Glia and neurons from human iPSCs to address the pathology of Alzheimer´s disease

Alzheimer's disease (AD) is characterized by presenting a complex pathology, not fully resolved yet. This fact, together with the lack of reliable models, has impeded the development of effective therapies. Recently, several studies have shown that functional glial cell defects have a key role in the pathology of AD. However, this glial dysfunction, currently, cannot be correctly modeled using the available animal models, so we hypothesized that cells derived from Alzheimer's patients can serve as a better platform for studying the disease. In this sense, human pluripotent stem cells (hPSC) allow the generation of different types of neural cells, which can be used for disease modeling, identification of new targets and drugs development. Methods: We have a collection of hiPSCs derived from patients with sporadic forms of AD stratified based on APOE genotype. We have differentiated these cells towards neural cells and mature them to neurons or astrocytes using a serum-free approach, to assess intrinsic differences between those derived from AD patients or healthy controls. Results: We have implemented a serum-free approach and generated neural precursors and astrocytes from all the lines tested. We observe differences at the phenotypic level and a reduced capacity to differentiate towards neural lineage in those lines derived from APOE4 carriers. Conclusions: Our preliminary data suggest intrinsic differences in the neural differentiation capacity between cell lines derived from APOE4 or APOE3 carrier subjects. Further experiments would contribute to elucidate novel pathogenic pathways associated with neurodegeneration and susceptible of therapeutic modulation, likely contributing to the development of new effective drugs against AD.This study was supported by Instituto de Salud Carlos III (ISCiii) of Spain, co-financed by FEDER funds from European Union, through grants PI21/00915 (to AG) and PI21/00914 (to JV); by Junta de Andalucia through Consejería de Economía y Conocimiento grants UMA20-FEDERJA-048 (to JAGL), PY18-RT-2233 (to AG) and US-1262734 (to JV) co-financed by Programa Operativo FEDER 2014-2020, and Programa Operativo de Empleo Juvenil SNGJ4-11 to LCP. Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Neurons and astrocytes derived from human iPSCs to model Alzheimer´s disease

Alzheimer's disease (AD) is characterized by presenting a complex pathology, not fully resolved yet. This fact, together with the lack of reliable models, has impeded the development of effective therapies. Recently, several studies have shown that functional glial cell defects have a key role in the pathology of AD. However, this glial dysfunction, currently, cannot be correctly modeled using the available animal models, so we hypothesized that cells derived from Alzheimer's patients can serve as a better platform for studying the disease. In this sense, human pluripotent stem cells (hPSC) allow the generation of different types of neural cells, which can be used for disease modeling, identification of new targets and drugs development. Methods: We have a collection of hiPSCs derived from patients with sporadic forms of AD stratified based on APOE genotype. We have differentiated these cells towards neural cells and mature them to neurons or astrocytes using a serum-free approach, to assess intrinsic differences between those derived from AD patients or healthy controls. Results: We have implemented a serum-free approach and generated neural precursors and astrocytes from all the lines tested. We observe differences at the phenotypic level and a reduced capacity to differentiate towards neural lineage in those lines derived from APOE4 carriers. Conclusions: Our preliminary data suggest intrinsic differences in the neural differentiation capacity between cell lines derived from APOE4 or APOE3 carrier subjects. Further experiments would contribute to elucidate novel pathogenic pathways associated with neurodegeneration and susceptible of therapeutic modulation, likely contributing to the development of new effective drugs against AD.This study was supported by Instituto de Salud Carlos III (ISCiii) of Spain, co-financed by FEDER funds from European Union, through grants PI21/00915 (to AG) and PI21/00914 (to JV); by Junta de Andalucia through Consejería de Economía y Conocimiento grants PY18-RT-2233 (to AG) and US-1262734 (to JV) co-financed by Programa Operativo FEDER 2014-2020, Consejeria de Salud grant PI-0276-2018 (to JAGL) and Programa Operativo de Empleo Juvenil SNGJ4-11 to LCP. Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Co-cultures between neurons and astrocytes to address Alzheimer´s disease pathology.

