Studi pre-clinici per lo sviluppo di approcci terapeutici basati sui microRNA in un modello di Leucemia Linfatica Cronica

Despite its indolent nature, chronic lymphocytic leukemia (CLL) is an incurable disease. The involvement of microRNAs (miRNAs) in CLL pathogenesis has been established and it suggests that miRNAs are attractive candidates as therapeutic targets for CLL. Here, we employed the Eμ-TCL1 transgenic mouse model, which develops a chronic B-cell CD5+ leukemia similar to an aggressive form of human B-CLLs, to investigate miR-181b and miR-34a as a new therapeutic agents. In vitro, the enforced expression of miR-181b mimics induced a significant apoptotic effect in human B-cells (RAJI, DAUDI, 697, EHEB) as well as in mouse Eμ-TCL1 leukemic splenocytes. In addition molecular analyses revealed that miR-181b not only affected the expression of Tcl1, but also of Bcl2 and Mcl1 anti-apoptotic proteins, and reduced the phosphorylation levels of Akt and Erk1/2. Similarly, the restoration of miR-34a induced an increase of apoptosis in murine leukemic spleen cells, which could be related to the modulation of CDK6 and Bcl-2 targets. Finally, in vivo multiple treatments with miR-181b or miR-34a mimics induced a reduction/delay in the progression of leukemia and an increase of overall survival in treated mice. Our pre-clinical assessment of miRNA-based therapies in a CLL mouse model proves a sizable anti-leukemic activity of miR-181b and miRNA-34a as single agents, and suggests that the use of microRNAs could represent a novel potential therapeutic approach for the treatment of CLL

Midiendo la influencia del sector empresario en las políticas públicas : un análisis de redes sociales de la legislación económica

El objetivo de este estudio fue investigar el proceso de toma de decisiones legislativas en el área económica durante la última dictadura argentina (1976-1983) utilizando los archivos desclasificados de la Comisión de Asesoramiento Legislativo. Para ello se analizó en profundidad a los actores y sus interacciones con la metodología de redes sociales. El estudio se dividió en tres etapas. En una primera instancia se llevó a cabo una revisión teórica. Posteriormente se recolectó y procesó toda la información y por último, se construyeron redes sociales utilizando un software especializado y se analizaron los esquemas. A partir de las interacciones visibles en las redes sociales fue posible extraer algunas conclusiones sobre los distintos actores que estuvieron involucrados en mayor o menor medida en el proceso de toma de decisiones en materia de legislación económica. El trabajo se centró en las presidencias de Viola y Galtieri y examinó la pertenencia de los actores, la participación del sector público y privado, el origen de las interacciones y la influencia de los sectores transables y no transables

Awareness of Social Presence on Virtual Fitness Platforms and Relationship with Exercise Motivation and Physical Activity Levels

The health benefits of physical activity are well-known, however, only 20% of the U.S. population meets the Physical Activity Guidelines for Americans. This study aimed to explore the association of awareness of social presence in a virtual fitness platform with motivation and physical activity levels (PAL). Virtual fitness users (n = 590, 42 ± 12.7 years old) completed the International Physical Activity Questionnaire and Behavioral Regulation of Exercise Questionnaire. Relative autonomy was correlated with PAL (r = .21, p < .001, 95.00% CI = [.13, .29]) and predicted PAL (F(1,588) = 27.03, p < .001). Awareness of social presence was significantly related to motivation (U = 41864.5, z = –5.99, p < .001), and predictive of relative autonomy (F(1,588) = 27.03, p < .001). The results suggest that higher relative autonomy is associated with higher PAL in virtual fitness users. Awareness of social presence on virtual platform appears to correlate to higher levels of relative autonomy, which may influence exercise adherence

Sopra alcuni particolari effetti delle proiezioni cinematografiche nei nevrotici (1911)

Da parecchi anni a questa parte la mia attenzione è stata attirata sopra alcune determinate turbe nervose, che possono rilevarsi nei nevrotici in seguito all’avere assistito a particolari rappresentazioni cinematografiche. Nessun dubbio sul godimento intellettuale, che possono offrire le projezioni cinematografiche; esse costituiscono una gradita distrazione, e possono riuscire di grande istruzione, per cui il pubblico d’ogni condizione ed età, dal fanciullo al vecchio, dall’analfabeta all’u..

