Web "Geologia urbana de Badalona". Les seves aplicacions a l'aula

La web "Geologia Urbana de Badalona" planteja l'estudi de les roques i els seus elements constituents mitjançant recorreguts urbans i tenint com a objecte els materials geològics ornamentals emprats en construcció. Aquesta web s'ha concebut, per una banda, com a informació per a una possible alternativa a les sortides de camp, que són costoses i solen generar problemes de calendari i, per una altra, amb la idea de complementar amb el contingut de la pròpia web les sortides als carrers i places propers al centre escolar, on sovint hi ha una bona representació dels tipus de roques més habituals

Applying Microsatellite Multiplex PCR Analysis (MMPA) for Determining Allele Copy-Number Status and Percentage of Normal Cells within Tumors

The study of somatic genetic alterations in tumors contributes to the understanding and management of cancer. Genetic alterations, such us copy number or copy neutral changes, generate allelic imbalances (AIs) that can be determined using polymorphic markers. Here we report the development of a simple set of calculations for analyzing microsatellite multiplex PCR data from control-tumor pairs that allows us to obtain accurate information not only regarding the AI status of tumors, but also the percentage of tumor-infiltrating normal cells, the locus copy-number status and the mechanism involved in AI. We validated this new approach by re-analyzing a set of Neurofibromatosis type 1-associated dermal neurofibromas and comparing newly generated data with results obtained for the same tumors in a previous study using MLPA, Paralog Ratio Analysis and SNP-array techniques. Microsatellite multiplex PCR analysis (MMPA) should be particularly useful for analyzing specific regions of the genome containing tumor suppressor genes and also for determining the percentage of infiltrating normal cells within tumors allowing them to be sorted before they are analyzed by more expensive techniques

A novel long non-coding RNA from NBL2 pericentromeric macrosatellite forms a perinucleolar aggregate structure in colon cancer

Primate-specific NBL2 macrosatellite is hypomethylated in several types of tumors, yet the consequences of this DNA hypomethylation remain unknown. We show that NBL2 conserved repeats are close to the centromeres of most acrocentric chromosomes. NBL2 associates with the perinucleolar region and undergoes severe demethylation in a subset of colorectal cancer (CRC). Upon DNA hypomethylation and histone acetylation, NBL2 repeats are transcribed in tumor cell lines and primary CRCs. NBL2 monomers exhibit promoter activity, and are contained within novel, non-polyA antisense lncRNAs, which we designated TNBL (Tumor-associated NBL2 transcript). TNBL is stable throughout the mitotic cycle, and in interphase nuclei preferentially forms a perinucleolar aggregate in the proximity of a subset of NBL2 loci. TNBL aggregates interact with the SAM68 perinucleolar body in a mirror-image cancer specific perinucleolar structure. TNBL binds with high affinity to several proteins involved in nuclear functions and RNA metabolism, such as CELF1 and NPM1. Our data unveil novel DNA and RNA structural features of a non-coding macrosatellite frequently altered in cancer

Using antisense oligonucleotides for the physiological modulation of the alternative splicing of NF1 exon 23a during PC12 neuronal differentiation

Neurofibromatosis Type 1 (NF1) is a genetic condition affecting approximately 1:3500 persons worldwide. The NF1 gene codes for neurofibromin protein, a GTPase activating protein (GAP) and a negative regulator of RAS. The NF1 gene undergoes alternative splicing of exon 23a (E23a) that codes for 21 amino acids placed at the center of the GAP related domain (GRD). E23a-containing type II neurofibromin exhibits a weaker Ras-GAP activity compared to E23a-less type I isoform. Exon E23a has been related with the cognitive impairment present in NF1 individuals. We designed antisense Phosphorodiamidate Morpholino Oligomers (PMOs) to modulate E23a alternative splicing at physiological conditions of gene expression and tested their impact during PC12 cell line neuronal differentiation. Results show that any dynamic modification of the natural ratio between type I and type II isoforms disturbed neuronal differentiation, altering the proper formation of neurites and deregulating both the MAPK/ERK and cAMP/PKA signaling pathways. Our results suggest an opposite regulation of these pathways by neurofibromin and the possible existence of a feedback loop sensing neurofibromin-related signaling. The present work illustrates the utility of PMOs to study alternative splicing that could be applied to other alternatively spliced genes in vitro and in vivo