Background: Alzheimer's disease (AD) is characterized by presenting a complex pathology, not fully resolved yet. This fact, together with the lack of reliable models, has impeded the development of effective therapies. Recently, several studies have shown that functional glial cell defects have a key role in the pathology of AD. However, this glial dysfunction, currently, cannot be correctly modeled using the available animal models, so we hypothesized that cells derived from Alzheimer's patients can serve as a better platform for studying the disease. In this sense, human pluripotent stem cells (hPSC) allow the generation of different types of neural cells, which can be used for disease modeling, identification of new targets and drugs development. Methods: We have a collection of hiPSCs derived from patients with sporadic forms of AD stratified based on APOE genotype. We have differentiated these cells towards neural cells and mature them to neurons or astrocytes using a serum-free approach, to assess intrinsic differences between those derived from AD patients or healthy controls. Results: We have implemented a serum-free approach and generated neural precursors and astrocytes from all the lines tested. We observe differences at the phenotypic level and a reduced capacity to differentiate towards neural lineage in those lines derived from APOE4 carriers. Conclusions: Our preliminary data suggest intrinsic differences in the neural differentiation capacity between cell lines derived from APOE4 or APOE3 carrier subjects. Further experiments would contribute to elucidate novel pathogenic pathways associated with neurodegeneration and susceptible of therapeutic modulation, likely contributing to the development of new effective drugs against AD.This study was supported by ISCiii (Spain), co-financed by FEDER funds, through grants PI21/00915 (AG) and PI21/00914 (JV); by Junta de Andalucia through Consejería de Economía and Conocimiento grants UMA20-FEDERJA-048 (JAGL), PY18-RT-2233 (to AG) and US-1262734 (JV), co-financed by Programa Operativo FEDER 2014-2020, and SNGJ4-11 (LCP). Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Mitochondrial ultrastructural defects in reactive astrocytes of Alzheimer's mice hippocampus.

Alzheimer's disease (AD) is a complex neurodegenerative condition that causes progressive memory loss and dementia. In AD brains astrocyte become reactive potentially contributing to cognitive decline. Astrocyte reactivity is a highly complex phenomenon with remarkable morphologic and molecular phenotype changes, and the role of astrocytes in the development of AD is still unknown. Astrocytes are the prevalent glial cells in the brain and have a large number of functions aimed at maintaining brain homeostasis including regulation of brain energy metabolism, maintenance of the blood-brain barrier, ion homeostasis, synaptic activity and plasticity, among many other functions. Any disruption regarding the normal roles of astrocytes can result in morphological and functional changes that ensue in pathological consequences. Mitochondrial dysfunction is an early event in the pathogenesis of AD, although most studies have focused on neurons and little is known about the functional characteristics and the dynamics of astrocyte mitochondria. We had performed an ultrastructural analysis using transmission electron microscopy in the hippocampus of amyloidogenic (APP/PS1) and tauopathy (P301S) mice. Our results show structural alterations in mitochondria that include double membrane rupture, cristae loss, and fragmentation together with a loss of their circularity. Since mitochondrial morphology is directly related to mitochondrial fusion/fission processes, the ultrastructural changes observed in astrocyte mitochondria in these amyloidogenic and tauopathy models suggest dynamic abnormalities in these organelles that may lead to deficits in astroglial function compromising their capability to maintain brain homeostasis and support neuronal energy metabolism and survival. A better understanding of cell type-specific mitochondrial dysfunction as a pathological feature of AD might hold great potential for the exploration of novel molecular targets for therapeutic development.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Articulación agroecológica: estrategias para la reconversión a escala territorial

