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Front Immunol ; 13: 773191, 2022.
Article En | MEDLINE | ID: mdl-35371036

Zika virus (ZIKV), despite being discovered six decades earlier, became a major health concern only after an epidemic in French Polynesia and an increase in the number of microcephaly cases in Brazil. Substantial evidence has been found to support the link between ZIKV and neurological complications in infants. The virus targets various cells in the brain, including radial glial cells, neural progenitor cells (NPCs), astrocytes, microglial and glioblastoma stem cells. It affects the brain cells by exploiting different mechanisms, mainly through apoptosis and cell cycle dysregulation. The modulation of host immune response and the inflammatory process has also been demonstrated to play a critical role in ZIKV induced neurological complications. In addition to that, different ZIKV strains have exhibited specific neurotropism and unique molecular mechanisms. This review provides a comprehensive and up-to-date overview of ZIKV-induced neuroimmunopathogenesis by dissecting its main target cells in the brain, and the underlying cellular and molecular mechanisms. We highlighted the roles of the different ZIKV host factors and how they exploit specific host factors through various mechanisms. Overall, it covers key components for understanding the crosstalk between ZIKV and the brain.

Microcephaly , Nervous System Diseases , Neural Stem Cells , Zika Virus Infection , Zika Virus , Brain/pathology , Humans , Microcephaly/pathology , Nervous System Diseases/pathology , Neural Stem Cells/pathology , Zika Virus/physiology
Int J Mol Sci ; 23(4)2022 Feb 09.
Article En | MEDLINE | ID: mdl-35216046

The etiology of juvenile angiofibroma (JA) has been a controversial topic for more than 160 years. Numerous theories have been proposed to explain this rare benign neoplasm arising predominately in adolescent males, focusing mainly on either the vascular or fibrous component. To assess our hypothesis of JA's being a malformation arising from neural crest cells/remnants of the first branchial arch plexus, we performed immunohistochemical analyses of neural crest stem cells (NCSC) and epithelial-mesenchymal transition (EMT) candidates. Immunoexpression of the NCSC marker CD271p75 was observed in all investigated JA's (n = 22), mainly around the pathological vessels. Close to CD271p75-positive cells, high MMP3-staining was also observed. Additionally, from one JA with sufficient material, RT-qPCR identified differences in the expression pattern of PDGFRß, MMP2 and MMP3 in MACS®-separated CD271p75positive vs. CD271p75 negative cell fractions. Our results, together with the consideration of the literature, provide evidence that JA's represent a malformation within the first branchial arch artery/plexus remnants deriving from NCSC. This theory would explain the typical site of tumor origin as well as the characteristic tumor blood supply, whereas the process of EMT provides an explanation for the vascular and fibrous tumor component.

Angiofibroma/pathology , Neural Crest/pathology , Neural Stem Cells/pathology , Adolescent , Adult , Angiofibroma/metabolism , Child , Humans , Male , Middle Aged , Nerve Tissue Proteins/metabolism , Neural Crest/metabolism , Neural Stem Cells/metabolism , Young Adult
PLoS Biol ; 20(1): e3001526, 2022 01.
Article En | MEDLINE | ID: mdl-35085235

The NKCC1 ion transporter contributes to the pathophysiology of common neurological disorders, but its function in microglia, the main inflammatory cells of the brain, has remained unclear to date. Therefore, we generated a novel transgenic mouse line in which microglial NKCC1 was deleted. We show that microglial NKCC1 shapes both baseline and reactive microglia morphology, process recruitment to the site of injury, and adaptation to changes in cellular volume in a cell-autonomous manner via regulating membrane conductance. In addition, microglial NKCC1 deficiency results in NLRP3 inflammasome priming and increased production of interleukin-1ß (IL-1ß), rendering microglia prone to exaggerated inflammatory responses. In line with this, central (intracortical) administration of the NKCC1 blocker, bumetanide, potentiated intracortical lipopolysaccharide (LPS)-induced cytokine levels. In contrast, systemic bumetanide application decreased inflammation in the brain. Microglial NKCC1 KO animals exposed to experimental stroke showed significantly increased brain injury, inflammation, cerebral edema and worse neurological outcome. Thus, NKCC1 emerges as an important player in controlling microglial ion homeostasis and inflammatory responses through which microglia modulate brain injury. The contribution of microglia to central NKCC1 actions is likely to be relevant for common neurological disorders.

Brain Edema/genetics , Brain Injuries/genetics , Microglia/metabolism , Solute Carrier Family 12, Member 2/genetics , Stroke/genetics , Animals , Brain Edema/chemically induced , Brain Edema/metabolism , Brain Edema/pathology , Brain Injuries/chemically induced , Brain Injuries/metabolism , Brain Injuries/pathology , Bumetanide/pharmacology , Embryo, Mammalian , Gene Expression Regulation , Hippocampus/drug effects , Hippocampus/metabolism , Hippocampus/pathology , Inflammasomes/drug effects , Inflammasomes/metabolism , Inflammation , Injections, Intraventricular , Interleukin-1beta/genetics , Interleukin-1beta/metabolism , Lipopolysaccharides/administration & dosage , Male , Mice , Mice, Inbred C57BL , Mice, Knockout , Microglia/drug effects , Microglia/pathology , NLR Family, Pyrin Domain-Containing 3 Protein/genetics , NLR Family, Pyrin Domain-Containing 3 Protein/metabolism , Neural Stem Cells/drug effects , Neural Stem Cells/metabolism , Neural Stem Cells/pathology , Phenotype , Solute Carrier Family 12, Member 2/deficiency , Stroke/chemically induced , Stroke/metabolism , Stroke/pathology
Exp Cell Res ; 412(1): 113007, 2022 03 01.
Article En | MEDLINE | ID: mdl-34990619

Mucopolysaccharidosis type II (MPS II), also known as Hunter syndrome, is a rare, lysosomal disorder caused by mutations in a gene encoding iduronate-2-sulfatase (IDS). IDS deficiency results in an accumulation of glycosaminoglycans (GAGs) and secondary accumulations of other lipids in lysosomes. Symptoms of MPS II include a variety of soft and hard tissue problems, developmental delay, and deterioration of multiple organs. Enzyme replacement therapy is an approved treatment for MPS II, but fails to improve neuronal symptoms. Cell-based neuronal models of MPS II disease are needed for compound screening and drug development for the treatment of the neuronal symptoms in MPS II. In this study, three induced pluripotent stem cell (iPSC) lines were generated from three MPS II patient-derived dermal fibroblast cell lines that were differentiated into neural stem cells and neurons. The disease phenotypes were measured using immunofluorescence staining and Nile red dye staining. In addition, the therapeutic effects of recombinant human IDS enzyme, delta-tocopherol (DT), and hydroxypropyl-beta-cyclodextrin (HPBCD) were determined in the MPS II disease cells. Finally, the neural stem cells from two of the MPS II iPSC lines exhibited typical disease features including a deficiency of IDS activity, abnormal glycosaminoglycan storage, and secondary lipid accumulation. Enzyme replacement therapy partially rescued the disease phenotypes in these cells. DT showed a significant effect in reducing the secondary accumulation of lipids in the MPS II neural stem cells. In contrast, HPBCD displayed limited or no effect in these cells. Our data indicate that these MPS II cells can be used as a cell-based disease model to study disease pathogenesis, evaluate drug efficacy, and screen compounds for drug development.

Induced Pluripotent Stem Cells/drug effects , Induced Pluripotent Stem Cells/metabolism , Mucopolysaccharidosis II/drug therapy , Mucopolysaccharidosis II/metabolism , Neural Stem Cells/drug effects , Neural Stem Cells/metabolism , 2-Hydroxypropyl-beta-cyclodextrin/therapeutic use , Cell Line , Enzyme Replacement Therapy , Glycosaminoglycans/metabolism , Humans , Iduronate Sulfatase/therapeutic use , Induced Pluripotent Stem Cells/pathology , Lipid Metabolism/drug effects , Models, Neurological , Mucopolysaccharidosis II/pathology , Neural Stem Cells/pathology , Phenotype , Recombinant Proteins/therapeutic use , Tocopherols/therapeutic use
Neuron ; 110(1): 12-15, 2022 01 05.
Article En | MEDLINE | ID: mdl-34990576

Dilation of the fluid-filled cerebral ventricles (ventriculomegaly) characterizes hydrocephalus and is frequently seen in autism and schizophrenia. Recent work suggests that the genomic study of congenital hydrocephalus may be unexpectedly fertile ground for revealing insights into neural stem cell regulation, human cerebrocortical development, and pathogenesis of neuropsychiatric disease.

Hydrocephalus , Neural Stem Cells , Cerebral Ventricles , Humans , Hydrocephalus/genetics , Neural Stem Cells/pathology
Cell Tissue Res ; 387(3): 399-414, 2022 Mar.
Article En | MEDLINE | ID: mdl-34820704

Glial scars are a common pathological occurrence in a variety of central nervous system (CNS) diseases and injuries. They are caused after severe damage and consist of reactive glia that form a barrier around the damaged tissue that leads to a non-permissive microenvironment which prevents proper endogenous regeneration. While there are a number of therapies that are able to address some components of disease, there are none that provide regenerative properties. Within the past decade, neural stem cells (NSCs) have been heavily studied due to their potent anti-inflammatory and reparative capabilities in disease and injury. Exogenously applied NSCs have been found to aid in glial scar healing by reducing inflammation and providing cell replacement. However, endogenous NSCs have also been found to contribute to the reactive environment by different means. Further understanding how NSCs can be leveraged to aid in the resolution of the glial scar is imperative in the use of these cells as regenerative therapies. To do so, humanised 3D model systems have been developed to study the development and maintenance of the glial scar. Herein, we explore the current work on endogenous and exogenous NSCs in the glial scar as well as the novel 3D stem cell-based technologies being used to model this pathology in a dish.

Central Nervous System Diseases , Neural Stem Cells , Spinal Cord Injuries , Cicatrix/pathology , Gliosis/pathology , Humans , Neural Stem Cells/pathology , Neuroglia/pathology , Spinal Cord Injuries/therapy
J Stroke Cerebrovasc Dis ; 31(2): 106221, 2022 Feb.
Article En | MEDLINE | ID: mdl-34837757

OBJECTIVE: To assess the potential effect of dl-3-N-butylphthalide (dl-NBP) for the proliferation and differentiation of neural stem cells (NSCs) against hypoxia and the underlying mechanism. MATERIALS AND METHODS: Hippocampal NSCs were obtained from fetal rats. NSCs combined with dl-NBP and single NSCs were cultured. The impact of siRNA-mediated hypoxia-inducible factor-1alpha (HIF-1α) knockdown on NSCs was detected with western blotting (WB) and quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR). Cell-counting kit-8 assay was used for evaluating the viability of NSCs. Levels of HIF-1α protein were measured using WB, and vascular endothelial growth factor (VEGF) expression was quantified using RT-qPCR and enzyme-linked immunosorbent assay. RESULTS: Compared with 7 different concentrations of dl-NBP, 0.25 g/L was determined as the optimal concentration to significantly increase the viability of NSCs (p < 0.001). Dl-NBP can significantly increase the viability of hypoxic NSCs (p < 0.001) and improve the differentiation of hypoxic NSCs into astrocytes (p = 0.001) and oligodendrocytes (p < 0.001). Meanwhile, Dl-NBP can significantly elevate levels of HIF-1α protein (p < 0.001) and VEGF mRNA (p = 0.001) / protein (p < 0.001) in NSCs in the hypoxic environment. However, after transfection with HIF-1α siRNA in NSCs, the viability and differentiation of NSCs was not recovered using dl-NBP under the hypoxic condition, as well as levels of HIF-1α and VEGF. CONCLUSION: Dl-NBP can reverse the weaker proliferation and differentiation power of NSCs in the hypoxic environment. The HIF-1α - VEGF pathway may be implicated in this protective effect of dl-NBP.

Benzofurans , Hypoxia-Inducible Factor 1, alpha Subunit , Hypoxia , Neural Stem Cells , Animals , Benzofurans/pharmacology , Hypoxia/prevention & control , Hypoxia-Inducible Factor 1, alpha Subunit/drug effects , Hypoxia-Inducible Factor 1, alpha Subunit/metabolism , Neural Stem Cells/pathology , Neuroprotective Agents/pharmacology , Rats
Biomaterials ; 280: 121310, 2022 01.
Article En | MEDLINE | ID: mdl-34890972

Radial glia (RG) cells that align in parallel in the embryonic brain are found to be able to guide the directed migration of neurons in response to brain injury. Therefore, biomaterials with aligned architectures are supposed to have positive effects on neural migration and neurogenic differentiation for brain injury repair that are rarely addressed, although they have been widely demonstrated in spinal cord and peripheral nerve system. Here, we present a highly biomimetic scaffold of aligned fibrin hydrogel (AFG) that mimics the oriented structure of RG fibers. Through a combination of histological, behavioral, imaging, and transcriptomic analyses, we demonstrated that transplanting the AFG scaffold into injured cortical brains promotes effective migration, differentiation, and maturation of endogenous neural stem cells, resulting in neurological functional recovery. Therefore, this study will light up a new perspective on applying an aligned scaffold to promote cortical regeneration after injury by inducing endogenous neurogenesis.

Brain Injuries , Neural Stem Cells , Spinal Cord Injuries , Brain/pathology , Brain Injuries/therapy , Cell Differentiation , Humans , Neural Stem Cells/pathology , Neurogenesis , Spinal Cord Injuries/pathology , Spinal Cord Injuries/therapy
Gastroenterology ; 162(1): 179-192.e11, 2022 01.
Article En | MEDLINE | ID: mdl-34425092

BACKGROUND AND AIMS: The enteric nervous system, which regulates many gastrointestinal functions, is derived from neural crest cells (NCCs). Defective NCC migration during embryonic development may lead to enteric neuropathies such as Hirschsprung's disease (hindgut aganglionosis). Sox10 is known to be essential for cell migration but downstream molecular events regulating early NCC migration have not been fully elucidated. This study aimed to determine how Sox10 regulates migration of sacral NCCs toward the hindgut using Dominant megacolon mice, an animal model of Hirschsprung's disease with a Sox10 mutation. METHODS: We used the following: time-lapse live cell imaging to determine the migration defects of mutant sacral NCCs; genome-wide microarrays, site-directed mutagenesis, and whole embryo culture to identify Sox10 targets; and liquid chromatography and tandem mass spectrometry to ascertain downstream effectors of Sox10. RESULTS: Sacral NCCs exhibited retarded migration to the distal hindgut in Sox10-null embryos with simultaneous down-regulated expression of cadherin-19 (Cdh19). Sox10 was found to bind directly to the Cdh19 promoter. Cdh19 knockdown resulted in retarded sacral NCC migration in vitro and ex vivo, whereas re-expression of Cdh19 partially rescued the retarded migration of mutant sacral NCCs in vitro. Cdh19 formed cadherin-catenin complexes, which then bound to filamentous actin of the cytoskeleton during cell migration. CONCLUSIONS: Cdh19 is a direct target of Sox10 during early sacral NCC migration toward the hindgut and forms cadherin-catenin complexes which interact with the cytoskeleton in migrating cells. Elucidation of this novel molecular pathway helps to provide insights into the pathogenesis of enteric nervous system developmental defects.

Cadherins/metabolism , Cell Movement , Enteric Nervous System/metabolism , Hirschsprung Disease/metabolism , Neural Crest/metabolism , Neural Stem Cells/metabolism , Neurogenesis , SOXE Transcription Factors/metabolism , Actin Cytoskeleton/genetics , Actin Cytoskeleton/metabolism , Actin Cytoskeleton/pathology , Animals , Cadherins/genetics , Cells, Cultured , Disease Models, Animal , Embryo Culture Techniques , Enteric Nervous System/abnormalities , Gene Expression Regulation, Developmental , Hirschsprung Disease/genetics , Hirschsprung Disease/pathology , Mice, Inbred C3H , Mice, Inbred C57BL , Mice, Knockout , Neural Crest/abnormalities , Neural Stem Cells/pathology , Protein Binding , SOXE Transcription Factors/genetics , Signal Transduction , Time Factors
Proc Natl Acad Sci U S A ; 119(1)2022 01 04.
Article En | MEDLINE | ID: mdl-34969858

Brain metastases are a leading cause of death in patients with breast cancer. The lack of clinical trials and the presence of the blood-brain barrier limit therapeutic options. Furthermore, overexpression of the human epidermal growth factor receptor 2 (HER2) increases the incidence of breast cancer brain metastases (BCBM). HER2-targeting agents, such as the monoclonal antibodies trastuzumab and pertuzumab, improved outcomes in patients with breast cancer and extracranial metastases. However, continued BCBM progression in breast cancer patients highlighted the need for novel and effective targeted therapies against intracranial metastases. In this study, we engineered the highly migratory and brain tumor tropic human neural stem cells (NSCs) LM008 to continuously secrete high amounts of functional, stable, full-length antibodies against HER2 (anti-HER2Ab) without compromising the stemness of LM008 cells. The secreted anti-HER2Ab impaired tumor cell proliferation in vitro in HER2+ BCBM cells by inhibiting the PI3K-Akt signaling pathway and resulted in a significant benefit when injected in intracranial xenograft models. In addition, dual HER2 blockade using anti-HER2Ab LM008 NSCs and the tyrosine kinase inhibitor tucatinib significantly improved the survival of mice in a clinically relevant model of multiple HER2+ BCBM. These findings provide compelling evidence for the use of HER2Ab-secreting LM008 NSCs in combination with tucatinib as a promising therapeutic regimen for patients with HER2+ BCBM.

Antineoplastic Agents, Immunological/metabolism , Brain Neoplasms , Neoplasms, Experimental , Neural Stem Cells , Oxazoles/pharmacology , Pyridines/pharmacology , Quinazolines/pharmacology , Receptor, ErbB-2 , Animals , Brain Neoplasms/genetics , Brain Neoplasms/metabolism , Brain Neoplasms/pathology , Brain Neoplasms/therapy , Cell Line, Tumor , Humans , Mice , Mice, Nude , Neoplasm Metastasis , Neoplasms, Experimental/genetics , Neoplasms, Experimental/metabolism , Neoplasms, Experimental/pathology , Neoplasms, Experimental/therapy , Neural Stem Cells/metabolism , Neural Stem Cells/pathology , Neural Stem Cells/transplantation , Receptor, ErbB-2/antagonists & inhibitors , Receptor, ErbB-2/metabolism , Xenograft Model Antitumor Assays
Mol Cell ; 82(1): 90-105.e13, 2022 01 06.
Article En | MEDLINE | ID: mdl-34942119

Neurodevelopmental cognitive disorders provide insights into mechanisms of human brain development. Here, we report an intellectual disability syndrome caused by the loss of APC7, a core component of the E3 ubiquitin ligase anaphase promoting complex (APC). In mechanistic studies, we uncover a critical role for APC7 during the recruitment and ubiquitination of APC substrates. In proteomics analyses of the brain from mice harboring the patient-specific APC7 mutation, we identify the chromatin-associated protein Ki-67 as an APC7-dependent substrate of the APC in neurons. Conditional knockout of the APC coactivator protein Cdh1, but not Cdc20, leads to the accumulation of Ki-67 protein in neurons in vivo, suggesting that APC7 is required for the function of Cdh1-APC in the brain. Deregulated neuronal Ki-67 upon APC7 loss localizes predominantly to constitutive heterochromatin. Our findings define an essential function for APC7 and Cdh1-APC in neuronal heterochromatin regulation, with implications for understanding human brain development and disease.

Apc7 Subunit, Anaphase-Promoting Complex-Cyclosome/metabolism , Brain/enzymology , Heterochromatin/metabolism , Intellectual Disability/enzymology , Neural Stem Cells/enzymology , Neurogenesis , Adolescent , Animals , Antigens, CD , Apc7 Subunit, Anaphase-Promoting Complex-Cyclosome/genetics , Behavior, Animal , Brain/growth & development , Cadherins/genetics , Cadherins/metabolism , Cell Line , Child , Child, Preschool , Disease Models, Animal , Female , Heterochromatin/genetics , Humans , Infant , Intellectual Disability/pathology , Intellectual Disability/physiopathology , Intellectual Disability/psychology , Intelligence , Ki-67 Antigen/genetics , Ki-67 Antigen/metabolism , Male , Mice, Inbred C57BL , Mice, Knockout , Mitosis , Mutation , Neural Stem Cells/pathology , Proteolysis , Signal Transduction , Syndrome , Ubiquitination , Young Adult
Comput Math Methods Med ; 2021: 7853335, 2021.
Article En | MEDLINE | ID: mdl-34925543

METHODS: We obtained microarray data (GSE116726, GSE67566) from Gene Expression Omnibus database, and differential expression level of ncRNA in nucleus pulposus (NP) tissues of IDD patients was analyzed. The potential circRNA-miRNA-mRNA regulatory network was analyzed by starBase. The effect of the interaction between hsa_circ_0001658, hsa-miR-181c-5p, and FAS on the proliferation and apoptosis of human neural progenitor cells (hNPCs) was studied. RESULTS: hsa_circ_0001658 was significantly upregulated (logFC > 2.0 and adj.P.Val < 0.01) in the NP tissues of IDD patients, and hsa-miR-181c-5p expression was downregulated (logFC < -2.0 and adj.P.Val < 0.01). Silencing of hsa-miR-181c-5p or overexpression of hsa_circ_0001658 inhibited the proliferation of hNPCs and promoted their apoptosis. hsa_circ_0001658 acted as a sponge of hsa-miR-181c-5p. hsa-miR-181c-5p downregulated the expression of Fas cell surface death receptor (FAS), promoted the proliferation, and inhibited the apoptosis of hNPCs. hsa_circ_0001658 functioned in hNPCs through targeting hsa-miR-181c-5p/FAS. CONCLUSION: Circular RNA hsa_circ_0001658 inhibits IDD development by regulating hsa-miR-181c-5p/FAS. It is expected to be a potential target for the therapy of IDD.

Intervertebral Disc Degeneration/genetics , MicroRNAs/genetics , RNA, Circular/genetics , fas Receptor/genetics , Apoptosis/genetics , Cell Proliferation/genetics , Cells, Cultured , Computational Biology , Databases, Genetic/statistics & numerical data , Gene Expression , Gene Regulatory Networks , Gene Silencing , Humans , Intervertebral Disc Degeneration/pathology , Intervertebral Disc Degeneration/prevention & control , MicroRNAs/metabolism , Neural Stem Cells/metabolism , Neural Stem Cells/pathology , Nucleus Pulposus/metabolism , Nucleus Pulposus/pathology , RNA, Circular/metabolism , RNA, Messenger/genetics , RNA, Messenger/metabolism
Genes (Basel) ; 12(12)2021 11 27.
Article En | MEDLINE | ID: mdl-34946850

Autism spectrum disorder (ASD) is a neurodevelopmental disorder that impedes patients' cognition, social, speech and communication skills. ASD is highly heterogeneous with a variety of etiologies and clinical manifestations. The prevalence rate of ASD increased steadily in recent years. Presently, molecular mechanisms underlying ASD occurrence and development remain to be elucidated. Here, we integrated multi-layer genomics data to investigate the transcriptome and pathway dysregulations in ASD development. The RNA sequencing (RNA-seq) expression profiles of induced pluripotent stem cells (iPSCs), neural progenitor cells (NPCs) and neuron cells from ASD and normal samples were compared in our study. We found that substantially more genes were differentially expressed in the NPCs than the iPSCs. Consistently, gene set variation analysis revealed that the activity of the known ASD pathways in NPCs and neural cells were significantly different from the iPSCs, suggesting that ASD occurred at the early stage of neural system development. We further constructed comprehensive brain- and neural-specific regulatory networks by incorporating transcription factor (TF) and gene interactions with long 5 non-coding RNA(lncRNA) and protein interactions. We then overlaid the transcriptomes of different cell types on the regulatory networks to infer the regulatory cascades. The variations of the regulatory cascades between ASD and normal samples uncovered a set of novel disease-associated genes and gene interactions, particularly highlighting the functional roles of ELF3 and the interaction between STAT1 and lncRNA ELF3-AS 1 in the disease development. These new findings extend our understanding of ASD and offer putative new therapeutic targets for further studies.

Autism Spectrum Disorder/genetics , Gene Regulatory Networks/genetics , Neurons/pathology , Autism Spectrum Disorder/pathology , Gene Expression Profiling/methods , Gene Expression Regulation/genetics , Humans , Induced Pluripotent Stem Cells/pathology , Neural Stem Cells/pathology , Organogenesis/genetics , Sequence Analysis, RNA/methods , Transcription Factors/genetics , Transcriptome/genetics
Cells ; 10(12)2021 12 09.
Article En | MEDLINE | ID: mdl-34943986

Parkinson's Disease (PD) is a widespread severe neurodegenerative disease that is characterized by pronounced deficiency of the dopaminergic system and disruption of the function of other neuromodulator systems. Although heritable genetic factors contribute significantly to PD pathogenesis, only a small percentage of sporadic cases of PD can be explained using known genetic risk factors. Due to that, it could be inferred that changes in gene expression could be important for explaining a significant percentage of PD cases. One of the ways to investigate such changes, while minimizing the effect of genetic factors on experiment, are the study of PD discordant monozygotic twins. In the course of the analysis of transcriptome data obtained from IPSC and NPCs, 20 and 1906 differentially expressed genes were identified respectively. We have observed an overexpression of TNF in NPC cultures, derived from twin with PD. Through investigation of gene interactions and gene involvement in biological processes, we have arrived to a hypothesis that TNF could play a crucial role in PD-related changes occurring in NPC derived from twins with PD, and identified INHBA, WNT7A and DKK1 as possible downstream effectors of TNF.

Induced Pluripotent Stem Cells/metabolism , Neurodegenerative Diseases/genetics , Parkinson Disease/genetics , Transcriptome/genetics , Aged , Cell Differentiation , Dopamine/genetics , Female , Gene Expression Profiling , Humans , Induced Pluripotent Stem Cells/pathology , Neural Stem Cells/metabolism , Neural Stem Cells/pathology , Neurodegenerative Diseases/pathology , Neurons/metabolism , Parkinson Disease/pathology , Twins, Monozygotic/genetics
Sci Rep ; 11(1): 22891, 2021 11 24.
Article En | MEDLINE | ID: mdl-34819604

The balances between NSCs growth and differentiation, and between glial and neuronal differentiation play a key role in brain regeneration after any pathological conditions. It is well known that the nervous tissue shows a poor recovery after injury due to the factors present in the wounded microenvironment, particularly inflammatory factors, that prevent neuronal differentiation. Thus, it is essential to generate a favourable condition for NSCs and conduct them to differentiate towards functional neurons. Here, we show that neuroinflammation has no effect on NSCs proliferation but induces an aberrant neuronal differentiation that gives rise to dystrophic, non-functional neurons. This is perhaps the initial step of brain failure associated to many neurological disorders. Interestingly, we demonstrate that phosphatidylcholine (PtdCho)-enriched media enhances neuronal differentiation even under inflammatory stress by modifying the commitment of post-mitotic cells. The pro-neurogenic effect of PtdCho increases the population of healthy normal neurons. In addition, we provide evidences that this phospholipid ameliorates the damage of neurons and, in consequence, modulates neuronal plasticity. These results contribute to our understanding of NSCs behaviour under inflammatory conditions, opening up new venues to improve neurogenic capacity in the brain.

Cell Plasticity/drug effects , Neural Stem Cells/drug effects , Neurogenesis/drug effects , Phosphatidylcholines/pharmacology , Synapses/drug effects , Animals , Cell Proliferation/drug effects , Inflammation Mediators/metabolism , Macrophages/metabolism , Mice , Mice, Inbred C57BL , Neural Stem Cells/metabolism , Neural Stem Cells/pathology , /pathology , Phenotype , RAW 264.7 Cells , Synapses/metabolism , Synapses/pathology
Sci Rep ; 11(1): 22904, 2021 11 25.
Article En | MEDLINE | ID: mdl-34824314

In Alzheimer´s disease (AD) there is a reduction in hippocampal neurogenesis that has been associated to cognitive deficits. Previously we showed that Andrographolide (ANDRO), the main bioactive component of Andrographis paniculate, induces proliferation in the hippocampus of the APPswe/PSEN1ΔE9 (APP/PS1) mouse model of AD as assessed by staining with the mitotic marker Ki67. Here, we further characterized the effect of ANDRO on hippocampal neurogenesis in APP/PS1 mice and evaluated the contribution of this process to the cognitive effect of ANDRO. Treatment of 8-month-old APP/PS1 mice with ANDRO for 4 weeks increased proliferation in the dentate gyrus as evaluated by BrdU incorporation. Although ANDRO had no effect on neuronal differentiation of newborn cells, it strongly increased neural progenitors, neuroblasts and newborn immature neurons, cell populations that were decreased in APP/PS1 mice compared to age-matched wild-type mice. ANDRO had no effect on migration or in total dendritic length, arborization and orientation of immature neurons, suggesting no effects on early morphological development of newborn neurons. Finally, ANDRO treatment improved the performance of APP/PS1 mice in the object location memory task. This effect was not completely prevented by co-treatment with the anti-mitotic drug TMZ, suggesting that other effects of ANDRO in addition to the increase in neurogenesis might underlie the observed cognitive improvement. Altogether, our data indicate that in APP/PS1 mice ANDRO stimulates neurogenesis in the hippocampus by inducing proliferation of neural precursor cells and improves spatial memory performance.

Alzheimer Disease/drug therapy , Behavior, Animal/drug effects , Cell Proliferation/drug effects , Dentate Gyrus/drug effects , Diterpenes/pharmacology , Neural Stem Cells/drug effects , Neurogenesis/drug effects , Neurons/drug effects , Nootropic Agents/pharmacology , Spatial Memory/drug effects , Alzheimer Disease/genetics , Alzheimer Disease/pathology , Alzheimer Disease/psychology , Amyloid beta-Protein Precursor/genetics , Animals , Dentate Gyrus/pathology , Disease Models, Animal , Female , Genetic Predisposition to Disease , Mice, Transgenic , Neural Stem Cells/pathology , Neurons/pathology , Presenilin-1/genetics
Cell Rep ; 37(5): 109912, 2021 11 02.
Article En | MEDLINE | ID: mdl-34731622

Fetal growth restriction (FGR) increases the risk for impaired cognitive function later in life. However, the precise mechanisms remain elusive. Using dexamethasone-induced FGR and protein restriction-influenced FGR mouse models, we observe learning and memory deficits in adult FGR offspring. FGR induces decreased hippocampal neurogenesis from the early post-natal period to adulthood by reducing the proliferation of neural stem cells (NSCs). We further find a persistent decrease of Tet1 expression in hippocampal NSCs of FGR mice. Mechanistically, Tet1 downregulation results in hypermethylation of the Dll3 and Notch1 promoters and inhibition of Notch signaling, leading to reduced NSC proliferation. Overexpression of Tet1 activates Notch signaling, offsets the decline in neurogenesis, and enhances learning and memory abilities in FGR offspring. Our data indicate that a long-term decrease in Tet1/Notch signaling in hippocampal NSCs contributes to impaired neurogenesis following FGR and could serve as potential targets for the intervention of FGR-related cognitive disorders.

Behavior, Animal , Cognition , DNA-Binding Proteins/metabolism , Fetal Growth Retardation/metabolism , Hippocampus/metabolism , Neural Stem Cells/metabolism , Neurogenesis , Proto-Oncogene Proteins/metabolism , Animals , Cell Proliferation , Cells, Cultured , DNA Methylation , DNA-Binding Proteins/genetics , Disease Models, Animal , Epigenesis, Genetic , Female , Fetal Growth Retardation/physiopathology , Fetal Growth Retardation/psychology , Hippocampus/physiopathology , Intracellular Signaling Peptides and Proteins/genetics , Intracellular Signaling Peptides and Proteins/metabolism , Male , Membrane Proteins/genetics , Membrane Proteins/metabolism , Memory , Mice, Inbred C57BL , Neural Stem Cells/pathology , Pregnancy , Prenatal Exposure Delayed Effects , Proto-Oncogene Proteins/genetics , Receptor, Notch1/genetics , Receptor, Notch1/metabolism , Signal Transduction
Toxicol Appl Pharmacol ; 433: 115792, 2021 12 15.
Article En | MEDLINE | ID: mdl-34742744

Concurrent with the '3R' principle, the embryonic stem cell test (EST) using mouse embryonic stem cells, developed in 2000, remains the solely accepted in vitro method for embryotoxicity testing. However, the scope and implementation of EST for embryotoxicity screening, compliant with regulatory requirements, are limited. This is due to its technical complexity, long testing period, labor-intensive methodology, and limited endpoint data, leading to misclassification of embryotoxic potential. In this study, we used human induced pluripotent stem cell (hiPSC)-derived embryoid bodies (EB) as an in vitro model to investigate the embryotoxic effects of a carefully selected set of pharmacological compounds. Morphology, viability, and differentiation potential were investigated after exposing EBs to folic acid, all-trans-retinoic acid, dexamethasone, and valproic acid for 15 days. The results showed that the compounds differentially repressed cell growth, compromised morphology, and triggered apoptosis in the EBs. Further, transcriptomics was employed to compare subtle temporal changes between treated and untreated cultures. Gene ontology and pathway analysis revealed that dysregulation of a large number of genes strongly correlated with impaired neuroectoderm and cardiac mesoderm formation. This aberrant gene expression pattern was associated with several disorders of the brain like mental retardation, multiple sclerosis, stroke and of the heart like dilated cardiomyopathy, ventricular tachycardia, and ventricular arrhythmia. Lastly, these in vitro findings were validated using in ovo chick embryo model. Taken together, pharmacological compound or drug-induced defective EB development from hiPSCs could potentially be used as a suitable in vitro platform for embryotoxicity screening.

Cell Differentiation/drug effects , Embryoid Bodies/drug effects , Gene Expression Profiling , Induced Pluripotent Stem Cells/drug effects , Myocytes, Cardiac/drug effects , Neural Stem Cells/drug effects , Teratogens/toxicity , Toxicity Tests , Transcriptome/drug effects , Animals , Apoptosis/drug effects , Cell Line , Cell Lineage , Chick Embryo , Dexamethasone/toxicity , Dose-Response Relationship, Drug , Embryoid Bodies/metabolism , Embryoid Bodies/pathology , Gene Expression Regulation, Developmental/drug effects , Humans , Induced Pluripotent Stem Cells/metabolism , Induced Pluripotent Stem Cells/pathology , Inhibitory Concentration 50 , Myocytes, Cardiac/metabolism , Myocytes, Cardiac/pathology , Neural Stem Cells/metabolism , Neural Stem Cells/pathology , Neurogenesis/drug effects , Risk Assessment , Tretinoin/toxicity , Valproic Acid/toxicity
Cells ; 10(11)2021 11 22.
Article En | MEDLINE | ID: mdl-34831487

Diabetic retinopathy is a frequent complication of longstanding diabetes, which comprises a complex interplay of microvascular abnormalities and neurodegeneration. Zebrafish harboring a homozygous mutation in the pancreatic transcription factor pdx1 display a diabetic phenotype with survival into adulthood, and are therefore uniquely suitable among zebrafish models for studying pathologies associated with persistent diabetic conditions. We have previously shown that, starting at three months of age, pdx1 mutants exhibit not only vascular but also neuro-retinal pathologies manifesting as photoreceptor dysfunction and loss, similar to human diabetic retinopathy. Here, we further characterize injury and regenerative responses and examine the effects on progenitor cell populations. Consistent with a negative impact of hyperglycemia on neurogenesis, stem cells of the ciliary marginal zone show an exacerbation of aging-related proliferative decline. In contrast to the robust Müller glial cell proliferation seen following acute retinal injury, the pdx1 mutant shows replenishment of both rod and cone photoreceptors from slow-cycling, neurod-expressing progenitors which first accumulate in the inner nuclear layer. Overall, we demonstrate a diabetic retinopathy model which shows pathological features of the human disease evolving alongside an ongoing restorative process that replaces lost photoreceptors, at the same time suggesting an unappreciated phenotypic continuum between multipotent and photoreceptor-committed progenitors.

Hyperglycemia/pathology , Neural Stem Cells/pathology , Retina/pathology , Aging/pathology , Animals , Basic Helix-Loop-Helix Transcription Factors/metabolism , Cell Death , Cell Proliferation , Chronic Disease , Ependymoglial Cells/pathology , Green Fluorescent Proteins/metabolism , Homeodomain Proteins/genetics , Models, Biological , Mutation/genetics , Nerve Tissue Proteins/metabolism , PAX6 Transcription Factor/metabolism , Photoreceptor Cells/metabolism , Photoreceptor Cells/pathology , Receptors, Notch/metabolism , Retina/immunology , Signal Transduction , Trans-Activators/genetics , Zebrafish
Int J Mol Sci ; 22(21)2021 Oct 30.
Article En | MEDLINE | ID: mdl-34769232

Changes in adult hippocampal cell proliferation and genesis have been largely implicated in depression and antidepressant action, though surprisingly, the underlying cell cycle mechanisms are largely undisclosed. Using both an in vivo unpredictable chronic mild stress (uCMS) rat model of depression and in vitro rat hippocampal-derived neurosphere culture approaches, we aimed to unravel the cell cycle mechanisms regulating hippocampal cell proliferation and genesis in depression and after antidepressant treatment. We show that the hippocampal dentate gyrus (hDG) of uCMS animals have less proliferating cells and a decreased proportion of cells in the G2/M phase, suggesting a G1 phase arrest; this is accompanied by decreased levels of cyclin D1, E, and A expression. Chronic fluoxetine treatment reversed the G1 phase arrest and promoted an up-regulation of cyclin E. In vitro, dexamethasone (DEX) decreased cell proliferation, whereas the administration of serotonin (5-HT) reversed it. DEX also induced a G1-phase arrest and decreased cyclin D1 and D2 expression levels while increasing p27. Additionally, 5-HT treatment could partly reverse the G1-phase arrest and restored cyclin D1 expression. We suggest that the anti-proliferative actions of chronic stress in the hDG result from a glucocorticoid-mediated G1-phase arrest in the progenitor cells that is partly mediated by decreased cyclin D1 expression which may be overcome by antidepressant treatment.

Cyclins/metabolism , Depression , Fluoxetine/pharmacology , Hippocampus/metabolism , Neural Stem Cells/metabolism , Animals , Depression/drug therapy , Depression/metabolism , Depression/pathology , Dexamethasone/pharmacology , Disease Models, Animal , G1 Phase Cell Cycle Checkpoints/drug effects , Hippocampus/pathology , Male , Neural Stem Cells/pathology , Rats , Serotonin/pharmacology