ABSTRACT
TAF15 (formerly TAFII68) is a member of the FET (FUS, EWS, TAF15) family of RNA- and DNA-binding proteins whose genes are frequently translocated in sarcomas. By performing global gene expression profiling, we found that TAF15 knockdown affects the expression of a large subset of genes, of which a significant percentage is involved in cell cycle and cell death. In agreement, TAF15 depletion had a growth-inhibitory effect and resulted in increased apoptosis. Among the TAF15-regulated genes, targets of microRNAs (miRNAs) generated from the onco-miR-17 locus were overrepresented, with CDKN1A/p21 being the top miRNAs-targeted gene. Interestingly, the levels of onco-miR-17 locus coded miRNAs (miR-17-5p and miR-20a) were decreased upon TAF15 depletion and shown to affect the post-transcriptional regulation of TAF15-dependent genes, such as CDKN1A/p21. Thus, our results demonstrate that TAF15 is required to regulate gene expression of cell cycle regulatory genes post-transcriptionally through a pathway involving miRNAs. The findings that high TAF15 levels are needed for rapid cellular proliferation and that endogenous TAF15 levels decrease during differentiation strongly suggest that TAF15 is a key regulator of maintaining a highly proliferative rate of cellular homeostasis.
Subject(s)
Cell Cycle/physiology , Cell Division/physiology , Gene Expression Regulation , TATA-Binding Protein Associated Factors/physiology , Apoptosis/physiology , Cell Differentiation , Cell Line, Tumor/drug effects , Cyclin-Dependent Kinase Inhibitor p21/biosynthesis , Cyclin-Dependent Kinase Inhibitor p21/genetics , Gene Expression Profiling , Gene Regulatory Networks , Genes, Reporter , HeLa Cells , Humans , MicroRNAs/biosynthesis , MicroRNAs/genetics , Neoplasm Proteins/antagonists & inhibitors , Neoplasm Proteins/biosynthesis , Neoplasm Proteins/genetics , Neoplasm Proteins/physiology , Neuroblastoma/pathology , Neurogenesis , Neurons/cytology , RNA Interference , TATA-Binding Protein Associated Factors/antagonists & inhibitors , TATA-Binding Protein Associated Factors/geneticsABSTRACT
Los síndromes poliendocrinos autoinmunes (APS) asocian enfermedades endocrinas autoinmunes con otros desórdenes autoinmunes no endocrinos. El APS tipo II se caracteriza por compromiso primario suprarrenal, tiroideo y/o DM tipo I. Presentamos un paciente masculino de 46 años que fue internado por astenia, adinamia, hiporexia, severa disminución de peso, mareos y vómitos. Antecedente de obesidad y diabetes diagnosticada 3 años antes. Presentaba hipotensión arterial, hiperpigmentación de mucosas y pliegues, anemia, hiponatremia e hipoglucemias frecuentes a pesar de la disminución de la dosis de insulina. Se diagnosticó insuficiencia suprarrenal, concomitantemente con hipotiroidismo y diabetes tipo 1, todas de origen autoinmune, iniciándose reemplazo hormonal. Se encontró una posible asociación del HLA DRB1*-DQB1* en los estudios genéticos. Conclusiones: Nuestro paciente presenta el HLA DQB1*0302 descripto en el APSII, pero el HLA DRB1 *08 encontrado no está descripto en este síndrome ni en ningún otro desorden autoinmune. En pacientes con Diabetes tipo 1 que disminuyan el requerimiento insulínico, habría que descartar insuficiencia suprarrenal, un componente del APS II, como factor etiológico, a pesar de su baja prevalencia.
Autoimmune polyendocrine syndromes (APS) are the association of autoimmune endocrine diseases with other non-endocrine autoimmune disorders. Type II APS is defined by occurrence of Addison´s disease with thyroid autoimmune disease and/or type 1 diabetes mellitus. We present a 46-year-old male patient who was hospitalized because of asthenia, adynamia, hyporexia, severe loss of weight, dizziness and vomiting. Diabetes mellitus had been diagnosed 3 years earlier when he was obese. He presented arterial hypotension, anemia, darkening of the skin and oral mucosa, hyponatremia and frequent hypoglycemia although his insulin dose was decreased. Adrenal insufficiency was diagnosed together with hypothyroidism and type 1 diabetes, all of them of autoimmune origin. Hormonal replacement treatment was initiated. Genetic studies were performed and a new polymorphism was found. Conclusions: HLA DRB1 *08 found in our patient has not been described in APS II or in any other autoimmune disorders. He also has HLA DQB1*0302 described in previous reports related to APS II. In type 1 diabetic patients whose insulin requirement decreases, it would be advisable to rule out adrenal insufficiency, a component of APS II, as an etiologic factor in spite of its low prevalence. In diabetic obese patients (mainly young) who lose weight without a defined cause, type 1 diabetes mellitus should be excluded.
Subject(s)
Humans , Male , Middle Aged , Polyendocrinopathies, Autoimmune/complications , Polyendocrinopathies, Autoimmune/diagnosis , Polyendocrinopathies, Autoimmune/immunology , Genetic Association StudiesABSTRACT
Introducción: La determinación de IGF-I en suero o plasma es una herramienta esencial en el diagnóstico y seguimiento de la acromegalia. Sin embargo, se deben tener presentes algunos inconvenientes en su medición por diferentes inmunoensayos. Objetivos: Evaluar dos inmunoensayos para la determinación de IGF-I y su correlación con el nadir de GH en el TTOG en pacientes acromegalicos. Materiales y métodos: Se analizaron 37 pacientes acromegálicos, 20 mujeres y 17 hombres. IGF-I fue determinada por Immulite 1000, (IMM) y por IRMA (DSL). Se realizó el TTOG y se determinó glucosa y GH en todos los tiempos (basal, 30, 60, 90 y 120min). Se consideró respuesta normal un nadir de GH <1ng/ml. Nueve pacientes se encontraban bajo tratamiento y 28 sin tratamiento. Análisis estadístico: se utilizaron el test de Wilcoxon, de Bland y Altman y curvas ROC. Se consideró significativa una p<0,05. Resultados: Las concentraciones basales de glucosa fueron 97,86±10,91 mg/dl, de GH 2,8 (1,59-14,4) ng/ml, de IGF-I por IMM 602±318 ng/ml y por DSL 1006±596 ng/ml. IGF-I por IMM y DSL mostró una diferencia significativa con p <0,01 y un bias de - 403,2 ng/ml con valores menores por IMM. IGF-I elevada por IMM y DSL, se encontró en el 84% y en el 97% respectivamente. IGF-I elevada con nadir de GH >1ng/ml se encontró en el 70%, con nadir de GH normal en el 13,5%. IGF-I normal con nadir >1ng/ml en el 2,7% y con nadir de GH normal en el 13,5%. El área bajo las curvas ROC no mostró diferencias significativas. Conclusiones: Los niveles de IGF-I determinados por IMM y DSL fueron significativamente diferentes mostrando un bias negativo para IMM. La mayoría de los valores del nadir de GH fueron consistentes con los niveles de IGF-I observándose una discrepancia en el 30% de los pacientes, estuvieran o no bajo tratamiento.
Introduction: IGF-I determination in serum or plasma is an essential tool in the diagnosis and follow-up of acromegaly. Hepatic production of IGF-I is regulated by GH and circulates bound to several IGF-I binding proteins which extends its half life. IGF-I is not released in a pulsatile pattern and has no significant variability in 24 h. Objective: To evaluate two different methodologies in IGF-I levels determination and their correlation with GH nadir in OGTT in acromegalic patients. Material and methods: We analyzed 37 acromegalic patients, 20 women and 17 men, mean age was 45±12 years. IGF-I levels were assayed by Immulite 1000, DPC (IMM) and DSL-5600 ACTIVE® IGF-I Coated-Tube IRMA (DSL) and OGTTs (at baseline and at 30, 60, 90 and 120 minutes) were performed by measuring plasma glucose and GH assay by immunochemiluminometric assay (Access); we considered a nadir <1ng/ml as normal response. Nine patients were under medical treatment (cabergoline: 4, octeotride: 4, and cabergoline plus octeotrite: 1) and 28 without treatment. Statistical analysis: Wilcoxon and, Bland and Altman tests and ROC curves. Differences were considered significant at p< 0.05. Results: Basal glucose levels were 97.86±10.91 mg/dl and mean GH was 2.8 (1.59-14.4) ng/ml. Mean IGF-I levels performed by IMM were 602±318 ng/ml and 1006±596 ng/ml by DSL. There was a statistically significant difference between both methodologies (p<0.01). Bland and Altman test showed a bias of - 403.2 ng/ml with lower values by IMM. We observed elevated IGF-I levels in 84% by IMM and in 97% by DSL, and only one patient had normal levels with both methodologies. Elevated IGF-I levels and GH nadir >1ng/ml were observed in 70% of the patients, increased IGF-I with normal GH nadir in 13.5%, normal IGF-I with GH nadir >1ng/ml in 2.7% and normal IGF-I with normal GH nadir in 13.5%. Patients under treatment: 3 showed normal GH nadir with elevated IGF-I levels, in 2 of them by both methodologies, and in the other one it was normal by IMM and elevated by DSL; the other 6 showed GH nadir > 1ng/ml, 5 of them presented elevated IGF-I by both methodologies and the other one showed discrepancy in IGF-I levels. The under ROC curve area and confidence interval (CI) of 95% for IGF-I IMM and DSL were 0.96 (0.90-1.00) and 0.91 (0.82-1.00) respectively. Differences between the ROC curves areas were not significant Conclusions: IGF-I levels determined by IMM and DSL were statistically significantly different. IGF-I levels showed a negative bias by IMM. Most of the results of GH nadir were consistent with IGF-I levels but we observed discrepancy in 30% of the patients, regardless of whether they were under treatment or not.
Subject(s)
Humans , Male , Female , Child , Adolescent , Adult , Middle Aged , Acromegaly/blood , Insulin-Like Growth Factor I/analysis , Glucose Tolerance Test/statistics & numerical data , Immunoassay/methods , Data Interpretation, Statistical , Human Growth Hormone/analysisABSTRACT
Objetivos: Estimar la frecuencia de complicaciones maternofetales en mujeres que se embarazaron durante el tratamiento con cabergolina (CAB). Estimar la frecuencia de patología detectada posnacimiento en los niños producto de dichos embarazos. Material y métodos: Estudio retrospectivo y multicéntrico de 86 embarazos en 78 mujeres con hiperprolactinemia idiopática (7) o tumoral (44 micro y 27 macro), en tratamiento con CAB en el momento de la concepción. Edad: 20 a 45 años; PRL inicial: 30 a 1429 ng/ml; duración del tratamiento previo al embarazo 1 a 120 meses; dosis: 0.125 a 4 mg/semana. El rango de exposición embriofetal a la CAB fue de 3 a 27 semanas, el 96.39% de las pacientes la recibió durante el primer trimestre y el 3.61% hasta el segundo. Resultados: No hubo complicaciones mayores durante el embarazo. Se registraron 7 abortos espontáneos (8.1%) y 75 partos, de los cuales 49 fueron vaginales y 26 cesáreas. Se registraron 69 recién nacidos, 63 fueron a término y 6 pretérmino (8.8%), ninguno bajo peso para la edad gestacional. En 3 (5.2%) recién nacidos se observó: 1 malformación mayor (Síndrome de Down) y 2 menores (hernia umbilical e inguinal). Se obtuvo seguimiento de 42 recién nacidos; se diagnosticó epilepsia refractaria en uno y un trastorno generalizado del desarrollo en otro. No se halló una mayor frecuencia de complicaciones en los embarazos ni en los recién nacidos expuestos a CAB que en la población normal. Sería necesario mayor número de pacientes para concluir sobre la seguridad de CAB durante el embarazo.
Objectives: To assess the rate of any potential adverse effects on pregnancy and embryo-fetal development in women who became pregnant under treatment with cabergoline (CAB). To follow up medical data of children who were born from mothers exposed to Cab in early weeks of gestation. Material and methods: Observational, retrospective and multicenter study on 86 pregnancies in 78 women with idiopathic or tumoral hyperprolactinemia. All patients were under Cab at conception. The average age was 29 (range: 20-45). Pituitary images at diagnosis showed 44 microadenomas, 27 macroadenomas and 7 were normal. Serum PRL at baseline was between 30 and 1429 ng/ml. Duration of therapy before pregnancy ranged from 1 to 120 months. Maternal and fetal exposure to cabergoline and doses ranged from 0.125 to 4 mg/week. The mean serum PRL level under which patients achieved pregnancy was 17 ng/ml. Fetal exposure ranged from 3 to 27 weeks; 96.39% of patients received CAB during the first trimester of pregnancy and 3.61% until the second one. Results: No significant complications during pregnancy were found. Seven women (8.1%) had spontaneous abortions. Term deliveries were recorded in 63/69, preterm in six (8.8%), none of them with low weight for gestational age. Neonatal abnormalities were observed in 3 (5.2%): 1 major (Down syndrome) and 2 minor malformations (umbilical and inguinal hernia). Two out of 42, developed abnormalities during the follow- up, one of them was a refractory epilepsy during the second month of life, the other presented a Pervasive Developmental Disorder diagnosed in the third year of life. Conclusion: No significantly higher frequency of complications was found in pregnancies and/or offspring exposed to CAB than in normal population. Larger series of patients are needed to asses the safety.
Subject(s)
Humans , Female , Pregnancy , Adult , Middle Aged , Pregnancy Complications/etiology , Ergolines/adverse effects , Congenital Abnormalities/prevention & control , Pregnancy/drug effects , Embryonic and Fetal Development/drug effectsABSTRACT
In the acute promyelocytic leukemia (APL) bearing the t(15;17), all-trans-retinoic acid (ATRA) treatment induces granulocytic maturation and complete remission of leukemia. We identified miR-342 as one of the microRNAs (miRNAs) upregulated by ATRA during APL differentiation. This miRNA emerged as a direct transcriptional target of the critical hematopoietic transcription factors PU.1 and interferon regulatory factor (IRF)-1 and IRF-9. IRF-1 maintains miR-342 at low levels, whereas the binding of PU.1 and IRF-9 in the promoter region following retinoic ATRA-mediated differentiation, upregulates miR-342 expression. Moreover, we showed that enforced expression of miR-342 in APL cells stimulated ATRA-induced differentiation. These data identified miR-342 as a new player in the granulocytic differentiation program activated by ATRA in APL.
Subject(s)
Cell Differentiation , Granulocytes/cytology , Interferon Regulatory Factor-1/genetics , Interferon-Stimulated Gene Factor 3, gamma Subunit/genetics , Leukemia, Promyelocytic, Acute/genetics , MicroRNAs/genetics , MicroRNAs/physiology , Proto-Oncogene Proteins/genetics , Trans-Activators/genetics , Tretinoin/pharmacology , Antineoplastic Agents/pharmacology , Cell Adhesion Molecules/genetics , Cell Adhesion Molecules/metabolism , Chromatin Immunoprecipitation , Granulocytes/drug effects , Granulocytes/metabolism , Humans , Immunoblotting , Immunophenotyping , Interferon Regulatory Factor-1/metabolism , Interferon-Stimulated Gene Factor 3, gamma Subunit/metabolism , Introns/genetics , Leukemia, Promyelocytic, Acute/metabolism , Leukemia, Promyelocytic, Acute/pathology , Promoter Regions, Genetic , Proto-Oncogene Proteins/metabolism , RNA, Messenger/genetics , RNA, Messenger/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Trans-Activators/metabolism , Tumor Cells, CulturedABSTRACT
We describe a pathway by which the master transcription factor PU.1 regulates human monocyte/macrophage differentiation. This includes miR-424 and the transcriptional factor NFI-A. We show that PU.1 and these two components are interlinked in a finely tuned temporal and regulatory circuitry: PU.1 activates the transcription of miR-424, and this up-regulation is involved in stimulating monocyte differentiation through miR-424-dependent translational repression of NFI-A. In turn, the decrease in NFI-A levels is important for the activation of differentiation-specific genes such as M-CSFr. In line with these data, both RNAi against NFI-A and ectopic expression of miR-424 in precursor cells enhance monocytic differentiation, whereas the ectopic expression of NFI-A has an opposite effect. The interplay among these three components was demonstrated in myeloid cell lines as well as in human CD34+ differentiation. These data point to the important role of miR-424 and NFI-A in controlling the monocyte/macrophage differentiation program.
Subject(s)
Cell Differentiation , Hematopoiesis , Macrophages/cytology , Macrophages/metabolism , MicroRNAs/genetics , Monocytes/cytology , Monocytes/metabolism , Proto-Oncogene Proteins/metabolism , Trans-Activators/metabolism , Base Sequence , Cells, Cultured , Humans , NFI Transcription Factors/genetics , NFI Transcription Factors/metabolism , Protein Binding , Up-RegulationABSTRACT
The discovery of microRNAS (miRNAs) and of their mechanism of action has provided some very new clues on how gene expression is regulated. These studies established new concepts on how posttranscriptional control can fine-tune gene expression during differentiation and allowed the identification of new regulatory circuitries as well as factors involved therein. Because of the wealth of information available about the transcriptional and cellular networks involved in hematopoietic differentiation, the hematopoietic system is ideal for studying cell lineage specification. An interesting interplay between miRNAs and lineage-specific transcriptional factors has been found, and this can help us to understand how terminal differentiation is accomplished.