Entre los principales argumentos que pretenden descalificar una agricultura sustentable están sus bajos rendimientos y elevados costos. Además, se argumenta una limitada aplicabilidad (solo a escala de pequeñas fincas). Esta polémica se halla presente en la Cuba actual, urgida de soluciones a corto plazo para la alimentación de toda su población. Por ello, se necesita disponer de estrategias y modelos de desarrollo que respondan a los requerimientos actuales, asumiendo el reto de diseñar una agricultura sostenible sobre bases agroecológicas, productiva y eficiente, aplicable a diferentes condiciones y escalas productivas. Este fue reto del proyecto “Articulación agroecológica; diseño de alternativas sostenibles para la seguridad alimentaria local” promovido por la Asociación Cubana de Técnicos Agrícolas y Forestales (ACTAF) y la ONG Holandesa HIVOS. Esta acción se viene ejecutando desde hace 4 años (2011-2015) en 5 municipios cubanos, con apoyo financiero de la Unión Europea, de la Colaboración Suiza para el Desarrollo (COSUDE) y del Ministerio de la Agricultura de Cuba (MINAG). Se partió de la hipótesis de que el desarrollo agrario sostenible sobre bases agroecológicas tiene que diseñarse y conducirse localmente (escala municipal). Para ello, se planteó como objetivo, “contribuir al incremento y la sostenibilidad de la seguridad alimentaria cubana, facilitando la adopción de alternativas de articulación agroecológica en las estrategias de desarrollo agrario municipal”. Para el logro de este objetivo se planteó la necesidad de crear y fortalecer capacidades locales de desarrollo agrario, a partir de instituciones existentes y en coherencia con los programas oficiales de la agricultura local y los intereses y necesidades sentidas de los agricultores. Se trabajó desde un Equipo de Proyecto apoyado por un Equipo Técnico conformado por especialistas de diferentes entidades científicas y docentes nacionales. A nivel local se conformaron Equipos de Articulación Local (EAL) con integrantes designados por cada municipio. Estos EAL constituyeron el principal ente implementador del proyecto y se concibieron como el germen inicial de lo que debía ser el Equipo Municipal de Desarrollo Agrario de cada territorio. Alrededor de estos EAL se articularon las principales instituciones locales vinculadas a la agricultura y se seleccionó una cooperativa para demostrar la pertinencia de la reconversión agroecológica a escala superior a la pequeña finca. Se pudo comprobar que lograr el involucramiento de decisores importantes de la localidad es del todo imprescindible para un proceso de reconversión agroecológica a escala territorial, pues este debe ser visto como contribuyente al logro de los objetivos de las instituciones locales para la seguridad alimentaria de la población y minimizar la dependencia externa al territorio para el desarrollo de toda la cadena productiva. Disponer de servicios técnicos locales que puedan contribuir con insumos alternativos, asesoría y capacitación, aporta a la reconversión capacidad de contextualización de conocimientos y tradiciones campesinas y establece puentes con instituciones científicas especializadas para su complementación con los avances de la ciencia y la técnica.Eje: B2 Paisajes, territorios y agroecología (Relatos de experiencias)Facultad de Ciencias Agrarias y Forestale

Astroglial reactivity in response to b-amyloidosis is associated with mithocondrial pathology in the hippocampus of Alzheimer's transgenic mice

Aims: Alzheimer’s diseaase (A)D is associated with early energy hypometabolism, synaptic and mitochondrial dysfunction, oxidative stress, inflammation, abnormal proteostasis and progressive neurodegeneration. During the pathogenic process, amyloid-beta and phospho-tau pathologies have a detrimental effect on neurons and glial cells, affecting neuronal stability, and compromising ATP production and energy metabolism. Though mitochondrial dysfunction is thought to be an early event in the pathogenesis of this AD, the vast majority of studies have focused on neurons, and little is known about the functional characteristics and dynamics of mitochondria in astrocytes. Here, we aim to analyze mitochondrial subcellular features of reactive astrocytes in APP/PS1 mice hippocampus in order to a better understanding of this pathology. Methods: Mitochondrial features were observed by transmission electron microscopy and immunogold labeling. Image analysis was performed to assess morphological changes. Results: Our results show mitochondrial structural alterations including mitochondrial cristae loss, broken double membrane structure and fragmentation. In addition, an increase in both number and size of mitochondria in this transgenic model compared to age-matched WT mice, was found. Conclusions: Since mitochondrial morphology is directly related to mitochondrial fusion/fission, the ultrastructural pathology of astrocytic mitochondria in this amyloidogenic model suggests dynamics abnormalities in these organelles that might lead to astroglial functional deficits compromising neuronal survival. Supported by ISCiii grants PI21-0915 (AG) and PI21-00914 (JV) co-financed by FEDER funds from European Union, by Junta de Andalucia grants P18-RT-2233 (AG) and US-1262734 (JV) co-financed by Programa Operativo FEDER 2014-2020, and by grant PPIT.UMA.B1-2021_32 (LTE).Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech