Articles de revues : « IPSC-Derived neural models »

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Amalakanti, *Sridhar, Vijaya Chandra Reddy Avula et Sachin Singh. « SYSTEMATIC REVIEW OF INDUCED PLURIPOTENT STEM CELL THERAPY IN TRAUMATIC BRAIN INJURY ». International Journal of Neuropsychopharmacology 28, Supplement_1 (février 2025) : i364—i365. https://doi.org/10.1093/ijnp/pyae059.649.

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Abstract Background Traumatic brain injury (TBI) is a major global health problem with limited treatment options. Induced pluripotent stem cells (iPSCs) have emerged as a promising therapy for neural regeneration and repair after TBI. Aims & Objectives This systematic review aimed to evaluate preclinical studies on the efficacy of iPSC- based therapies for functional recovery after TBI. Method From, PubMed, Ovid Medline, Cochrane Library, SCOPUS, and Web of Science 102 studies were found for studies on iPSCs in TBI animal models. Included studies (n=9) used neural stem cells derived from iPSCs transplanted into rodent models of TBI. Outcome measures were brain injury volume, functional recovery, and tissue repair biomarkers. Study quality was assessed using a 10-point scale. Results The majority of studies showed significant improvement in motor function, cognition, and social behavior with iPSC therapy. Transplanted iPSC-neural stem cells migrated to injury sites, differentiated into neurons and glia, reduced lesion size, and increased neural repair markers. Higher quality scores were noted for studies reporting randomization, blinded assessment, and temperature control. Discussion & Conclusions Preclinical evidence demonstrates the potential of iPSC-derived neural stem cell transplantation to improve functional outcomes after TBI through neuroregenerative effects. Further research is warranted to evaluate safety, optimize protocols, and translate findings to clinical trials for TBI patients. iPSC-based therapies may offer new hope for recovery after this devastating injury.

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Yang, Guang, Hyenjong Hong, April Torres, Kristen Malloy, Gourav Choudhury, Jeffrey Kim et Marcel Daadi. « Standards for Deriving Nonhuman Primate-Induced Pluripotent Stem Cells, Neural Stem Cells and Dopaminergic Lineage ». International Journal of Molecular Sciences 19, n^o 9 (17 septembre 2018) : 2788. http://dx.doi.org/10.3390/ijms19092788.

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Humans and nonhuman primates (NHP) are similar in behavior and in physiology, specifically the structure, function, and complexity of the immune system. Thus, NHP models are desirable for pathophysiology and pharmacology/toxicology studies. Furthermore, NHP-derived induced pluripotent stem cells (iPSCs) may enable transformative developmental, translational, or evolutionary studies in a field of inquiry currently hampered by the limited availability of research specimens. NHP-iPSCs may address specific questions that can be studied back and forth between in vitro cellular assays and in vivo experimentations, an investigational process that in most cases cannot be performed on humans because of safety and ethical issues. The use of NHP model systems and cell specific in vitro models is evolving with iPSC-based three-dimensional (3D) cell culture systems and organoids, which may offer reliable in vitro models and reduce the number of animals used in experimental research. IPSCs have the potential to give rise to defined cell types of any organ of the body. However, standards for deriving defined and validated NHP iPSCs are missing. Standards for deriving high-quality iPSC cell lines promote rigorous and replicable scientific research and likewise, validated cell lines reduce variability and discrepancies in results between laboratories. We have derived and validated NHP iPSC lines by confirming their pluripotency and propensity to differentiate into all three germ layers (ectoderm, mesoderm, and endoderm) according to standards and measurable limits for a set of marker genes. The iPSC lines were characterized for their potential to generate neural stem cells and to differentiate into dopaminergic neurons. These iPSC lines are available to the scientific community. NHP-iPSCs fulfill a unique niche in comparative genomics to understand gene regulatory principles underlying emergence of human traits, in infectious disease pathogenesis, in vaccine development, and in immunological barriers in regenerative medicine.

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Supakul, Sopak, Chisato Oyama, Yuki Hatakeyama, Sumihiro Maeda et Hideyuki Okano. « Estradiol enhanced neuronal plasticity and ameliorated astrogliosis in human iPSC-derived neural models ». Regenerative Therapy 25 (mars 2024) : 250–63. http://dx.doi.org/10.1016/j.reth.2023.12.018.

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Liu, Sijun, Yuying Zhao, Xiaoying Su, Chengcheng Zhou, Peifen Yang, Qiusan Lin, Shijun Li et al. « Reconstruction of Alzheimer’s Disease Cell Model In Vitro via Extracted Peripheral Blood Molecular Cells from a Sporadic Patient ». Stem Cells International 2020 (18 décembre 2020) : 1–10. http://dx.doi.org/10.1155/2020/8897494.

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The establishment of human-induced pluripotent stem cell (iPSC) models from sporadic Alzheimer’s disease (sAD) patients is necessary and could potentially benefit research into disease etiology and therapeutic strategies. However, the development of sAD iPSC models is still limited due to the multifactorial nature of the disease. Here, we extracted peripheral blood mononuclear cells (PBMCs) from a patient with sAD and induced them into iPSC by introducing the Sendai virus expressing Oct3/4, Sox2, c-Myc, and Klf4, which were subsequently induced into neural cells to build the cell model of AD. Using alkaline phosphatase staining, immunofluorescence staining, karyotype analysis, reverse transcription-polymerase chain reaction (RT-PCR), and teratoma formation in vitro, we demonstrated that the iPSC derived from PMBCs (PBMC-iPSC) had a normal karyotype and potential to differentiate into three embryonic layers. Immunofluorescence staining and quantitative real-time polymerase chain reaction (qPCR) suggested that PBMC-iPSCs were successfully differentiated into neural cells. Detection of beta-amyloid protein oligomer (AβO), beta-amyloid protein 1-40 (Aβ 1-40), and beta-amyloid protein 1-42 (Aβ 1-42) indicated that the AD cell model was satisfactorily constructed in vitro. In conclusion, this study has successfully generated an AD cell model with pathological features of beta-amyloid peptide deposition using PBMC from a patient with sAD.

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Barak, Martin, Veronika Fedorova, Veronika Pospisilova, Jan Raska, Simona Vochyanova, Jiri Sedmik, Hana Hribkova, Hana Klimova, Tereza Vanova et Dasa Bohaciakova. « Human iPSC-Derived Neural Models for Studying Alzheimer’s Disease : from Neural Stem Cells to Cerebral Organoids ». Stem Cell Reviews and Reports 18, n^o 2 (février 2022) : 792–820. http://dx.doi.org/10.1007/s12015-021-10254-3.

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AbstractDuring the past two decades, induced pluripotent stem cells (iPSCs) have been widely used to study mechanisms of human neural development, disease modeling, and drug discovery in vitro. Especially in the field of Alzheimer’s disease (AD), where this treatment is lacking, tremendous effort has been put into the investigation of molecular mechanisms behind this disease using induced pluripotent stem cell-based models. Numerous of these studies have found either novel regulatory mechanisms that could be exploited to develop relevant drugs for AD treatment or have already tested small molecules on in vitro cultures, directly demonstrating their effect on amelioration of AD-associated pathology. This review thus summarizes currently used differentiation strategies of induced pluripotent stem cells towards neuronal and glial cell types and cerebral organoids and their utilization in modeling AD and potential drug discovery. Graphical abstract

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Costamagna, Gianluca, Giacomo Pietro Comi et Stefania Corti. « Advancing Drug Discovery for Neurological Disorders Using iPSC-Derived Neural Organoids ». International Journal of Molecular Sciences 22, n^o 5 (6 mars 2021) : 2659. http://dx.doi.org/10.3390/ijms22052659.

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In the last decade, different research groups in the academic setting have developed induced pluripotent stem cell-based protocols to generate three-dimensional, multicellular, neural organoids. Their use to model brain biology, early neural development, and human diseases has provided new insights into the pathophysiology of neuropsychiatric and neurological disorders, including microcephaly, autism, Parkinson’s disease, and Alzheimer’s disease. However, the adoption of organoid technology for large-scale drug screening in the industry has been hampered by challenges with reproducibility, scalability, and translatability to human disease. Potential technical solutions to expand their use in drug discovery pipelines include Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) to create isogenic models, single-cell RNA sequencing to characterize the model at a cellular level, and machine learning to analyze complex data sets. In addition, high-content imaging, automated liquid handling, and standardized assays represent other valuable tools toward this goal. Though several open issues still hamper the full implementation of the organoid technology outside academia, rapid progress in this field will help to prompt its translation toward large-scale drug screening for neurological disorders.

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Hunt, Jack F. V., Meng Li, Ryan Risgaard, Gene E. Ananiev, Scott Wildman, Fan Zhang, Tim S. Bugni, Xinyu Zhao et Anita Bhattacharyya. « High Throughput Small Molecule Screen for Reactivation of FMR1 in Fragile X Syndrome Human Neural Cells ». Cells 11, n^o 1 (27 décembre 2021) : 69. http://dx.doi.org/10.3390/cells11010069.

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Fragile X syndrome (FXS) is the most common inherited cause of autism and intellectual disability. The majority of FXS cases are caused by transcriptional repression of the FMR1 gene due to epigenetic changes that are not recapitulated in current animal disease models. FXS patient induced pluripotent stem cell (iPSC)-derived gene edited reporter cell lines enable novel strategies to discover reactivators of FMR1 expression in human cells on a much larger scale than previously possible. Here, we describe the workflow using FXS iPSC-derived neural cell lines to conduct a massive, unbiased screen for small molecule activators of the FMR1 gene. The proof-of-principle methodology demonstrates the utility of human stem-cell-based methodology for the untargeted discovery of reactivators of the human FMR1 gene that can be applied to other diseases.

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Csöbönyeiová, Mária, Štefan Polák et L’uboš Danišovič. « Toxicity testing and drug screening using iPSC-derived hepatocytes, cardiomyocytes, and neural cells ». Canadian Journal of Physiology and Pharmacology 94, n^o 7 (juillet 2016) : 687–94. http://dx.doi.org/10.1139/cjpp-2015-0459.

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Unexpected toxicity in areas such as cardiotoxicity, hepatotoxicity, and neurotoxicity is a serious complication of clinical therapy and one of the key causes for failure of promising drug candidates in development. Animal studies have been widely used for toxicology research to provide preclinical security evaluation of various therapeutic agents under development. Species differences in drug penetration of the blood–brain barrier, drug metabolism, and related toxicity contribute to failure of drug trials from animal models to human. The existing system for drug discovery has relied on immortalized cell lines, animal models of human disease, and clinical trials in humans. Moreover, drug candidates that are passed as being safe in the preclinical stage often show toxic effects during the clinical stage. Only around 16% drugs are approved for human use. Research on induced pluripotent stem cells (iPSCs) promises to enhance drug discovery and development by providing simple, reproducible, and economically effective tools for drug toxicity screening under development and, on the other hand, for studying the disease mechanism and pathways. In this review, we provide an overview of basic information about iPSCs, and discuss efforts aimed at the use of iPSC-derived hepatocytes, cardiomyocytes, and neural cells in drug discovery and toxicity testing.

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Fernández-Santiago, Rubén, et Mario Ezquerra. « Epigenetic Research of Neurodegenerative Disorders Using Patient iPSC-Based Models ». Stem Cells International 2016 (2016) : 1–16. http://dx.doi.org/10.1155/2016/9464591.

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Epigenetic mechanisms play a role in human disease but their involvement in pathologies from the central nervous system has been hampered by the complexity of the brain together with its unique cellular architecture and diversity. Until recently, disease targeted neural types were only available as postmortem materials after many years of disease evolution. Current in vitro systems of induced pluripotent stem cells (iPSCs) generated by cell reprogramming of somatic cells from patients have provided valuable disease models recapitulating key pathological molecular events. Yet whether cell reprogramming on itself implies a truly epigenetic reprogramming, the epigenetic mechanisms governing this process are only partially understood. Moreover, elucidating epigenetic regulation using patient-specific iPSC-derived neural models is expected to have a great impact to unravel the pathophysiology of neurodegenerative diseases and to hopefully expand future therapeutic possibilities. Here we will critically review current knowledge of epigenetic involvement in neurodegenerative disorders focusing on the potential of iPSCs as a promising tool for epigenetic research of these diseases.

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Tamura, Ryota, Masahiro Yo, Hiroyuki Miyoshi, Oltea Sampetrean, Hideyuki Saya, Hideyuki Okano et Masahiro Toda. « ET-1 STEM CELL-BASED GENE THERAPY FOR MALIGNANT GLIOMA USING GENOME-EDITED HUMAN INDUCED PLURIPOTENT STEM CELLS ». Neuro-Oncology Advances 4, Supplement_3 (1 décembre 2022) : iii4—iii5. http://dx.doi.org/10.1093/noajnl/vdac167.015.

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Abstract Glioblastoma is the most aggressive primary brain tumor, and is characterized by diffuse infiltration into the normal brain parenchyma. New therapeutic approaches targeting invasive biological behaviour are warranted. In the present study, we show that neural stem cells (NSCs) derived from CRISRP/Cas9-edited induced pluripotent stem cells (iPSCs) have high tumor-trophic migratory capacity and stable constitutive therapeutic transgene expression, which leads to strong anti-tumor effects against glioma stem cell (GSC) models. The present study provides answers to some important research questions associated with stem cell-based gene therapy. First, the tumor-trophic migratory capacities of human iPSC-derived NSCs (iPSC-NSCs), fetal NSCs, and mesenchymal stem cells (MSCs) were quantitatively evaluated by spatiotemporal methodologies. We demonstrated that iPSC-NSCs have a higher tumor-trophic migratory capacity than MSCs in the brain. Self-repulsive action and pathotropism were important for the migration of iPSC-NSCs: ephrin ligand/receptor mediated repulsion of iPSC-NSCs and CXCL12-CXCR4 interactions between GSCs and iPSC-NSCs. Second, a prodrug converting enzyme fusion gene was selected as a therapeutic gene in human iPSCs. In general, stable constitutive transgene expression by viral vectors was difficult in human iPSCs. Furthermore, viral vectors integrate randomly into the host genome, which raises concerns about transgene silencing, insertional mutagenesis, and oncogene activation. In the present study, several common insertion sites including GAPDH, ACTB, and AAVS1, were compared. The most appropriate gene locus that achieved stable constitutive transgene expression was determined via CRISPR/Cas9-mediated genome editing. Third, we revealed the novel mechanism of action using iPSC-NSCs expressing CD-UPRT, in which ferroptosis was associated with enhanced anti-tumor immune responses. We demonstrated that the established iPSC-NSCs had strong therapeutic efficacy in GSC animal models. Finally, predictive biomarkers for the efficacy of the present treatment strategy were established. We will conduct a clinical trial of this treatment strategy. This research concept can disseminate biological, medical and engineering advances.

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Tamura, Ryota, Masahiro Yo, Ryotaro Imai, Hideyuki Okano et Masahiro Toda. « 10000-SPE-1 STEM CELL-BASED GENE THERAPY FOR MALIGNANT GLIOMA USING GENOME-EDITED HUMAN INDUCED PLURIPOTENT STEM CELLS ». Neuro-Oncology Advances 5, Supplement_5 (1 décembre 2023) : v1. http://dx.doi.org/10.1093/noajnl/vdad141.002.

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Abstract Glioblastoma is the most aggressive primary brain tumor, and is characterized by diffuse infiltration into the normal brain parenchyma. New therapeutic approaches targeting invasive biological behaviour are warranted. In the present study, we show that neural stem cells (NSCs) derived from CRISRP/Cas9-edited induced pluripotent stem cells (iPSCs) have high tumor-trophic migratory capacity and stable constitutive therapeutic transgene expression, which leads to strong anti-tumor effects against glioma stem cell (GSC) models. The present study provides answers to some important research questions associated with stem cell-based gene therapy. First, the tumor-trophic migratory capacities of human iPSC-derived NSCs (iPSC-NSCs), fetal NSCs, and mesenchymal stem cells (MSCs) were quantitatively evaluated by spatiotemporal methodologies. We demonstrated that iPSC-NSCs have a higher tumor-trophic migratory capacity than MSCs in the brain. Self-repulsive action and pathotropism were important for the migration of iPSC-NSCs: ephrin ligand/receptor mediated repulsion of iPSC-NSCs and CXCL12-CXCR4 interactions between GSCs and iPSC-NSCs. Second, a prodrug converting enzyme fusion gene was selected as a therapeutic gene in human iPSCs. In general, stable constitutive transgene expression by viral vectors was difficult in human iPSCs. Furthermore, viral vectors integrate randomly into the host genome, which raises concerns about transgene silencing, insertional mutagenesis, and oncogene activation. In the present study, several common insertion sites including GAPDH, ACTB, and AAVS1, were compared. The most appropriate gene locus that achieved stable constitutive transgene expression was determined via CRISPR/Cas9-mediated genome editing. Third, we revealed the novel mechanism of action using iPSC-NSCs expressing CD-UPRT, in which ferroptosis was associated with enhanced anti-tumor immune responses. We demonstrated that the established iPSC-NSCs had strong therapeutic efficacy in GSC animal models. Finally, predictive biomarkers for the efficacy of the present treatment strategy were established. We will conduct a clinical trial of this treatment strategy. This research concept can disseminate biological, medical and engineering advances.

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Kiaee, Kiavash, Yasamin A. Jodat, Nicole J. Bassous, Navneet Matharu et Su Ryon Shin. « Transcriptomic Mapping of Neural Diversity, Differentiation and Functional Trajectory in iPSC-Derived 3D Brain Organoid Models ». Cells 10, n^o 12 (5 décembre 2021) : 3422. http://dx.doi.org/10.3390/cells10123422.

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Experimental models of the central nervous system (CNS) are imperative for developmental and pathophysiological studies of neurological diseases. Among these models, three-dimensional (3D) induced pluripotent stem cell (iPSC)-derived brain organoid models have been successful in mitigating some of the drawbacks of 2D models; however, they are plagued by high organoid-to-organoid variability, making it difficult to compare specific gene regulatory pathways across 3D organoids with those of the native brain. Single-cell RNA sequencing (scRNA-seq) transcriptome datasets have recently emerged as powerful tools to perform integrative analyses and compare variability across organoids. However, transcriptome studies focusing on late-stage neural functionality development have been underexplored. Here, we combine and analyze 8 brain organoid transcriptome databases to study the correlation between differentiation protocols and their resulting cellular functionality across various 3D organoid and exogenous brain models. We utilize dimensionality reduction methods including principal component analysis (PCA) and uniform manifold approximation projection (UMAP) to identify and visualize cellular diversity among 3D models and subsequently use gene set enrichment analysis (GSEA) and developmental trajectory inference to quantify neuronal behaviors such as axon guidance, synapse transmission and action potential. We showed high similarity in cellular composition, cellular differentiation pathways and expression of functional genes in human brain organoids during induction and differentiation phases, i.e., up to 3 months in culture. However, during the maturation phase, i.e., 6-month timepoint, we observed significant developmental deficits and depletion of neuronal and astrocytes functional genes as indicated by our GSEA results. Our results caution against use of organoids to model pathophysiology and drug response at this advanced time point and provide insights to tune in vitro iPSC differentiation protocols to achieve desired neuronal functionality and improve current protocols.