Reprogramming Captures the Genetic and Tumorigenic Properties of Neurofibromatosis Type 1 Plexiform Neurofibromas

Neurofibromatosis type 1 (NF1) is a tumor predisposition genetic disease caused by mutations in the NF1 tumor suppressor gene. Plexiform neurofibromas (PNFs) are benign Schwann cell (SC) tumors of the peripheral nerve sheath that develop through NF1 inactivation and can progress toward a malignant soft tissue sarcoma. There is a lack of non-perishable model systems to investigate PNF development. We reprogrammed PNF-derived NF1(-/-) cells, descendants from the tumor originating cell. These NF1(-/-)-induced pluripotent stem cells (iPSCs) captured the genomic status of PNFs and were able to differentiate toward neural crest stem cells and further to SCs. iPSC-derived NF1(-/-) SCs exhibited a continuous high proliferation rate, poor myelination ability, and a tendency to form 3D spheres that expressed the same markers as their PNF-derived primary SC counterparts. They represent a valuable model to study and treat PNFs. PNF-derived iPSC lines were banked for making them available

Using Phosphorodiamidate Morpholino Oligomers (PMOs) to characterize the role of neurofibromin in cell physiology

[cat] La neurofibromina és el producte del gen NF1, que mutat causa la Neurofibromatosis de tipus 1. Tot i que en l'actualitat, encara ens cal entendre millor el rol d'aquesta proteïna en la fisiologia cel•lular, l'activitat Ras-GAP és la funció bioquímica més ben caracteritzada de la neurofibromina. Aquesta funció està regulada per l'splicing alternatiu de l'exó 23a (E23a). En aquesta tesi ens vàrem proposar comprendre millor el paper d'aquest splicing alternatiu durant el procés de diferenciació neuronal, amb l'objectiu de proporcionar nova informació sobre els problemes cognitius i d'aprenentatge associats a aquesta malaltia. Degut a la gran grandària de la neurofibromina i a la dificultat de manipular-la in vitxo, es varen utilitzar Phosphorodiamidate Morpholino Oligomers (PMOs) per modificar la composició exònica del mARN (per tant l'estructura resultant de la neurofibromina) sense alterar les condicions fisiològiques d'expressió del gen NF 1 . Es va desenvolupar un sistema basat en PMOs per forçar l'expressió de l'isoforma tipus II (+E23a) o tipus I (-E23a) del gen NF1 en cèl•lules PC12, un model de diferenciació neuronal, en presència o absència de Nerve Growth Factor (NGF). A més, per entendre la importància de 1'E23a es va establir un grup de metodologies i assajos funcionals per poder determinar diferents respostes cel•lulars i valorar la funció d'aquest en el procés de diferenciació neuronal. Els nostres resultats van mostrar que forçar l'isoforma tipus I (-E23a) no era suficient per induir la diferenciació de les cèl•lules PC12 en absència de NGF. No obstant això, qualsevol alteració en la relació entre les isoformes tipus I/II en presència de NGF, ja sigui d'una manera quantitativa o dependent del temps, interferia en el correcte procés de diferenciació neuronal, en particular, alterant la correcta formació de neurites, així com l'adequada regulació de les vies de senyalització RAS/MAPK i cAMP/PKA. En conjunt, els resultats d'aquesta tesi indiquen que la regulació de l'splicing alternatiu de 1'E23a del gen NF1 permet un ajust fi de les vies RAS/MAPK i cAMP/PKA a través de la activitat GAP de la neurofibromina, d'una forma oposada i coordinada al llarg del temps durant el procés de diferenciació neuronal.[eng] The role of neurofibromin, the product of the Neurofibromatosis Type 1 (NF1) gene, in cell physiology is still not well understood. Considering the different traits associated to NF1 it is clear that neurofibromin participates in processes of proliferation and differentiation of different cell types. The Ras-GAP activity of neurofibromin is its best characterized biochemical function, which is regulated by the alternative splicing of exon 23a (E23a). In this thesis we intended to better understand the role of alternative splicing of E23a during neuronal differentiation, with the aim that in the future it could provide information on the learning and cognitive issues related to this disease. Due to the large size of neurofibromin, the difficulty of manipulating it in vitro and the necessity to mimic, as much as possible, the physiological conditions of the cell, we used Phosphorodiamidate Morpholino Oligomers (PMOs) to modify the exonic composition of NF1 mRNA while preserving the endogenous neurofibromin expression levels. We developed a PMO-based system that successfully allowed to force the expression of type II (+E23a) or type I (-E23a) isoforms without altering the physiological expression levels of NF1 mRNA in PC12 cells, a neuronal differentiation model, in the presence or absence of Nerve Growth Factor (NGF). This PMO system could be used in other cellular models of NF1, and could be reproduced for studying the regulation of the alternative splicing in other genes. Furthermore, we set up a group of functional assays for assessing proliferation, differentiation, signaling and apoptosis in this PC12 neuronal model. Our results showed that forcing type I (-E23a) isoform was not sufficient for inducing PC12 neuronal differentiation in the absence of NGF. However, the results also demonstrate that any alteration of the NGF-induced ratio between type I/II isoforms, either in a quantitative or time-dependent manner, interfered with the correct neuronal differentiation process, in particular, altering the correct formation of neurites, as well as the proper regulation of the RAS/MAPK and cAMP/PKA signalling pathways. Moreover, the alteration of the natural E23a alternative splicing also impeded the proper neuronal differentiation process in other neuronal models, such as the H19-7 hippocampus cells. Our results also showed that the depletion of neurofibromin in PC12 induce a generalized process of apoptosis; suggest the existence of an early negative feed-back on the function of neurofibromin when type I isoform is abnormally expressed, and shows that the regulation of the cAMP/PKA pathway is also dependent on the GAP domain of neurofibromin. All together, the results of this thesis indicate that the regulation of the alternative splicing of exon 23a of the NF1 gene allows the fine-tuning of the RAS/MAPK and cAMP/PKA pathways through its GAP activity in a coordinate and opposite way along the time-dependent process of neuronal differentiation

