Person:
Rosca, Andreea

First Name

Andreea

Last Name

Rosca

Institution

ISCIII

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ISCIII::Unidad Funcional de Investigación de Enfermedades Crónicas (UFIEC)

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Now showing 1 - 10 of 11

Impact of environmental neurotoxic: current methods and usefulness of human stem cells.
(Elsevier, 2020-12) Rosca, Andreea; Coronel Lopez, Raquel; Moreno, Miryam; Gonzalez-Martin-Niño, Rosa Maria; Oniga, Andreea; Martin, Alberto; López-Alonso, Victoria; González-Caballero, MCarmen; Liste-Noya, Isabel; Ministerio de Ciencia e Innovación (España); Ministerio de Economía y Competitividad (España); Comunidad de Madrid (España)
The development of central nervous system is a highly coordinated and complex process. Any alteration of this process can lead to disturbances in the structure and function of the brain, which can cause deficits in neurological development, resulting in neurodevelopmental disorders, including, for example, autism or attention-deficit hyperactivity disorder. Exposure to certain chemicals during the fetal period and childhood is known to cause developmental neurotoxicity and has serious consequences that persist into adult life. For regulatory purposes, determination of the potential for developmental neurotoxicity is performed according the OECD Guideline 426, in which the test substance is administered to animals during gestation and lactation. However, these animal models are expensive, long-time consuming and may not reflect the physiology in humans; that makes it an unsustainable model to test the large amount of existing chemical products, hence alternative models to the use of animals are needed. One of the most promising methods is based on the use of stem cell technology. Stem cells are undifferentiated cells with the ability to self-renew and differentiate into more specialized cell types. Because of these properties, these cells have gained increased attention as possible therapeutic agents or as disease models. Here, we provide an overview of the current models both animal and cellular, available to study developmental neurotoxicity and review in more detail the usefulness of human stem cells, their properties and how they are becoming an alternative to evaluate and study the mechanisms of action of different environmental toxicants.
Background levels and brain organoid impact of RF field exposure in a healthcare environment
(Frontiers Media, 2024-03) Hernández, José A.; Rosca, Andreea; Suárez, Samuel; Coronel Lopez, Raquel; Suarez, Oscar J.; Peran-Ramos, Paula; Marina-Boillos, Pablo; Rabassa, Luis E.; Mateos-Martínez, Patricia; Liste-Noya, Isabel; López-Alonso, Victoria; Torres-Ruiz, Mónica; Febles, Victor M.; Ramos-Gonzalez, Maria Victoria; Instituto de Salud Carlos III; Unión Europea. Fondo Europeo de Desarrollo Regional (FEDER/ERDF); Agencia Estatal de Investigación (España)
Introduction: This study is an introduction to the empirical and impact evaluation of radiofrequency electromagnetic field (RF-EMF) radiation exposure in a healthcare environment, focusing on an indoor microenvironment. It explores the expression of various genes associated with cellular responses, cell proliferation, senescence, and apoptotic cell death. The assessment analyzes current personal mobile communications (2G-5G FR1), providing a clear understanding of RF-EMF exposure and compliance with regulatory limits. Methods: The signals from different wireless communication systems at Hospital Universitario de Canarias (HUC) in Tenerife, Canary Islands, Spain, were examined in 11 locations. Four measurement campaigns were performed with frequencyselective exposimeters (PEMs) and an EME Spy 200 MVG, and experimental electric field values were compared as a long-term exposition. The frequency with the highest contribution (2.174 V/m) observed (1840 MHz) in UMTS was selected for biological effects evaluation. Results: The study focuses on four locations with the highest exposure to communication systems (downlinks), analyzing the results to verify compliance with regulations that ensure the safety of patients, the general public, and healthcare workers. LTE B20 (DL), GSM+UMTS 900 (DL), GSM 1800 (DL), UMTS 2100 (DL), and LTE B7 (DL) exhibited relatively higher E/m values throughout the campaigns, and these values consistently remained below the ICNIRP reference levels, signifying a consistently low level of exposure. In addition, this work presents the biological effects on neural stem cells (NSCs) using 3D brain organoids (BOs) exposed to RF signals in a validated and commercial experimental setting: the Gigahertz Transverse Electromagnetic cell (GTEM). The GTEM allows for the creation of homogeneous field electromagnetic fields in a small, enclosed setting and guarantees exposure conditions in a wide range of frequencies. BOs are an in vitro 3D cell-culture technology that reproduces the cellular composition and structure of the developing brain. Analyzing the expression of several genes associated with cellular responses, cell proliferation, senescence, and apoptotic cell death,wefound that exposure of BOs at 1840MHzdid not affectmRNAexpression in brain genes related to apoptosis or senescence. However, a decrease in gene expression for cell proliferation and cell activity markers was observed during the differentiation stage of BOs. Discussion: The discussion emphasizes the coexistence and evolution of various heterogeneous networks and services throughout the four measurement campaigns. Across all measured results, the levels of the obtained E-field were consistently well below the exposure limits set by internationally accepted standards and guidelines. These obtained values have been established in order to consider their potential effects on cell proliferation and cell activity, especially in differentiating biological organisms. Consequently, the results obtained and the methodology presented could serve as a foundational framework for establishing the basis of RF-EMF assessment in future heterogeneous 5G developments, particularly in the millimeter wave (mmWave) frequency range, where the forecast is for massive high-node density networks.
RNA-SEQ analysis reveals the implication of Amyloid Precursor Protein (APP) in cell fate specification of human neural Stem Cells by several Signaling Pathways. IBRO Neuroscience Reports
(Elsevier, 2023) Coronel Lopez, Raquel; Rosca, Andreea; González-Sastre, Rosa; Mateos-Martínez, Patricia; Gallego, Marta Ines; Liste-Noya, Isabel; López-Alonso, Victoria
Numerous studies have focused on the pathophysiological role of amyloid precursor protein (APP) in Alzheimer's disease (AD), being the aggregation of β-amyloid (Aβ) peptide a central event. However, many authors consider that alterations in physiological functions of APP are likely to play a key role in AD pathogenesis. APP is a glycoprotein expressed ubiquitously in a wide variety of tissues, being especially abundant in the brain. The APP expression is detected at early stages of nervous system development, as well as in adult brain, suggesting that this protein plays a key and important role at various stages of life. Previous studies in our laboratory revealed that APP playing an important role on differentiation of human neural stem cells (hNSCs), favoring glial differentiation (gliogenesis), and preventing the differentiation of them towards neuronal phenotype (neurogenesis). In that case, we proposed to the APP/AICD/GSK-3β system as a possible molecular mechanism involved in the observed effects. However, given the multifunctionality of APP, we think that other molecular mechanisms could also be implicated. In the present study, we have evaluated the effects of APP overexpression in hNSCs at a global level by a transcriptomic analysis using the massive RNA sequencing (RNA-seq) technology. Specifically, we have focused on differentially expressed genes (DEGs) that are related to neuronal and glial differentiation processes, as well as on groups of DEGs associated with different signaling pathways, to find a possible interaction between them and APP. Our data indicate a differential expression in genes related to gliogenesis and neurogenesis processes, as well as in the pathways of Notch signaling, Wnt signaling, PI3K-AKT signaling, and JAK-STAT signaling, among other. The knowledge of physiological functions of APP, as well as the possible signaling pathways that could be implicated, are essential to advance the understanding of the pathogenesis of AD.
SINGLE-CELL sequencing workflow to study cellular composition and cell type specific expression profiles of human Cerebral Organoids
(Elsevier, 2023) González-Sastre, Rosa; Coronel Lopez, Raquel; Mateos-Martínez, Patricia; Jimenez Sancho, Maria Pilar; Rosca, Andreea; Maeso, Laura; Martín Benito, Sabela; Zaballos, Ángel; Liste-Noya, Isabel; López-Alonso, Victoria
Human cerebral organoid culture is a technology with immense potential in the areas of developmental neurobiology and neurodegeneration for example to study cell types, mechanisms involved, to discover of new biomarkers, to propose specific therapeutic strategies or to study the effects of compound-induced toxicity. Single-cell RNA sequencing (scRNA-seq) is a promising technology that will help to define the identity of the cerebral organoids and to understand cellular composition and cell type specific expression profiles. Standardization of workflows to do the scRNA-seq analysis is an important means to improve the use of this technology. We present the workflow and results of the scRNA-seq performed for cerebral organoids generated from the AND-2 cell line of human embryonic stem cells (hESCs). Dissociated cerebral organoid samples were loaded on the 10X Chromium and single cell libraries were prepared according to 10X Genomics standard procedures and sequenced on the Novaseq sequencer (Illumina).The data were checked and aligned to the GRCh38 human reference genome with CellRanger v6.0.2 and analyzed with Seurat v4.0. After quality filtering and data normalization with the SCTransform function, we performed Principal component analysis (PCA) using the highly variable genes, built a Shared Nearest Neighbor (SNN) graph using the Louvain method. To visualize data, Uniform Manifold Approximation and Projection (UMAP) dimensional reduction was performed. The identities of the cell clusters were assigned using the expression of genes specific of each cell type. We annotate in the AND2 cerebral organoids clusters for intermediate progenitor cells, astrocytes, oligodendrocyte precursor cells, excitatory neurons, inhibitory neurons, and mesodermal cells. We find also some cells in these organoids with expression of endothelial and microglial gene markers. Enrichment analysis of the highly variable differentially expressed genes (DEGs) was utilized to characterize the assigned cell types with Gene Ontology (GO), PanglaoDB and Cellmarker databases.
Advances in Alzheimer’s Disease Research: Human Cerebral Organoids
(Biomedgrid, 2023) Mateos-Martínez, Patricia; González-Sastre, Rosa; Coronel Lopez, Raquel; Rosca, Andreea; Martín Benito, Sabela; Bernabeu-Zornoza, Adela; López-Alonso, Victoria; Liste-Noya, Isabel; Ministerio de Ciencia e Innovación (España); Instituto de Salud Carlos III; Ministerio de Ciencia, Innovación y Universidades (España)
Alzheimer’s disease (AD) is the main neurodegenerative disorder in old age, causing memory impairment and dependency. The histopathology of AD is characterized by the presence of amyloid plaques and neurofibrillary tangles formed by Aβ peptide and hyperphosphorylated Tau, respectively. There is still no cure or effective treatment for AD. This could be due, in part, to the lack of suitable research models since animal models do not recapitulate the full physiological complexity of the human brain. With the development of induced pluripotent stem cells (iPSCs), these limitations could be overcome. Even so, the bi-dimensional (2D) culture models still do not allow to recapitulate all types of brain cells and do not show a three-dimensional (3D) arrangement. Since obtaining 3D cultures called organoids, a new opportunity arises to overcome the limitations of previous models. Human Cerebral Organoids (hCOs) represent a pioneering model, in which part of the complexity of the human brain is present. For this reason, they are fast becoming a very remarkable model for the study of the evolution of the molecular and cellular pathology of AD. This review provides a brief overview of AD research, focusing on the most recent advances achieved through the development of stem cell and cerebral organoid technology
Studying the cellular and molecular evolution of familiar Alzheimer'sdisease using human Cerebral Organoids
(Elsevier, 2023) Mateos-Martínez, Patricia; González-Sastre, Rosa; Coronel Lopez, Raquel; Rosca, Andreea; Maeso, Laura; Martín Benito, Sabela; Luque, Daniel; Terrón, Maria Carmen; Alonso, Victoria López; Liste, Isabel
The leading cause of dementia in the elderly is Alzheimer's disease (AD), and its increase is expected in the coming years. The histopathological hallmarks of AD are associated with the presence in brain of neurofibrillary tangles, due to the increase in hyperphosphorylated Tau protein, as well as amyloid plaques, due to the increase in amyloid peptide. There is no cure for AD and the treatments effective in slowing neurodegeneration. This lack of cure/treatments may be due to the lack of good study models. Until now, in vivo or in vitro monolayer cellular models have been used that do not allow recapitulating the complexity of the human brain, as well as the histopathology of AD in early stages of its development. For all these reasons, in this work we consider using the technology of three-dimensional cultures: human cerebral organoids (hCOs). In this work, using the protocol developed in our laboratory, we present the generation and characterisation of hCOs with mutations associated with familial AD (fAD) as compared with control hCOs. For this purpose, we used human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs) as controls. As fAD models, we used hiPSCs with mutations in PSEN1 and APP duplication. Using immunohistochemistry and RT-qPCR we have analysed these hCOs for markers of neural precursors, brain cell types and synaptic markers, as well as the progression of AD phenotype (amyloid plaques). We have also performed electron microscopy studies to observe differences in the ultrastructure of the hCOs. We present the differences found that show the hCOs generated from hiPSCs with AD variants are experimental in vitro models that will allow further study of pathology.
Neurodevelopmental effects of Cypermethrin in human Neural Stem Cells
(Elsevier, 2023) Rosca, Andreea; Coronel Lopez, Raquel; González-Sastre, Rosa; Mateos-Martínez, Patricia; López-Alonso, Victoria; González-Caballero, MCarmen; Liste-Noya, Isabel
The lack of adequate models for the study of developmental neurotoxicity has made difficult to correctly assess the risk and effects of toxic exposure. Despite the large body of results on animals, these studies are costly, time consuming and the results are not always reliable to assess the impact of chemical compounds on the developing human brain because animal models do not perfectly reflect human physiology. In vitro systems are becoming a promising tool to assess the toxicological effects of chemical compounds on developmental neurotoxicity with promising results. Currently, stem cells are becoming a useful model to study this type of toxicity. Stem cells are undifferentiated cells with the potential to differentiate into more specialized cell types. They are present during brain development and into adult life, making them a more appropriate model for mimicking key events that take place during embryonic brain development. Cypermethrin (CYP) is one of the most widely used and highly effective synthetic pyrethroids. CYP can enter the body mainly through skin contact but also through inhalation or ingestion of food or water. The main mechanism of action of pyrethroids is the interaction with Na channels and the induction of prolonged depolarization in neurons. In this study, the human neural stem cell line hNS1 was used to evaluate the effects of CYP on early developmental stages. hNS1 cells were exposed to different concentrations of the pesticide and cell death, proliferation and cell fate specification were analyzed under differentiation conditions by immunocytochemistry and RT-qPCR. The results showed that this compound induces apoptotic cell death at the highest doses tested and a decrease in cells in the cell cycle. Besides, CYP causes a decrease in both neurogenesis and gliogenesis in hNS1 cells. In conclusion, CYP has toxic effects on hNS1 cells and should be further studied.
Highly efficient generation of human Cerebral Organoids bypassing embryoid body stage
(Elsevier, 2023) González-Sastre, Rosa; Coronel Lopez, Raquel; Bernabeu-Zornoza, Adela; Rosca, Andreea; Mateos-Martínez, Patricia; Maeso, Laura; Martín Benito, Sabela; López-Alonso, Victoria; Liste, Isabel
Human cerebral organoids (hCOs) are a promising in vitro model that may overcome some of the limitations that currently exist when studying human brain development and disease. Since Lancaster et al. first generated hCOs, efforts have been made to better recapitulate the physiology of the human brain and improve the efficiency and reproducibility of protocols. Different groups employed dual-SMAD inhibition (double inhibition of the transforming growth factor-β (TGF-β) and bone morphogenetic proteins (BMPs) pathways) to achieve rapid neural induction. The method developed here, outlines the generation of homogeneous organoids by rapid neuroepithelial induction, avoiding the Embryoid Body (EB) stage. The efficiency of this protocol to form neuroepithelial structures and subsequently organoids is almost 100% due to the use of dual-SMAD inhibition in combination with CHIR99021 (a GSK3β inhibitor/Wnt activator) at the neural induction stage. This is a simple and reproducible protocol as we do not need to use Matrigel or bioreactors which standardizes the methodology. It is also a robust protocol as we have successfully performed it on human embryonic stem cells (hESCs) and human induced pluripotent stem cells (iPSCs). We performed IHC and Q-RT-PCR assays for cell cycle, neural precursors, neuronal and synaptic vesicle markers. The generated hCOs are highly homogeneous and show ventricular zones (VZs) with radial glia in the center that differentiate to give rise to neurons located around these VZs. These neurons acquire a mature state and are able to form synapses. In addition, we also performed IHC and Q-RT-PCR assays for different markers of oligodendrocytes, astrocytes, microglial cells and vasculature obtaining promising results. We believe that this protocol will be a breakthrough in the generation of organoids for use as a model for the study of neurodevelopmental and neurological diseases, as well as for drug testing.
Physiological effects of amyloid precursor protein and its derivatives on neural stem cell biology and signaling pathways involved
(Medknow Publications, 2019-10) Coronel Lopez, Raquel; Palmer, Charlotte; Bernabeu-Zornoza, Adela; Monteagudo, Maria; Rosca, Andreea; Zambrano, Alberto; Liste-Noya, Isabel; Instituto de Salud Carlos III; Ministerio de Economía y Competitividad (España); Comunidad de Madrid (España)
The pathological implication of amyloid precursor protein (APP) in Alzheimer's disease has been widely documented due to its involvement in the generation of amyloid-β peptide. However, the physiological functions of APP are still poorly understood. APP is considered a multimodal protein due to its role in a wide variety of processes, both in the embryo and in the adult brain. Specifically, APP seems to play a key role in the proliferation, differentiation and maturation of neural stem cells. In addition, APP can be processed through two canonical processing pathways, generating different functionally active fragments: soluble APP-α, soluble APP-β, amyloid-β peptide and the APP intracellular C-terminal domain. These fragments also appear to modulate various functions in neural stem cells, including the processes of proliferation, neurogenesis, gliogenesis or cell death. However, the molecular mechanisms involved in these effects are still unclear. In this review, we summarize the physiological functions of APP and its main proteolytic derivatives in neural stem cells, as well as the possible signaling pathways that could be implicated in these effects. The knowledge of these functions and signaling pathways involved in the onset or during the development of Alzheimer's disease is essential to advance the understanding of the pathogenesis of Alzheimer's disease, and in the search for potential therapeutic targets.
Amyloid Precursor Protein (APP) Regulates Gliogenesis and Neurogenesis of Human Neural Stem Cells by Several Signaling Pathways
(Multidisciplinary Digital Publishing Institute (MDPI), 2023-08-19) Coronel Lopez, Raquel; Bernabeu-Zornoza, Adela; Palmer, Charlotte; González-Sastre, Rosa; Rosca, Andreea; Mateos-Martínez, Patricia; López-Alonso, Victoria; Liste-Noya, Isabel; Ministerio de Ciencia e Innovación (España); Unión Europea. Fondo Europeo de Desarrollo Regional (FEDER/ERDF); University of Alcalá (España)
Numerous studies have focused on the pathophysiological role of amyloid precursor protein (APP) because the proteolytic processing of APP to β-amyloid (Aβ) peptide is a central event in Alzheimer's disease (AD). However, many authors consider that alterations in the physiological functions of APP are likely to play a key role in AD. Previous studies in our laboratory revealed that APP plays an important role in the differentiation of human neural stem cells (hNSCs), favoring glial differentiation (gliogenesis) and preventing their differentiation toward a neuronal phenotype (neurogenesis). In the present study, we have evaluated the effects of APP overexpression in hNSCs at a global gene level by a transcriptomic analysis using the massive RNA sequencing (RNA-seq) technology. Specifically, we have focused on differentially expressed genes that are related to neuronal and glial differentiation processes, as well as on groups of differentially expressed genes associated with different signaling pathways, in order to find a possible interaction between them and APP. Our data indicate a differential expression in genes related to Notch, Wnt, PI3K-AKT, and JAK-STAT signaling, among others. Knowledge of APP biological functions, as well as the possible signaling pathways that could be related to this protein, are essential to advance our understanding of AD.