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Malankhanova, Tuyana, Lyubov Suldina, Elena Grigor’eva, Sergey Medvedev, Julia Minina, Ksenia Morozova, Elena Kiseleva, Suren Zakian et Anastasia Malakhova. « A Human Induced Pluripotent Stem Cell-Derived Isogenic Model of Huntington’s Disease Based on Neuronal Cells Has Several Relevant Phenotypic Abnormalities ». Journal of Personalized Medicine 10, n^o 4 (9 novembre 2020) : 215. http://dx.doi.org/10.3390/jpm10040215.

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Huntington’s disease (HD) is a severe neurodegenerative disorder caused by a CAG triplet expansion in the first exon of the HTT gene. Here we report the introduction of an HD mutation into the genome of healthy human embryonic fibroblasts through CRISPR/Cas9-mediated homologous recombination. We verified the specificity of the created HTT-editing system and confirmed the absence of undesirable genomic modifications at off-target sites. We showed that both mutant and control isogenic induced pluripotent stem cells (iPSCs) derived by reprogramming of the fibroblast clones can be differentiated into striatal medium spiny neurons. We next demonstrated phenotypic abnormalities in the mutant iPSC-derived neural cells, including impaired neural rosette formation and increased sensitivity to growth factor withdrawal. Moreover, using electron microscopic analysis, we detected a series of ultrastructural defects in the mutant neurons, which did not contain huntingtin aggregates, suggesting that these defects appear early in HD development. Thus, our study describes creation of a new isogenic iPSC-based cell system that models HD and recapitulates HD-specific disturbances in the mutant cells, including some ultrastructural features implemented for the first time.

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Mariani, Alessandro, Davide Comolli, Roberto Fanelli, Gianluigi Forloni et Massimiliano De Paola. « Neonicotinoid Pesticides Affect Developing Neurons in Experimental Mouse Models and in Human Induced Pluripotent Stem Cell (iPSC)-Derived Neural Cultures and Organoids ». Cells 13, n^o 15 (31 juillet 2024) : 1295. http://dx.doi.org/10.3390/cells13151295.

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Neonicotinoids are synthetic, nicotine-derived insecticides used worldwide to protect crops and domestic animals from pest insects. The reported evidence shows that they are also able to interact with mammalian nicotine receptors (nAChRs), triggering detrimental responses in cultured neurons. Exposure to high neonicotinoid levels during the fetal period induces neurotoxicity in animal models. Considering the persistent exposure to these insecticides and the key role of nAChRs in brain development, their potential neurotoxicity on mammal central nervous system (CNS) needs further investigations. We studied here the neurodevelopmental effects of different generations of neonicotinoids on CNS cells in mouse fetal brain and primary cultures and in neuronal cells and organoids obtained from human induced pluripotent stem cells (iPSC). Neonicotinoids significantly affect neuron viability, with imidacloprid (IMI) inducing relevant alterations in synaptic protein expression, neurofilament structures, and microglia activation in vitro, and in the brain of prenatally exposed mouse fetuses. IMI induces neurotoxic effects also on developing human iPSC-derived neurons and cortical organoids. Collectively, the current findings show that neonicotinoids might induce impairment during neuro/immune-development in mouse and human CNS cells and provide new insights in the characterization of risk for the exposure to this class of pesticides.

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Bombieri, Cristina, Andrea Corsi, Elisabetta Trabetti, Alessandra Ruggiero, Giulia Marchetto, Gaetano Vattemi, Maria Teresa Valenti, Donato Zipeto et Maria Grazia Romanelli. « Advanced Cellular Models for Rare Disease Study : Exploring Neural, Muscle and Skeletal Organoids ». International Journal of Molecular Sciences 25, n^o 2 (13 janvier 2024) : 1014. http://dx.doi.org/10.3390/ijms25021014.

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Organoids are self-organized, three-dimensional structures derived from stem cells that can mimic the structure and physiology of human organs. Patient-specific induced pluripotent stem cells (iPSCs) and 3D organoid model systems allow cells to be analyzed in a controlled environment to simulate the characteristics of a given disease by modeling the underlying pathophysiology. The recent development of 3D cell models has offered the scientific community an exceptionally valuable tool in the study of rare diseases, overcoming the limited availability of biological samples and the limitations of animal models. This review provides an overview of iPSC models and genetic engineering techniques used to develop organoids. In particular, some of the models applied to the study of rare neuronal, muscular and skeletal diseases are described. Furthermore, the limitations and potential of developing new therapeutic approaches are discussed.

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Park, Soomin, et Jong-Chan Park. « Advancements in brain organoid models for neurodegenerative disease research ». Organoid 4 (25 décembre 2024) : e12. https://doi.org/10.51335/organoid.2024.4.e12.

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Neurodegenerative diseases (NDs) such as Parkinson’s disease (PD) and Alzheimer’s disease (AD) are progressive disorders characterized by complex, human-specific pathology that poses challenges to drug discovery efforts. Traditional models, including two-dimensional cell cultures and animal models, often fall short in replicating the intricate cellular interactions observed in human neurodegeneration. This review explores the potential of brain organoid technology to address these limitations and offer a model more relevant to humans. Recent advancements in induced pluripotent stem cell (iPSC) technology have enabled the generation of patient-derived brain organoids that differentiate into various neural cell types within 3-dimensional structures. These iPSC-derived brain organoids establish a physiologically relevant microenvironment that mimics human brain architecture and cellular diversity. This review synthesizes studies on the application of brain organoids in modeling PD and AD pathology, including approaches to improve model fidelity. Brain organoids replicate disease-specific features, including dopaminergic neuron degeneration in PD and amyloid plaque formation in AD, offering valuable insights into disease mechanisms and potential therapeutic targets. However, challenges remain, including incomplete maturation, batch variability, and the absence of vascularization and complete cortical layering. Bioengineering approaches, including CRISPR-based gene editing and organ-on-a-chip technologies, are being investigated to overcome these obstacles. Brain organoid technology presents a transformative platform for the study of NDs, facilitating detailed research into disease mechanisms and testing of therapeutics. Overcoming existing challenges is crucial for maximizing the translational value of brain organoids, advancing personalized medicine, and supporting the development of effective therapies for NDs.

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Zhao, Wen-Ning, Chialin Cheng, Kraig M. Theriault, Steven D. Sheridan, Li-Huei Tsai et Stephen J. Haggarty. « A High-Throughput Screen for Wnt/β-Catenin Signaling Pathway Modulators in Human iPSC-Derived Neural Progenitors ». Journal of Biomolecular Screening 17, n^o 9 (24 août 2012) : 1252–63. http://dx.doi.org/10.1177/1087057112456876.

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Wnt/β-catenin signaling has emerged as a central player in pathways implicated in the pathophysiology and treatment of neuropsychiatric disorders. To identify potential novel therapeutics for these disorders, high-throughput screening (HTS) assays reporting on Wnt/β-catenin signaling in disease-relevant contexts are needed. The use of human patient–derived induced pluripotent stem cell (iPSC) models provides ideal disease-relevant context if these stem cell cultures can be adapted for HTS-compatible formats. Here, we describe a sensitive, HTS-compatible Wnt/β-catenin signaling reporter system generated in homogeneous, expandable neural progenitor cells (NPCs) derived from human iPSCs. We validated this system by demonstrating dose-responsive stimulation by several known Wnt/β-catenin signaling pathway modulators, including Wnt3a, a glycogen synthase kinase-3 (GSK3) inhibitor, and the bipolar disorder therapeutic lithium. These responses were robust and reproducible over time across many repeated assays. We then conducted a screen of ~1500 compounds from a library of Food and Drug Administration–approved drugs and known bioactives and confirmed the HTS hits, revealing multiple chemical and biological classes of novel small-molecule probes of Wnt/β-catenin signaling. Generating these type of pathway-selective, cell-based phenotypic assays in human iPSC-derived neural cells will advance the field of human experimental neurobiology toward the goal of identifying and validating targets for neuropsychiatric disorders.

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Tsang, Victoria, Davide Danovi et Ivo Lieberam. « MODL-09. MODELLING MIGRATION OF GLIOBLASTOMA PATIENT-DERIVED CELLS USING HUMAN IPSC-DERIVED NEURAL SPHEROID AND HIGH CONTENT IMAGING ». Neuro-Oncology 24, Supplement_7 (1 novembre 2022) : vii292. http://dx.doi.org/10.1093/neuonc/noac209.1137.

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Abstract Glioblastoma multiforme (GBM) is the most common and aggressive brain tumour in adults. Despite current advances, the existing standard of treatment is ineffective, and the survival prognosis remains 18 months on average from diagnosis. Migrating tumour cells have been implicated in the therapeutic resistance of GBM. They spread by interacting with structures such as white matter tracts and inevitably cause recurrence of the tumour. Valuable cell models able to capture the invasiveness of GBM are critically needed to develop innovative therapies targeting migrating GBM cells. We established an in vitro model mimicking the GBM microenvironment by co-culturing patient-derived GBM cells and human-induced pluripotent stem cell-derived cortical neural spheroids with radiating axons. Using high content imaging, we developed a robust workflow to quantify the GBM cells' infiltration of the neural spheroid in endpoint assays. Images were acquired on the Operetta CLS and analysed using Harmony Imaging and Analysis Software. We also performed live imaging assays using the Phasefocus Livecyte, in which we studied the directionality, displacement and speed of the GBM cells engaged on axons. Our data indicate that GBM cells change morphology when cultured on axons and that they migrated towards the neural spheroid once engaged on axons. We showed that patient derived-cell lines representing different subtypes vary in migratory properties as well as in levels of infiltration capability. Finally, we used this model to test antagonists inhibiting pathways involved in the migration of GBM cells. To summarise, we developed a novel model able to mimic the GBM migration on axons and able to screen for compounds affecting cell migration which could potentially offer innovative precision-medicine therapies.

