IL-5 blocks apoptosis and tau hyperphosphorylation induced by Abeta25-35 peptide in PC12 cells

The primary features of Alzheimer’s disease (AD) are extracellular amyloid plaques consisting mainly of deposits of amyloid β (Aβ) peptides and intracellular neurofibrillary tangles (NFTs). Sets of evidence suggest that interleukin-5 (IL-5) is involved in the pathogenesis of AD. Herein, we investigated the protective role of IL-5 in PC12 cells, to provide new insights into understanding this disease. Western blot was employed to assess the protein levels of Bax and phospho-tau as well as phospho-JAK2; MTT assay was performed to decipher cell viability. Treatment of IL-5 decreased Aβ25-35-induced tau phosphorylation and apoptosis, effects blunted by JAK2 inhibition. IL-5 prevents Aβ25-35-evoked tau protein hyperphosphorylation and apoptosis through JAK2 signaling (AU)

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MicroRNA-9 regulates cardiac fibrosis by targeting PDGFR-β in rats

The proliferation of cardiac fibroblasts (CFs) and excessive deposition of extracellular matrix (ECM) are the main pathological characteristics of cardiac fibrosis. In recent years, microRNAs (miRNAs) have been found to be a new kind of regulator in cardiac fibrosis. The purpose of this study was to investigate the role of microRNA-9 (miR-9) in the process of cardiac fibrosis and its mechanism. Treatment of cultured neonatal rat CFs with PDGF-BB or serum suppressed the expression of miR-9. Overexpression of miR-9 obviously inhibited neonatal rat CFs proliferation and collagen production as detected by MTT assays, qRT-PCR, and western blotting. The effects of miR-9 in CFs were abrogated by co-transfection with miR-9 inhibitors. Overexpression of miR-9 reduced the mRNA and protein levels of PDGFR-βand its downstream protein, extracellular signal-regulated kinase (ERK) 1/2. Silencing PDGFR-βby small interfering RNA mimicked the anti-fibrotic action of miR-9, whereas overexpression of PGDFR-β canceled the effect of miR-9 in cultured CFs. Dual-luciferase reporter assays showed that PDGFR-βwas a direct target of miR-9. Overexpression of miR-9 inhibited cardiac fibrosis by targeting PDGFR-β, indicating that miR-9 might play a role in the treatment of cardiac fibrosis (AU)

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Overexpression of forkhead box protein M1 (FOXM1) plays a critical role in colorectal cancer

Background: The forkhead box M1 (FOXM1), an important regulator of cell differentiation and proliferation, is overexpressed in a number of aggressive human carcinomas. However, the clinical significance of FOXM1 signaling in human colorectal cancer (CRC) pathogenesis remains unknown. The aim of this study was to evaluate the role of FOXM1 in CRC tumorigenesis. Methods: We investigated FOXM1 expression in 103 cases of primary CRC and matched normal tissue specimens and explored the underlying mechanisms of altered FOXM1 expression and the impact of this altered expression on CRC proliferation and metastasis using in vitro models of CRC. Results: The results showed that high expression of FOXM1 staining was 85.44 % (88/103) in 103 cases of CRC and 20.39 % (21/103) in 103 cases of adjacent noncancerous tissue samples; the difference of FOXM1 expression between two groups was statistically significant (P < 0.001). Silencing of FOXM1 inhibited the proliferation of CRC cells, and the invasion and migration of CRC cells were distinctly suppressed. Furthermore, FOXM1 knockdown led to substantial reductions in VEGF-A levels in CRC cell lines. Conclusions: Our data suggest that the pathogenesis of CRC maybe mediated by FOXM1, and FOXM1 could represent selective targets for the molecularly targeted treatments of CRC

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Sistemas genéticos para un nuevo abordaje del riesgo de progresión de la retinopatía diabética / Genetic systems for a new approach to risk of progression of diabetic retinopathy

