La señalización purinérgica como modulador de la neurogénesis posnatal y adulta

La neurogénesis se define como la generación de nuevas neuronas funcionales a partir de las células madre neurales (NSCs). Éstas tienen, teóricamente, la capacidad de generar todos los tipos de células gliales y neuronales del sistema nervioso y de autorrenovarse en un número indefinido de veces sin perder su fenotipo y capacidad de diferenciarse (Gage, 2000,Weissman et al., 2001). Al final del desarrollo embrionario la mayoría de las neuronas del Sistema Nervioso Central (SNC) están formadas. No obstante el proceso de neurogénesis continúa tanto en la etapa posnatal como adulta. Numerosos grupos de investigación han tratado de identificar cuál es el comportamiento delas NSCs a lo largo de su progresión de linaje para dar lugar a las distintas poblaciones neurales así como elucidar cuáles son los mecanismos moleculares que controlan el proceso de la neurogénesis. Sin embargo, aún existen numerosos interrogantes por resolver. Entre ellos destacan: i) ¿Hasta qué punto las NSCs son dependientes de programas intrínsecos y/o de las señales provenientes de su nicho neurogénico a la hora de generar sus correspondientes linajes neurales? ii) ¿es una NSC capaz de dar lugar a los tres linajes neurales (astrocitos, oligodendrocitos y neuronas)? o ¿por el contrario presenta restricciones en la elección de su destino celular? iii) ¿Cuáles son las vías de señalización implicadas en la regulación del proceso de neurogénesis? En el presente trabajo pretendemos dar respuesta a algunas de estas cuestiones y analizar la implicación de la señalización mediada por nucleótidos en el comportamiento de las NSCs de dos nichos neurogénicos: el cerebelo posnatal y la zona subependimaria (SEZ) adulta..

Genomic Mapping of Epidermal Growth Factor Receptor and Mesenchymal–Epithelial Transition-Up-Regulated Tumors Identifies Novel Therapeutic Opportunities

Background: The identification of proteins in the cellular membrane of the tumoral cell is a key to the design of therapeutic agents. Recently, the bi-specific antibody amivantamab, targeting the oncogenic membrane proteins EGFR and MET, received regulatory approval for the treatment of adult patients with locally advanced or metastatic NSCLC. Methods: The authors interrogated several publicly available genomic datasets to evaluate the expression of both receptors and PD-L1 in most of the solid and hematologic malignancies and focused on prostate adenocarcinoma (PRAD) and pancreatic adenocarcinoma (PAAD). Results: In PAAD, EGFR highly correlated with PD-L1 and MET, and MET showed a moderate correlation with PD-L1, while in PRAD, EGFR, MET and PD-L1 showed a strong correlation. In addition, in tumors treated with immune checkpoint inhibitors, including anti-PD(L)1 and anti-CTLA4, a high expression of EGFR and MET predicted detrimental survival. When exploring the relationship of immune populations with these receptors, the authors observed that in PAAD and PRAD, EGFR moderately correlated with CD8+ T cells. Furthermore, EGFR and MET correlated with neutrophils in PRAD. Conclusions: The authors identified tumor types where EGFR and MET were highly expressed and correlated with a high expression of PD-L1, opening the door for the future combination of bi-specific EGFR/MET antibodies with anti-PD(L)1 inhibitors

Prostaglandin E2 Impairs P2Y2/P2Y4 Receptor Signaling in Cerebellar Astrocytes via EP3 Receptors