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Galera-Monge, Teresa, Francisco Zurita-Díaz, Isaac Canals, Marita Grønning Hansen, Laura Rufián-Vázquez, Johannes K. Ehinger, Eskil Elmér et al. « Mitochondrial Dysfunction and Calcium Dysregulation in Leigh Syndrome Induced Pluripotent Stem Cell Derived Neurons ». International Journal of Molecular Sciences 21, n^o 9 (30 avril 2020) : 3191. http://dx.doi.org/10.3390/ijms21093191.

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Leigh syndrome (LS) is the most frequent infantile mitochondrial disorder (MD) and is characterized by neurodegeneration and astrogliosis in the basal ganglia or the brain stem. At present, there is no cure or treatment for this disease, partly due to scarcity of LS models. Current models generally fail to recapitulate important traits of the disease. Therefore, there is an urgent need to develop new human in vitro models. Establishment of induced pluripotent stem cells (iPSCs) followed by differentiation into neurons is a powerful tool to obtain an in vitro model for LS. Here, we describe the generation and characterization of iPSCs, neural stem cells (NSCs) and iPSC-derived neurons harboring the mtDNA mutation m.13513G>A in heteroplasmy. We have performed mitochondrial characterization, analysis of electrophysiological properties and calcium imaging of LS neurons. Here, we show a clearly compromised oxidative phosphorylation (OXPHOS) function in LS patient neurons. This is also the first report of electrophysiological studies performed on iPSC-derived neurons harboring an mtDNA mutation, which revealed that, in spite of having identical electrical properties, diseased neurons manifested mitochondrial dysfunction together with a diminished calcium buffering capacity. This could lead to an overload of cytoplasmic calcium concentration and the consequent cell death observed in patients. Importantly, our results highlight the importance of calcium homeostasis in LS pathology.

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Castellanos-Montiel, María José, Mathilde Chaineau, Anna Kristyna Franco-Flores, Ghazal Haghi, Dulce Carrillo-Valenzuela, Wolfgang E. Reintsch, Carol X. Q. Chen et Thomas M. Durcan. « An Optimized Workflow to Generate and Characterize iPSC-Derived Motor Neuron (MN) Spheroids ». Cells 12, n^o 4 (8 février 2023) : 545. http://dx.doi.org/10.3390/cells12040545.

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A multitude of in vitro models based on induced pluripotent stem cell (iPSC)-derived motor neurons (MNs) have been developed to investigate the underlying causes of selective MN degeneration in motor neuron diseases (MNDs). For instance, spheroids are simple 3D models that have the potential to be generated in large numbers that can be used across different assays. In this study, we generated MN spheroids and developed a workflow to analyze them. To start, the morphological profiling of the spheroids was achieved by developing a pipeline to obtain measurements of their size and shape. Next, we confirmed the expression of different MN markers at the transcript and protein levels by qPCR and immunocytochemistry of tissue-cleared samples, respectively. Finally, we assessed the capacity of the MN spheroids to display functional activity in the form of action potentials and bursts using a microelectrode array approach. Although most of the cells displayed an MN identity, we also characterized the presence of other cell types, namely interneurons and oligodendrocytes, which share the same neural progenitor pool with MNs. In summary, we successfully developed an MN 3D model, and we optimized a workflow that can be applied to perform its morphological, gene expression, protein, and functional profiling over time.

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Tsang, V. S., I. Lieberam et D. Danovi. « P17.03.B Modelling migration of glioblastoma patient-derived cells using human iPSC-derived neural spheroid and high content analysis ». Neuro-Oncology 24, Supplement_2 (1 septembre 2022) : ii89. http://dx.doi.org/10.1093/neuonc/noac174.311.

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Abstract Background Glioblastoma multiforme (GBM) is the most common and aggressive brain tumour in adults. Despite current advances, the existing standard of treatment is ineffective, and the survival prognosis remains just over a year from diagnosis. Migrating tumour cells have been implicated in the therapeutic resistance of GBM. They spread by interacting with structures such as white matter tracts and inevitably cause recurrence of the tumour. Valuable cell models able to capture the invasiveness of GBM are critically needed to develop innovative therapies targeting migrating GBM cells. Material and Methods We established an in vitro model mimicking the GBM microenvironment by co-culturing patient-derived GBM cells and human induced pluripotent stem cell-derived cortical neural spheroids with radiating axons. Using high content imaging, we developed a robust workflow to quantify the GBM cells infiltration of the neural spheroid in endpoint assays. Images were acquired on the Operetta CLS high content device (Perkin Elmer) and analysed using the built-in Harmony Imaging and Analysis Software. We also performed live imaging assays using the Livecyte quantitative phase imager (Phasefocus), in which we studied the directionality, displacement and speed of the GBM cells engaged on axons. Results Our data indicate that GBM cells change morphology when cultured on axons and that they migrated towards the neural spheroid once engaged on axons. We showed that cell lines from different patients vary in migratory properties as well as in levels of infiltration capability. Finally, we used this model to test several antagonists to pathways involved in migration of GBM cells. Conclusion The main deliverable of this project is the setup of a novel model able to mimic the GBM migration on axons and able to screen for compounds affecting cell migration. This could potentially offer innovative precision-medicine therapies.

