A Novel and Efficient Gene Transfer Strategy Reduces Glial Reactivity and Improves Neuronal Survival and Axonal Growth In Vitro

Background: The lack of axonal regeneration in the central nervous system is attributed among other factors to the formation of a glial scar. This cellular structure is mainly composed of reactive astrocytes that overexpress two intermediate filament proteins, the glial fibrillary acidic protein (GFAP) and vimentin. Indeed, in vitro, astrocytes lacking GFAP or both GFAP and vimentin were shown to be the substrate for increased neuronal plasticity. Moreover, double knockout mice lacking both GFAP and vimentin presented lower levels of glial reactivity in vivo, significant axonal regrowth and improved functional recovery in comparison with wild-type mice after spinal cord hemisection. From these results, our objective was to develop a novel therapeutic strategy for axonal regeneration, based on the targeted suppression of astroglial reactivity and scarring by lentiviral-mediated RNA-interference (RNAi). Methods and Findings: In this study, we constructed two lentiviral vectors, Lv-shGFAP and Lv-shVIM, which allow efficient and stable RNAi-mediated silencing of endogenous GFAP or vimentin in vitro. In cultured cortical and spinal reactive astrocytes, the use of these vectors resulted in a specific, stable and highly significant decrease in the corresponding protein levels. In a second model -scratched primary cultured astrocytes- Lv-shGFAP, alone or associated with Lv-shVIM, decreased astrocytic reactivity and glial scarring. Finally, in a heterotopic coculture model, cortical neurons displayed higher survival rates and increased neurite growth when cultured with astrocytes in which GFAP and vimentin had been invalidated by lentiviral-mediated RNAi. Conclusions: Lentiviral-mediated knockdown of GFAP and vimentin in astrocytes show that GFAP is a key target for modulating reactive gliosis and monitoring neuron/glia interactions. Thus, manipulation of reactive astrocytes with the Lv-shGFAP vector constitutes a promising therapeutic strategy for increasing glial permissiveness and permitting axonal regeneration after central nervous system lesions. Copyright: © 2009 Desclaux et al.This work was supported by the Centre National de la Recherche Scientifique (CNRS), the Institut National de la Sante et de la Recherche Medicale (INSERM), the Universite Pierre et Marie Curie - Paris 6 (UPMC), the Universite de Montpellier 2, Verticale, Demain Debout, and the Institut de Recherche sur la Moelle epiniere et l’Encephale (IRME).Peer Reviewe

The Educational Robotics: a resource to enhance the scientific and technological capabilities

La caracterización de la sociedad actual supone analizarla desde diversas perspectivas, la social, la política, la ética, la económica, la cultural y la educativa. En el ámbito educativo diferentes autores hacen referencia a la necesidad de revisar las prácticas educativas para responder a esas caracterizaciones sociales, que requiere de individuos con capacidades para poder insertarse y desempeñarse como ciudadanos de este siglo. El modelo tradicional de transmisión de conocimientos y adquisición de contenidos deberían dar paso a la adquisición y desarrollo de capacidades que habiliten a los estudiantes para afrontar los desafíos de estos nuevos contextos y escenarios. La introducción de La Robótica Educativa en el aula, es un recurso y una estrategia innovadora que propicia la integración de contenidos intra e interdisciplinar, pone en evidencia la relación teoría-práctica, desarrolla la curiosidad, la exploración y la creatividad; y fomenta el desarrollo de capacidades fundamentales y científico-tecnológicas; el siguiente documento expone los resultados obtenidos de la aplicación de esta herramienta.Tema: Experiencia Docente.Red de Universidades con Carreras en Informátic

The Educational Robotics: a resource to enhance the scientific and technological capabilities

La caracterización de la sociedad actual supone analizarla desde diversas perspectivas, la social, la política, la ética, la económica, la cultural y la educativa. En el ámbito educativo diferentes autores hacen referencia a la necesidad de revisar las prácticas educativas para responder a esas caracterizaciones sociales, que requiere de individuos con capacidades para poder insertarse y desempeñarse como ciudadanos de este siglo. El modelo tradicional de transmisión de conocimientos y adquisición de contenidos deberían dar paso a la adquisición y desarrollo de capacidades que habiliten a los estudiantes para afrontar los desafíos de estos nuevos contextos y escenarios. La introducción de La Robótica Educativa en el aula, es un recurso y una estrategia innovadora que propicia la integración de contenidos intra e interdisciplinar, pone en evidencia la relación teoría-práctica, desarrolla la curiosidad, la exploración y la creatividad; y fomenta el desarrollo de capacidades fundamentales y científico-tecnológicas; el siguiente documento expone los resultados obtenidos de la aplicación de esta herramienta.Tema: Experiencia Docente.Red de Universidades con Carreras en Informátic

The Educational Robotics: a resource to enhance the scientific and technological capabilities