Prostaglandin E2 (PGE2) is an important bioactive lipid that accumulates after tissue damage or inflammation due to the rapid expression of cyclooxygenase 2. PGE2 activates specific G-protein coupled EP receptors and it mediates pro- or anti-inflammatory actions depending on the cell-context. Nucleotides can also be released in these situations and they even contribute to PGE2 production. We previously described the selective impairment of P2Y nucleotide signaling by PGE2 in macrophages and fibroblasts, an effect independent of prostaglandin receptors but that involved protein kinase C (PKC) and protein kinase D (PKD) activation. Considering that macrophages and fibroblasts influence inflammatory responses and tissue remodeling, a similar mechanism involving P2Y signaling could occur in astrocytes in response to neuroinflammation and brain repair. We analyzed here the modulation of cellular responses involving P2Y2/P2Y4 receptors by PGE2 in rat cerebellar astrocytes. We demonstrate that PGE2 inhibits intracellular calcium responses elicited by UTP in individual cells and that inhibiting this P2Y signaling impairs the astrocyte migration elicited by this nucleotide. Activation of EP3 receptors by PGE2 not only impairs the calcium responses but also, the extracellular regulated kinases (ERK) and Akt phosphorylation induced by UTP. However, PGE2 requires epidermal growth factor receptor (EGFR) transactivation in order to dampen P2Y signaling. In addition, these effects of PGE2 also occur in a pro-inflammatory context, as evident in astrocytes stimulated with bacterial lipopolysaccharide (LPS). While we continue to investigate the intracellular mechanisms responsible for the inhibition of UTP responses, the involvement of novel PKC and PKD in cerebellar astrocytes cannot be excluded, kinases that could promote the internalization of P2Y receptors in fibroblasts

Genomic Mapping of Epidermal Growth Factor Receptor and Mesenchymal–Epithelial Transition-Up-Regulated Tumors Identifies Novel Therapeutic Opportunities

The identification of proteins in the cellular membrane of the tumoral cell is a key to the design and guidance of therapeutic agents. Recently, a bi-specific antibody termed amivantamab, targeting epidermal growth factor receptor (EGFR) and mesenchymal–epithelial transition factor (MET), which are oncogenic membrane proteins, received regulatory approval for the treatment of adult patients with locally advanced or metastatic non-small cell lung cancer (NSCLC). In this study, the authors explored tumor types with high levels of expression of EGFR and MET and focused on prostate adenocarcinoma and pancreatic adenocarcinoma, where anti-PD(L)1 agents alone have not shown relevant signs of activity. In addition, the authors confirmed that high expression of either receptor is associated with poor response to anti-PD(L)1 therapy, independently of the expression of PD-L1, suggesting that blocking these receptors with bi-specific EGFR/MET antibodies could augment the efficacy of anti-PD(L)1 inhibitorsBackground: The identification of proteins in the cellular membrane of the tumoral cell is a key to the design of therapeutic agents. Recently, the bi-specific antibody amivantamab, targeting the oncogenic membrane proteins EGFR and MET, received regulatory approval for the treatment of adult patients with locally advanced or metastatic NSCLC. Methods: The authors interrogated several publicly available genomic datasets to evaluate the expression of both receptors and PD-L1 in most of the solid and hematologic malignancies and focused on prostate adenocarcinoma (PRAD) and pancreatic adenocarcinoma (PAAD). Results: In PAAD, EGFR highly correlated with PD-L1 and MET, and MET showed a moderate correlation with PD-L1, while in PRAD, EGFR, MET and PD-L1 showed a strong correlation. In addition, in tumors treated with immune checkpoint inhibitors, including anti-PD(L)1 and anti-CTLA4, a high expression of EGFR and MET predicted detrimental survival. When exploring the relationship of immune populations with these receptors, the authors observed that in PAAD and PRAD, EGFR moderately correlated with CD8+ T cells. Furthermore, EGFR and MET correlated with neutrophils in PRAD. Conclusions: The authors identified tumor types where EGFR and MET were highly expressed and correlated with a high expression of PD-L1, opening the door for the future combination of bi-specific EGFR/MET antibodies with anti-PD(L)1 inhibitors.European CommissionInstituto de Salud Carlos III (España)Fundación CRIS contra el cáncerFundación ACEPAIN contra el CáncerDepto. de Bioquímica y Biología MolecularFac. de Ciencias QuímicasTRUEpubDescuento UC

Combinación de cultivo celular de baja densidad, seguimiento unicelular y patch-clamp para monitorizar el comportamiento de células madre neurales cerebelosas murinas postnatales