OBJETIVO: Evaluar el riesgo de progresión de la retinopatía diabética (RD) utilizando nuevas estrategias para obtener información genética en diabéticos tipo 2 (DT2) basadas en interferencia por ácido ribonucleico (ARN). MATERIAL Y MÉTODOS: Estudio multicéntrico, prospectivo de casos-controles en 132 participantes divididos en: grupo DT2 (GDT2) con RD (+RD) y sin RD (-RD) (n = 77) y grupo control (GC) (n = 55). Tras entrevista personal y examen oftalmológico, se extrajeron lágrimas para análisis molecular (expresión de micro-ARN [miARN] [miRCURY™ ARN Isolation Kit, Qiagen]). En 18 muestras (GDT2+RD = 6; GDT2-RD = 6; GC = 6) obtuvimos librerías de 137 vs. 140 pares de bases (GeneMapper, Applied Biosystems) y realizamos secuenciación de próxima generación (NGS). El programa SPSS 15.0 vehiculizó el análisis estadístico. RESULTADOS: Edad media: 67 ± 12 años en GDT2 vs. 55 ± 21 años en GC. Distribución hombres/mujeres: 51/28 en GDT2 vs. 25/30 en GC. Los antecedentes familiares de DM, cumplir dieta, fumar, beber y realizar ejercicio mostraron diferencias significativas entre grupos (p < 0,001). Con 20-25 μL de lágrimas extrajimos 9,42 ± 3,30 ng/mL de ARN purificado, con diferencias significativas entre GDT2/GC (p = 0,002) y GDT2+RD/GC (p = 0,004). La expresión lagrimal de miARN en GDT2 correlacionó directamente con: edad/obesidad/duración de DM (p < 0,05), e indirectamente con: agudeza visual (p < 0,05). Hemos identificado 14 miARN relacionados con la presencia, mecanismos patogénicos y factores de riesgo para la progresión de la RD. CONCLUSIONES: Proponemos utilizar lágrimas como fuente de información genética para la DM. Los miARN específicos implicados en desarrollo o progresión de la RD pueden utilizarse como biomarcadores moleculares y, a partir de ellos, desarrollar futuras bioterapias

OBJECTIVE: To evaluate the risk of progression of diabetic retinopathy (DR) using new strategies to obtain genetic information in type 2 diabetes (T2D) based on interfering ribonucleic acid (RNA). MATERIAL AND METHODS: A prospective multicentre case-control study of 132 participants was distributed into: T2D with (+DR) or without (-DR) (T2DG; n = 77), and a control group (CG; n = 55). After an eye examination and personal interview, tears were collected for molecular analysis (expression of microRNAs [miRNAs] (miRCURY(TM) ARN Isolation Kit, Qiagen)]. Libraries, 137 vs. 140 bp (GeneMapper, Applied Biosystems), were obtained in 18 samples (T2DG+DR = 6; T2DG-DR = 6; CG = 6) by performing next-generation sequencing (NGS). SPSS 15.0 statistical program was used to perform data analysis. RESULTS: The mean age was 67 ± 12 years in the T2DG vs. 55 ± 21 years in the CG. Distribution men/women: 25/30 in T2DG vs. 51/28 in CG. A family history of DM, diet compliance, smoking, drinking and exercise, showed significant differences between groups (P<.001). A 20-25 microlitre sample of tears contained a mean of 9.42 ± 3.30 ng/mL of purified ARN, with significant differences between T2DG/CG (P=.002) and T2DG+RD/CG (P=.004). Tear expression of miARNs in T2DG directly correlated with age/obesity/T2D duration (P<.05), and indirectly with visual acuity (P<.05). A total of 14 miRNAs related to the presence, pathogenic mechanisms and risk factors for the progression of diabetic retinopathy, were identified. CONCLUSIONS: We propose to use tears as a source of genetic information for DM. Specific miRNAs involved in DR development and/or progression can be used as molecular biomarkers, and based on these, for developing future biotherapies

Nutrición y Epigenética en Pediatría / Nutrition and Epigenetics in Pediatrics

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Down-regulation of Wnt10a by RNA interference inhibits proliferation and promotes apoptosis in mouse embryonic palatal mesenchymal cells through Wnt/Beta-catenin signaling pathway

Cleft palate is one of the most common birth defects. Both environmental and genetic factors are involved in this disorder. Here, we investigated the function of Wnt10a in proliferation and apoptosis of mouse embryonic palatal mesenchymal (MEPM) cells. Expression of Wnt10a was down-regulated at both the mRNA and protein levels in transfected MEPM cells containing Wnt10a-specific small hairpin RNA (shRNA) plasmid. Down-regulation of Wnt10a inhibited cell proliferation and induced cell cycle arrest in the S phase in MEPM cells. Moreover, apoptosis was significantly increased in MEPM cells of Wnt10a gene silencing. Finally, the expression of β-catenin was markedly reduced in MEPM cells transfected with shRNA plasmid, indicating that the canonical Wnt/β-catenin signaling pathway was involved in the alterations of cell proliferation and apoptosis induced by Wnt10a knockdown. Thus, our findings reveal that Wnt10a regulates proliferation and apoptosis of MEPM cells at least partially through the canonical Wnt/β-catenin signaling pathway

Los Receptores con Silenciador Beta2-Adrenérgicos, reducen la presión intraocular: Un nuevo acercamiento a la terapia del Glaucoma / Silencing Beta 2-Adrenergic Receptors Reduces Intraocular Pressure: A New Approach for Glaucoma Therapy

La búsqueda de nuevos tratamientos para la hipertensión ocular y el glaucoma, dado los efectos secundarios que presentan los fármacos actuales, sugieren el desarrollo de estrategias diferentes a las actualmente disponibles. En este sentido se ha puesto a punto la tecnología de los RNA de interferencia (RNAi) también llamados siRNA para silenciando selectivamente los receptores beta2 adrenérgicos tratar de obtener una reducción sustancial de la presión intraocular (PIO), principal factor desencadenante del glaucoma. El empleo de un siRNA de origen comercial para el receptor beta2 ha producido una reducción sustancial de la PIO de 30 ± 5 %, comparado con el control, un siRNA sin sentido (scramble). Estos resultados fueron en magnitud semejantes al de los fármacos comerciales salvo que la duración del siRNA para el receptor beta2 adrenérgico duró casi 5 días en comparación de las 8 horas que suelen durar los fármacos comerciales. Los estudios en relación con los efectos secundarios no mostraron ninguna modificación en las estructuras oculares. En resumen, el empleo de los siRNA frente a los receptores beta2 adrenérgicos presenta muy buenas perspectivas como aproximación novedosa para el tratamiento del glaucoma(AU)

The search for new treatments for ocular hypertension and glaucoma, due to the side effects commercial products present, is inviting to look for new strategies apart from the available ones. In this sense the interference RNA technique (RNAi), also known as siRNA to selectively silence beta 2 adrenergic receptors has been investigated. In this sense it has been obtained a reduction in intraocular pressure (IOP), the main factor triggering glaucoma, of 30 ± 5 %, compared to a control (scrambled siRNA). The results were in terms of IOP reduction similar to that obtained with commercial compounds but the duration of the siRNA for the beta 2 adrenoceptor lasted for almost 5 days, compared to the average of 8 hours in the case of the other commercial compounds. No apparent side effects were observed in the ocular structures. In summary, the use of siRNA against the beta 2 adrenergic receptors open a new perspective for the treatment of glaucoma(AU)

Double-stranded Let-7 mimics, potential candidates for cancer gene therapy

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MicroRNAs (miRNAs), a class of small, single-stranded endogenous RNAs, act as post-transcriptional regulators of gene expression. The ability of one single miRNA regulating multiple functionally related mRNAs makes it a new potential candidate for cancer gene therapy. Let-7s miRNAs have been demonstrated as tumor-suppressor genes in various types of cancers, providing one choice of gene therapy by replenishing this miRNA. In the present studies, we demonstrate that the chemically synthesized, double-stranded Let-7 mimics can inhibit the growth and migration and induce the cell cycle arrest of lung cancer cell lines in vitro. Let-7 mimics silence gene expression by binding to the 3' UTR of targeting mRNAs. Mutation of seed sequence significantly depresses the gene silencing activity of Let-7 mimics. Our results also demonstrate that it is possible to increase the activity of Let-7s through mutating the sequence within the 3'end of the antisense strand. Directly, co-transfection Let-7 mimics with active siRNAs impairs the anti-cancer activities of Let-7 mimics. However, a 3-h interval between the introduction of Let-7 mimics and a kind of siRNA avoids the competition and enhances the anti-cancer activities of Let-7 mimics. Taken together, these results have revealed that Let-7s mimics are potential candidates for cancer gene therapy (AU)