La caracterización de la sociedad actual supone analizarla desde diversas perspectivas, la social, la política, la ética, la económica, la cultural y la educativa. En el ámbito educativo diferentes autores hacen referencia a la necesidad de revisar las prácticas educativas para responder a esas caracterizaciones sociales, que requiere de individuos con capacidades para poder insertarse y desempeñarse como ciudadanos de este siglo. El modelo tradicional de transmisión de conocimientos y adquisición de contenidos deberían dar paso a la adquisición y desarrollo de capacidades que habiliten a los estudiantes para afrontar los desafíos de estos nuevos contextos y escenarios. La introducción de La Robótica Educativa en el aula, es un recurso y una estrategia innovadora que propicia la integración de contenidos intra e interdisciplinar, pone en evidencia la relación teoría-práctica, desarrolla la curiosidad, la exploración y la creatividad; y fomenta el desarrollo de capacidades fundamentales y científico-tecnológicas; el siguiente documento expone los resultados obtenidos de la aplicación de esta herramienta.Tema: Experiencia Docente.Red de Universidades con Carreras en Informátic

Array-Comparative Genomic Hybridization Analysis in Fetuses with Major Congenital Malformations Reveals that 24% of Cases Have Pathogenic Deletions/Duplications

Karyotyping and aCGH are routinely used to identify genetic determinants of major congenital malformations (MCMs) in fetal deaths or terminations of pregnancy after prenatal diagnosis. Pathogenic rearrangements are found with a variable rate of 9-39% for aCGH. We collected 33 fetuses, 9 with a single MCM and 24 with MCMs involving 2-4 organ systems. aCGH revealed copy number variants in 14 out of 33 cases (42%). Eight were classified as pathogenic which account for a detection rate of 24% (8/33) considering fetuses with 1 or more MCMs and 33% (8/24) taking into account fetuses with multiple malformations only. Three of the pathogenic variants were known microdeletion syndromes (22q11.21 deletion, central chromosome 22q11.21 deletion, and TAR syndrome) and 5 were large rearrangements, adding up to >11 Mb per subject and comprising strong phenotype-related genes. One of those was a de novo complex rearrangement, and the remaining 4 duplications and 2 deletions were 130-900 kb in size, containing 1-7 genes, and were classified as variants of unknown clinical significance. Our study confirms aCGH as a powerful technique to ascertain the genetic etiology of fetal major congenital malformations

PIK3R1W624R Is an Actionable Mutation in High Grade Serous Ovarian Carcinoma.

Identifying cancer drivers and actionable mutations is critical for precision oncology. In epithelial ovarian cancer (EOC) the majority of mutations lack biological or clinical validation. We fully characterized 43 lines of Patient-Derived Xenografts (PDXs) and performed copy number analysis and whole exome sequencing of 12 lines derived from naïve, high grade EOCs. Pyrosequencing allowed quantifying mutations in the source tumours. Drug response was assayed on PDX Derived Tumour Cells (PDTCs) and in vivo on PDXs. We identified a PIK3R1W624R variant in PDXs from a high grade serous EOC. Allele frequencies of PIK3R1W624R in all the passaged PDXs and in samples of the source tumour suggested that it was truncal and thus possibly a driver mutation. After inconclusive results in silico analyses, PDTCs and PDXs allowed the showing actionability of PIK3R1W624R and addiction of PIK3R1W624R carrying cells to inhibitors of the PI3K/AKT/mTOR pathway. It is noteworthy that PIK3R1 encodes the p85α regulatory subunit of PI3K, that is very rarely mutated in EOC. The PIK3R1W624R mutation is located in the cSH2 domain of the p85α that has never been involved in oncogenesis. These data show that patient-derived models are irreplaceable in their role of unveiling unpredicted driver and actionable variants in advanced ovarian cancer

Cutaneous squamous cell carcinoma with mucinous metaplasia on the sole associated with high-risk human papillomavirus type 18

A case of superficially invasive cutaneous squamous cell carcinoma (SCC) of the sole containing numerous mucin-producing vacuolated cells resembling "signet-ring" cells is reported. The 2 cellular components of the tumor, both squamous and mucinous, were atypical with pleomorphic nuclei, and expressed the same immunophenotype, consistent in weak and focal positivity for cytokeratin 5/6 and epithelial membrane antigen (EMA) and weak cytoplasmic and nuclear positivity for p16. Real-time PCR genotyping demonstrated the presence of high-risk human papillomavirus (HPV) type 18. We diagnose our case as "cutaneous SCC with mucinous metaplasia" and discuss the differential diagnoses with other skin tumors exhibiting mucin-containing cells, in particular with adenosquamous carcinoma and mucoepidermoid carcinoma. Although HPV 18 is not uncommon in cervico-vaginal pathology, where is often associated with mucinous adenocarcinoma or adenosquamous carcinoma of the cervix, its detection has been rarely reported in cutaneous SCC. In our case, the association of mucinous metaplasia and oncogenic high-risk HPV 18 in a cutaneous SCC may be of interest to the dermatopathologist. Further observations need to confirm whether the histopathologic finding of mucinous metaplasia in an atypical squamous cell proliferation could be a clue for investigating the presence of oncogenic high-risk HPV infection, with particular regard to HPV 18 subtype