Low-density cell culture of the postnatal cerebellum, combined with live imaging and single-cell tracking, allows the behavior of postnatal cerebellar neural stem cells (NSCs) and their progeny to be monitored. Cultured cerebellar NSCs maintain their neurogenic nature giving rise, in the same relative proportions that exist in vivo, to the neuronal progeny generated by the three postnatal cerebellar neurogenic niches. This protocol describes the identification of the nature of the progeny through both post-imaging immunocytochemistry and patch-clamp recordings. For complete details on the use and execution of this protocol, please refer to Paniagua-Herranz et al. (2020b).Government of Madrid (PR65/19-22453)Spanish Ministerio de Ciencia, Innovacioín y Universidades MCIU, (PID2019-109155RB-I00, BFU2015-70067REDC).Sección Deptal. de Farmacología y Toxicología (Veterinaria)Fac. de VeterinariaTRUEpu

Live Imaging Reveals Cerebellar Neural Stem Cell Dynamics and the Role of VNUT in Lineage Progression

Little is known about the intrinsic specification of postnatal cerebellar neural stem cells (NSCs) and to what extent they depend on information from their local niche. Here, we have used an adapted cell preparation of isolated postnatal NSCs and live imaging to demonstrate that cerebellar progenitors maintain their neurogenic nature by displaying hallmarks of NSCs. Furthermore, by using this preparation, all the cell types produced postnatally in the cerebellum, in similar relative proportions to those observed in vivo, can be monitored. The fact that neurogenesis occurs in such organized manner in the absence of signals from the local environment, suggests that cerebellar lineage progression is to an important extent governed by cell-intrinsic or pre-programmed events. Finally, we took advantage of the absence of the niche to assay the influence of the vesicular nucleotide transporter inhibition, which dramatically reduced the number of NSCs in vitro by promoting their progression toward neurogenesis.Ministerio de Educación y Ciencia (BFU2015-70067REDC y BFU2014-53654-P)BRADE-CM (S2013/ICE-2958)UCMSantander (PR26/16-18B-3; PR75/18; PR65/19-22453)Fundación Ramón Areces (PR2018/16-02)7.765 JCR (2020) Q1, 5/29 Cell & Tissue Engineering3.207 SJR (2020) Q1, 23/438 BiochemistryNo data IDR 2020UE

El registro continuo de imagen revela la dinámica de las células madre neurales cerebelosas y el papel de VNUT en la progresión del linaje

Little is known about the intrinsic specification of postnatal cerebellar neural stem cells (NSCs) and to what extent they depend on information from their local niche. Here, we have used an adapted cell preparation of isolated postnatal NSCs and live imaging to demonstrate that cerebellar progenitors maintain their neurogenic nature by displaying hallmarks of NSCs. Furthermore, by using this preparation, all the cell types produced postnatally in the cerebellum, in similar relative proportions to those observed in vivo, can be monitored. The fact that neurogenesis occurs in such organized manner in the absence of signals from the local environment, suggests that cerebellar lineage progression is to an important extent governed by cell-intrinsic or pre-programmed events. Finally, we took advantage of the absence of the niche to assay the influence of the vesicular nucleotide transporter inhibition, which dramatically reduced the number of NSCs in vitro by promoting their progression toward neurogenesis.Comunidad de MadridUniversidad Complutense de MadridFundación Ramón ArecesMinisterio de Educación y Cultura (España)Spanish Ministerio de Ciencia, Innovacio´n y UniversidadesSección Deptal. de Farmacología y Toxicología (Veterinaria)Fac. de VeterinariaTRUEpu

Genomic and Immunologic Correlates in Prostate Cancer with High Expression of KLK2

The identification of surfaceome proteins is a main goal in cancer research to design antibody-based therapeutic strategies. T cell engagers based on KLK2, a kallikrein specifically expressed in prostate cancer (PRAD), are currently in early clinical development. Using genomic information from different sources, we evaluated the immune microenvironment and genomic profile of prostate tumors with high expression of KLK2. KLK2 was specifically expressed in PRAD but it was not significant associated with Gleason score. Additionally, KLK2 expression did not associate with the presence of any immune cell population and T cell activating markers. A mild correlation between the high expression of KLK2 and the deletion of TMPRSS2 was identified. KLK2 expression associated with high levels of surface proteins linked with a detrimental response to immune checkpoint inhibitors (ICIs) including CHRNA2, FAM174B, OR51E2, TSPAN1, PTPRN2, and the non-surface protein TRPM4. However, no association of these genes with an outcome in PRAD was observed. Finally, the expression of these genes in PRAD did not associate with an outcome in PRAD and any immune populations. We describe the immunologic microenvironment on PRAD tumors with a high expression of KLK2, including a gene signature linked with an inert immune microenvironment, that predicts the response to ICIs in other tumor types. Strategies targeting KLK2 with T cell engagers or antibody–drug conjugates will define whether T cell mobilization or antigen release and stimulation of immune cell death are sufficient effects to induce clinical activity

Searching for motivational keys to the study of Pharmacology: meaningful learning with Escape Room