Web “Geologia urbana de Badalona”: les seves aplicacions a l’aula

La web “Geologia Urbana de Badalona” planteja l’estudi de les roques i els seus elements constituents mitjançant recorreguts urbans i tenint com a objecte els materials geològics ornamentals emprats en construcció. Aquesta web s’ha concebut, per una banda, com a informació per a una possible alternativa a les sortides de camp, que són costoses i solen generar problemes de calendari i, per una altra, amb la idea de complementar amb el con-tingut de la pròpia web les sortides als carrers i places propers al centre escolar, on sovint hi ha una bona representació dels tipus de roques més habitual

Web "Geologia urbana de Badalona". Les seves aplicacions a l'aula

La web "Geologia Urbana de Badalona" planteja l'estudi de les roques i els seus elements constituents mitjançant recorreguts urbans i tenint com a objecte els materials geològics ornamentals emprats en construcció. Aquesta web s'ha concebut, per una banda, com a informació per a una possible alternativa a les sortides de camp, que són costoses i solen generar problemes de calendari i, per una altra, amb la idea de complementar amb el contingut de la pròpia web les sortides als carrers i places propers al centre escolar, on sovint hi ha una bona representació dels tipus de roques més habituals

Guix

Resumen de los autores en catalánLa noche del diez de mayo, en la playa de Badalona, se quema el demonio. Esta celebración es uno de los ejes fundamentales de las Fiestas de Mayo, y, sin duda, una fecha esperada por todos los niños. El artículo que presentamos es la plasmación de la experiencia llevada a cabo con alumnos de primaria, en colaboración con el CRP de la zona.CataluñaES

Barcelona conference on epigenetics and cancer 2016 – beyond cancer genomes

The Barcelona Conference on Epigenetics and Cancer (BCEC) entitled “Beyond Cancer Genomes” took place October 13th and 14th 2016 in Barcelona. The 2016 BCEC was the fourth edition of a series of annual conferences coordinated by Marcus Buschbeck and subsequently organized by leading research centers in Barcelona together with B•DEBATE, a joint initiative of BIOCAT and “La Caixa” Foundation. Salvador Aznar-Benitah, Eduard Batlle, and Raúl Méndez from the Institute for Research in Biomedicine in Barcelona selected the 2016 BCEC panel of speakers. As the title indicates, this year's conference expanded the epigenetic focus to include additional cancer-relevant topics, such as tumor heterogeneity and RNA regulation. Methods to develop therapeutic approaches on the basis of novel insights have been discussed in great detail. The conference has attracted 217 participants from 11 countries