Inhibición del receptor FGFR3 por ARNs de interferencia para la acondroplasia / Fibroblast growth factor receptor 3 inhibition by small interfering RNAs in achondroplasia

Achondroplasia is a short-limbed dwarfism resulting frommutation and gain-of-function in fibroblast growth factor receptor 3(FGFR3). Effective therapy for this condition has not as yet beenestablished. We have tested the efficiency of three different smallinterference RNAs (siRNAs) to abrogate the FGFR3 expression in humanimmortalized chondrocytes carrying the achondroplasia mutation(G380R). Two siRNA sequences induced markedly decrease of FGFR3mRNA (up to 75% reduction) and protein levels (up to 61% reduction).Furthemore, siRNA-mediated knockdown of FGFR3 blocked theactivation of the downstream signal transduction ERK pathway(AU)

Inhibición del receptor FGFR3 por ARNs de interferencia para laacondroplasiaLa acondroplasia es un tipo de enanismo caracterizado porextremidades cortas resultante de una mutación en el receptor decrecimiento de fibroblastos de tipo 3 (FGFR3). Aún no se ha establecidouna terapia efectiva para esta enfermedad. Nosotros hemos testado laeficiencia de tres diferentes small interference RNAs (siRNAs) parabloquear la expresión del receptor FGFR3 en condrocitos humanosinmortalizados portadores de la mutación acondroplásica (G380R). Dossecuencias de siRNAs indujeron un marcado descenso de la expresión deARN mensajero del receptor FGFR3 (hasta un 75%) así como de losniveles de proteína (hasta un 61%). Además, el bloqueo de la expresióndel receptor FGFR3 mediado por los siRNAs redujo la activación de lacascada de transducción de las ERK(AU)

ARN de interferencia terapéutico para enfermedades neurodegenerativas / Therapeutic RNA interference for neurodegenerative diseases

Introducción. El descubrimiento del ARN de interferencia (ARNi), una vía biológica de control de expresión génica,ha revolucionado el campo de la investigación biomédica. De las muchas aplicaciones del ARNi, quizás su explotación terapéutica sea la más prometedora. Un grupo de enfermedades donde el uso de ARNi terapéutico se está explorando activamente lo constituyen los trastornos neurodegenerativos. Objetivo. Presentar tanto al neurólogo clínico como al investigador básico un sumario actualizado del desarrollo de ARNi terapéutico para enfermedades neurodegenerativas. Desarrollo. El progresotécnico en la manipulación del ARNi en neuronas y los avances recientes en el conocimiento de la patogenia de los procesos neurodegenerativos han facilitado el diseño de ARNi terapéutico tanto para enfermedades hereditarias como idiopáticas. Varios ensayos preclínicos se han completado con éxito en modelos animales, allanando el camino hacia el diseño de ensayos clínicos en humanos. Sin embargo, antes de llegar a ese punto hay que completar más estudios experimentales en animalespara demostrar la eficacia de esta modalidad terapéutica y, más importante aún, para predecir los posibles efectos adversos de esta intervención en el cerebro humano. Conclusión. Aunque todavía en estadios preclínicos, la explotación del ARNi con fines terapéuticos en enfermedades neurodegenerativas está generando resultados muy prometedores. La evoluciónde este campo en los próximos años puede ser crítica para iniciar su aplicación clínica

Introduction. The discovery of RNA interference (RNAi), a biological way to control gene expression, has revolutionised the field of biomedical research. Of the many applications of RNAi, perhaps its use for therapeutic purposes is the most promising. One group of diseases where the use of therapeutic RNAi is being actively explored is that of neurodegenerativedisorders. Aim. To provide both the clinical neurologist and the basic researcher with an updated summary of the development of therapeutic RNAi for neurodegenerative diseases. Development. The technical progress made in the manipulation of RNAi in neurons and the recent advances in our knowledge of the pathogenesis of neurodegenerative processes have been valuableaids in designing therapeutic RNAi for both hereditary and idiopathic diseases. Several pre-clinical trials have been successfully completed in animal models, thus clearing the way towards the design of clinical trials in humans. Nevertheless, before reaching that point more experimental studies need to be carried out in animals to prove the effectiveness of this mode of therapy and, still more important, to be able to predict the possible side effects of this procedure in the human brain.Conclusions. Although it is still in the pre-clinical stages, the use of RNAi for therapeutic purposes in neurodegenerative diseases is producing some very promising results. The evolution of this field over the next few years may be critical for beginning its clinical application

An easy, rapid and objective mathematical method to identify fatty acid synthase (oncogenic antigen-519) modulators with potential anticancer value

Fatty acid synthase (FASN) is a novel druggable target for metabolically treating and preventing human malignancies. We envisioned that if loss of sensitivity to C75 (a slow-binding FASN inhibitor) occurs in parallel with loss of FASN expression and/or activity, a mathematical assessment of the nature of the interaction between investigational FASN modulators and C75 may predict the ability of experimental compounds to regulate FASN. We statistically compared the arithmetical sums of the anti-proliferative effects obtained when FASN modulators and C75 were used as single agents to those observed experimentally when agents were actually combined in a sequential schedule (i.e., FASN modulator-->C75). A reduced sensitivity to C75 (antagonism) occurred when compounds down-regulated FASN activity/expression, while an enhanced C75 efficacy (synergism) was found following exposure to FASN up-regulators. This "C75-sensitivity test" might offer an easy, rapid and objective method to identify FASN inhibitors with potential anticancer value in human cancer (AU)

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El ácido ribonucleico de interferencia. ¿Una curiosidad que merece un premio Nobel o una esperanza terapéutica? / Interference RNA: a curiosity that merits a nobel price or a future therapeutic tool

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Terapias basadas en la tecnología de ARN de interferencia: una nueva clase de fármacos para el tratamiento de patologías oculares / RNA interferences therapeutics: a new approach to the treatment of ocular diseases

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microRNAs and cancer: role in tumorigenesis, patient classification and therapy

MicroRNAs (miRNAs) are small non-coding RNAs that downregulate gene expression during various crucial cell processes such as apoptosis, differentiation and development. Recent work supports a role for miRNAs in the initiation and progression of human malignancies. Moreover, large high-throughput studies in patients revealed that miRNA profiling has the potential to classify tumours and predict patient outcome with high accuracy. Functional studies, some of which involve animal models, indicate that miRNAs act as tumour suppressors and oncogenes. This review examines the role of miRNAs in the pathogenesis of cancer as well as miRNA-profiling studies performed in human malignancies. Implications of these findings for the diagnosis and treatment of cancer patients are also discussed (AU)

Terapia génica en neurooncología / Gene therapy in neuro-oncology

Introducción y desarrollo. El concepto de terapia génicaimplica la transferencia de material genético a una célula, un tejidoo un órgano con el fin de curar una enfermedad o al menosmejorar el estado clínico de un paciente. Los componentes esencialesde la terapia son: un vector capaz de introducirse eficiente yselectivamente en las células que constituyan el foco de la enfermedad,y un gen terapéutico que represente un remedio genético parala patología. En terapias contra el cáncer se utilizan preferentementelos retrovirus como vectores al infectar exclusivamente lascélulas en división. Entre las estrategias más populares y exitosasde terapia génica contra el cáncer se encuentran la inmunoterapia,la antiangiogénesis, la utilización de genes ‘asesino-suicidas’ yel bloqueo de oncogenes con ARN de interferencia. Conclusiones.Cada una de estas terapias se ha verificado en líneas tumoralescultivadas, así como en animales de experimentación, donde se haobservado un efecto antitumoral claro con ausencia de efectossecundarios apreciables. En la actualidad, la mayoría de estas estrategiasse encuentran en fase clínica, pero aún no constituyenuna opción terapéutica

Introduction and development. The concept of gene therapy implies the transfer of genetic material to a cell, a tissueor an organ with the aim to cure an illness or at least to improve the clinical status of a patient. The essential components ofthe therapy are: a vector able to enter efficiently and selectively the target cells; and the therapeutic gene competent to curethe pathology. In cancer therapies, retroviruses are preferentially used as vectors because they only infect dividing cells.Among the most popular and successful gene therapies against cancer we find the immunotherapy, the antiangiogenicstrategies, the killer-suicide genes and RNA interference against specific oncogenes. Conclusions. Each one of these therapieshave proven successful in cancer cell lines and in experimental animals where it has been shown a clear antitumoral effect inthe absence of appreciable secondary effects. At present, the majority of these procedures are in clinical phases but none as yetis considered a therapeutic option