En plena era de una transformación digital constante, las estrategias educativas y los nuevos métodos de enseñanza se han diversificado exponencialmente tanto dentro como fuera de las aulas. El objetivo final y global de la implementación de nuevas prácticas educativas en el ámbito universitario es favorecer el proceso de enseñanza–aprendizaje y, por consiguiente, mejorar los resultados académicos. A través de metodologías activas en el ámbito de la educación, como en nuestra propuesta, la interrelación existente entre gamificación, dopamina y motivación es un determinante de la atención. a la atención determinante. Estas metodologías abren nuevos horizontes a la enseñanza y a los métodos de aprendizaje permitiendo una optimización de esfuerzos para alcanzar las competencias y motivando la participación, cohesión y liderazgo de las/os estudiantes (Zhang et al., 2018). En este contexto, queremos acercar a los estudiantes a juegos virtuales en equipos bajo los esquemas de Escape Room, actividad ampliamente difundida en muchos contextos no educativos dentro de los jóvenes. En su adaptación al proceso enseñanza-aprendizaje de la Farmacología, se propone la elaboración de “salas de escape” en las que los estudiantes tienen que superar una serie de pruebas para “escapar” cumpliendo los objetivos requeridos, mediante la resolución de puzles, enigmas, conectando pistas o abriendo candados con clave, todo ello utilizando los conocimientos teórico-prácticos relacionados con contenidos de la asignatura de Farmacología y Farmacia y de Farmacología Clínica y Farmacoterapéutica Veterinaria en una escala de espacio-tiempo limitado. Este método promueve el desarrollo de habilidades mentales para la solución de enigmas y problemas –de índole farmacológico- de manera que las/os estudiantes pongan en juego la creatividad y el pensamiento crítico, hecho que incrementa la motivación de los participantes en las asignaturas de Farmacología cuando los conocimientos teóricos y abstractos ven una aplicabilidad más real que les permite ser capaces de resolver los retos que se les plantean y proporciona un sentimiento de control y autonomía. Además, mediante una estrecha colaboración con los profesores, entre los pasos a seguir, se llevarán a cabo búsquedas de información en páginas web institucionales como el Sistema de Bibliotecas de la Universidad Complutense de Madrid (UCM), lecturas recomendadas y revistas científicas que puedan aportar pistas para la resolución de los problemas, además de reforzar el aprendizaje de la asignatura y la utilización de las fuentes de información disponibles en la Universidad. Este proyecto surge como una forma de amenizar e innovar el aprendizaje de la Farmacología que tradicionalmente se caracteriza por la memorización y asimilación de gran cantidad de conceptos complejos y abstractos. Nuestra comunidad educativa debe sacar provecho de la información disponible en la red y de los nuevos medios relacionados con la tecnología educativa con el fin de mejorar el rendimiento académico de las/os estudiantes de Farmacología Veterinaria. La enseñanza de la Farmacología se presta bien al empleo del aprendizaje cooperativo ya que dicha materia se encuentra en la interfase entre las disciplinas básicas (Anatomía, Fisiología, Biología, Bioquímica...) y las clínicas (Patología, Cirugía...) de las distintas titulaciones en Ciencias de la Salud (Medicina, Veterinaria, Farmacia...), permitiendo así una integración de los conocimientos aprendidos en las diferentes asignaturas que cursan. Una forma de integrar eficazmente los aspectos básicos y clínicos, y facilitar su aprendizaje profundo (cooperativo, interactivo...) es la elaboración de escenarios-problema (“salas de escape”) que permitan, tras el aprendizaje de los conocimientos teóricos, el análisis de las múltiples circunstancias reales que condicionan la elección del mejor tratamiento farmacológico (claves, resolución de acertijos, etc.) en situaciones clínicas relevantes y su enseñanza, de forma que sean los estudiantes quienes realicen el análisis y su autoaprendizaje.In an era of constant digital transformation, educational strategies and new teaching methods have diversified exponentially both inside and outside the classroom have diversified exponentially both inside and outside the classroom. The ultimate and overarching goal of the implementation of new educational practices in the university environment is to favour the teaching-learning process and, consequently, to improve academic results. Through active methodologies in the field of education, as in our proposal, the interrelationship between gamification, dopamine and motivation as a determinant of attention. These methodologies open up new horizons in teaching and learning methods, allowing for the learning methods, allowing an optimisation of efforts to achieve competences and motivating participation, cohesion and leadership of learners (Zhang et al., 2018). In this context, we want to bring learners closer to virtual games in teams under the schemes of Escape Room, an activity widely spread in many non-educational contexts among young people. In its adaptation to the Pharmacology teaching-learning process, we propose the development of "escape rooms" in which the students have to overcome a series of students have to overcome a series of tests in order to "escape" by fulfilling the required objectives, by means of the solving puzzles, enigmas, connecting clues or opening padlocks with a key, all using the theoretical and practical knowledge related to the Theoretical and practical knowledge related to the contents of the subject Pharmacology and Pharmacy and Clinical Pharmacology and Pharmacotherapy in a limited space-time scale. This method promotes the development of mental skills for the solution of enigmas and problems - of a pharmacological nature - in such a way that promotes the development of mental skills in solving pharmacological scenarios and problems so that students bring creativity and critical thinking into play, which increases the motivation of subject participants. Motivation of participants in pharmacology subjects, when theoretical and abstract knowledge is given, a more real-world applicability that allows them an applicability that enables them to be able to solve the challenges they face and provides a sense of control and autonomy. In addition, through close collaboration with the teachers, among the steps to be taken will included tools for information searching on institutional websites such as the Library System of the Complutense University of Madrid (UCM), and recommended readings and scientific journals that can provide clues for the resolution of the problems, as well as reinforcing the learning of the subject and the use of the information sources available at the University. This project arises as a way to liven up and innovate the learning of Pharmacology, which is traditionally characterised by the memorisation and assimilation of a large number of complex and abstract concepts. Our educational community must take advantage of the information available on the Internet and new media related to educational technology in order to improve the academic performance of students of Veterinary Pharmacology. The teaching of Pharmacology lends itself well to the use of cooperative learning as the subject is at the interface between the subject of basic disciplines (Anatomy, Physiology, Biology, Biochemistry...) and clinical disciplines (Pathology, Surgery...). (Pathology, Surgery...) of the different degrees in Health Sciences (Medicine, Veterinary Medicine, Pharmacy...), thus allowing an integration of the knowledge learnt in the different subjects they take. A way of effectively integrating basic and clinical aspects, and facilitating deep learning (cooperative, interactive...). The development of problem-scenarios is the elaboration of "escape rooms" (clues, solving puzzles, etc.) which allow, after learning the theoretical master class, the analysis of the multiple real circumstances that conditions the choice of the best pharmacological treatment (keys, solving the pharmacological treatment) in relevant clinical situations, so the students are who carry out the analysis and self-learning.Sección Deptal. de Farmacología y Toxicología (Veterinaria)Fac. de VeterinariaFALSEUniversidad Complutense de Madridsubmitte