversion depending on the host cells, including, in the case of the Plus line, terminal differentiation into contractile myotubes. This work highlights the potential of PAX3-FKHR to functionally operate as a deregulated Pangene and may have implications with regard to the identity of the precursor cell giving rise to alveolar rhabdomyosarcoma.