Anti-leukemic activity of microRNA-26a in a chronic lymphocytic leukemia mouse model

Dysregulation of microRNAs (miRNAs) plays an important role in the pathogenesis of chronic lymphocytic leukemia (CLL). The Emu-TCL1 transgenic mouse develops a form of leukemia that is similar to the aggressive type of human B-CLL, and this valuable model has been widely used for testing novel therapeutic approaches. Here, we adopted this model to investigate the potential effects of miR-26a, miR-130an and antimiR-155 in CLL therapy. Improved delivery of miRNA molecules into CLL cells was obtained by developing a novel system based on lipid nanoparticles conjugated with an anti-CD38 monoclonal antibody. This methodology has proven to be highly effective in delivering miRNA molecules into leukemic cells. Short- and long-term experiments showed that miR-26a, miR-130a and anti-miR-155 increased apoptosis after in vitro and in vivo treatment. Of this miRNA panel, miR-26a was the most effective in reducing leukemic cell expansion. Following long-term treatment, apoptosis was readily detectable by analyzing cleavage of PARP and caspase-7. These effects could be directly attributed to miR-26a, as confirmed by significant downregulation of its proven targets, namely cyclin-dependent kinase 6 and Mcl1. The results of this study are relevant to two distinct areas. The first is related to the design of a technical strategy and to the selection of CD38 as a molecular target on CLL cells, both consenting efficient and specific intracellular transfer of miRNA. The original scientific finding inferred from the above approach is that miR-26a can elicit in vivo anti-leukemic activities mediated by increased apoptosi

miR-125b targets erythropoietin and its receptor and their expression correlates with metastatic potential and ERBB2/HER2 expression

Background The microRNA 125b is a double-faced gene expression regulator described both as a tumor suppressor gene (in solid tumors) and an oncogene (in hematologic malignancies). In human breast cancer, it is one of the most down-regulated miRNAs and is able to modulate ERBB2/3 expression. Here, we investigated its targets in breast cancer cell lines after miRNA-mimic transfection. We examined the interactions of the validated targets with ERBB2 oncogene and the correlation of miR-125b expression with clinical variables. Methods MiR-125b possible targets were identified after transfecting a miRNA-mimic in MCF7 cell line and analyzing gene expression modifications with Agilent microarrays and Sylamer bioinformatic tool. Erythropoietin (EPO) and its receptor (EPOR) were validated as targets of miR-125b by luciferase assay and their expression was assessed by RT-qPCR in 42 breast cancers and 13 normal samples. The molecular talk between EPOR and ERBB2 transcripts, through miR-125b, was explored transfecting MDA-MD-453 and MDA-MB-157 with ERBB2 RNA and using RT-qPCR. Results We identified a panel of genes down-regulated after miR-125b transfection and putative targets of miR-125b. Among them, we validated erythropoietin (EPO) and its receptor (EPOR) - frequently overexpressed in breast cancer - as true targets of miR-125b. Moreover, we explored possible correlations with clinical variables and we found a down-regulation of miR-125b in metastatic breast cancers and a significant positive correlation between EPOR and ERBB2/HER2 levels, that are both targets of miR-125b and function as competing endogenous RNAs (ceRNAs). Conclusions Taken together our results show a mechanism for EPO/EPOR and ERBB2 co-regulation in breast cancer and confirm the importance of miR-125b in controlling clinically-relevant cancer features

Anti-Tumor Activity of a miR-199-dependent Oncolytic Adenovirus

none15siThe down-regulation of miR-199 occurs in nearly all primary hepatocellular carcinomas (HCCs) and HCC cell lines in comparison with normal liver. We exploited this miR-199 differential expression to develop a conditionally replication-competent oncolytic adenovirus, Ad-199T, and achieve tumor-specific viral expression and replication. To this aim, we introduced four copies of miR-199 target sites within the 3' UTR of E1A gene, essential for viral replication. As consequence, E1A expression from Ad-199T virus was tightly regulated both at RNA and protein levels in HCC derived cell lines, and replication controlled by the level of miR-199 expression. Various approaches were used to asses in vivo properties of Ad-199T. Ad-199T replication was inhibited in normal, miR-199 positive, liver parenchyma, thus resulting in reduced hepatotoxicity. Conversely, the intrahepatic delivery of Ad-199T in newborn mice led to virus replication and fast removal of implanted HepG2 liver cancer cells. The ability of Ad-199T to control tumor growth was also shown in a subcutaneous xenograft model in nude mice and in HCCs arising in immune-competent mice. In summary, we developed a novel oncolytic adenovirus, Ad-199T, which could demonstrate a therapeutic potential against liver cancer without causing significant hepatotoxicityopenCallegari E; Elamin BK; D'Abundo L; Falzoni S; Donvito G; Moshiri F; Milazzo M; Altavilla G; Giacomelli L; Fornari F; Hemminki A; Di Virgilio F; Gramantieri L; Negrini M; Sabbioni SCallegari E; Elamin BK; D'Abundo L; Falzoni S; Donvito G; Moshiri F; Milazzo M; Altavilla G; Giacomelli L; Fornari F; Hemminki A; Di Virgilio F; Gramantieri L; Negrini M; Sabbioni