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Nayak, Ritu, Idan Rosh, Irina Kustanovich et Shani Stern. « Mood Stabilizers in Psychiatric Disorders and Mechanisms Learnt from In Vitro Model Systems ». International Journal of Molecular Sciences 22, n^o 17 (27 août 2021) : 9315. http://dx.doi.org/10.3390/ijms22179315.

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Bipolar disorder (BD) and schizophrenia are psychiatric disorders that manifest unusual mental, behavioral, and emotional patterns leading to suffering and disability. These disorders span heterogeneous conditions with variable heredity and elusive pathophysiology. Mood stabilizers such as lithium and valproic acid (VPA) have been shown to be effective in BD and, to some extent in schizophrenia. This review highlights the efficacy of lithium and VPA treatment in several randomized, controlled human trials conducted in patients suffering from BD and schizophrenia. Furthermore, we also address the importance of using induced pluripotent stem cells (iPSCs) as a disease model for mirroring the disease’s phenotypes. In BD, iPSC-derived neurons enabled finding an endophenotype of hyperexcitability with increased hyperpolarizations. Some of the disease phenotypes were significantly alleviated by lithium treatment. VPA studies have also reported rescuing the Wnt/β-catenin pathway and reducing activity. Another significant contribution of iPSC models can be attributed to studying the molecular etiologies of schizophrenia such as abnormal differentiation of patient-derived neural stem cells, decreased neuronal connectivity and neurite number, impaired synaptic function, and altered gene expression patterns. Overall, despite significant advances using these novel models, much more work remains to fully understand the mechanisms by which these disorders affect the patients’ brains.

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Mansur, Fernanda, André Luiz Teles e Silva, Ana Karolyne Santos Gomes, Juliana Magdalon, Janaina Sena de Souza, Karina Griesi-Oliveira, Maria Rita Passos-Bueno et Andréa Laurato Sertié. « Complement C4 Is Reduced in iPSC-Derived Astrocytes of Autism Spectrum Disorder Subjects ». International Journal of Molecular Sciences 22, n^o 14 (15 juillet 2021) : 7579. http://dx.doi.org/10.3390/ijms22147579.

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In recent years, accumulating evidence has shown that the innate immune complement system is involved in several aspects of normal brain development and in neurodevelopmental disorders, including autism spectrum disorder (ASD). Although abnormal expression of complement components was observed in post-mortem brain samples from individuals with ASD, little is known about the expression patterns of complement molecules in distinct cell types in the developing autistic brain. In the present study, we characterized the mRNA and protein expression profiles of a wide range of complement system components, receptors and regulators in induced pluripotent stem cell (iPSC)-derived neural progenitor cells, neurons and astrocytes of individuals with ASD and neurotypical controls, which constitute in vitro cellular models that recapitulate certain features of both human brain development and ASD pathophysiology. We observed that all the analyzed cell lines constitutively express several key complement molecules. Interestingly, using different quantification strategies, we found that complement C4 mRNA and protein are expressed in significantly lower levels by astrocytes derived from ASD individuals compared to control astrocytes. As astrocytes participate in synapse elimination, and diminished C4 levels have been linked to defective synaptic pruning, our findings may contribute to an increased understanding of the atypically enhanced brain connectivity in ASD.

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Maussion, Gilles, Cecilia Rocha, Narges Abdian, Dimitri Yang, Julien Turk, Dulce Carrillo Valenzuela, Luisa Pimentel et al. « Transcriptional Dysregulation and Impaired Neuronal Activity in FMR1 Knock-Out and Fragile X Patients’ iPSC-Derived Models ». International Journal of Molecular Sciences 24, n^o 19 (5 octobre 2023) : 14926. http://dx.doi.org/10.3390/ijms241914926.

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Fragile X syndrome (FXS) is caused by a repression of the FMR1 gene that codes the Fragile X mental retardation protein (FMRP), an RNA binding protein involved in processes that are crucial for proper brain development. To better understand the consequences of the absence of FMRP, we analyzed gene expression profiles and activities of cortical neural progenitor cells (NPCs) and neurons obtained from FXS patients’ induced pluripotent stem cells (IPSCs) and IPSC-derived cells from FMR1 knock-out engineered using CRISPR-CAS9 technology. Multielectrode array recordings revealed in FMR1 KO and FXS patient cells, decreased mean firing rates; activities blocked by tetrodotoxin application. Increased expression of presynaptic mRNA and transcription factors involved in the forebrain specification and decreased levels of mRNA coding AMPA and NMDA subunits were observed using RNA sequencing on FMR1 KO neurons and validated using quantitative PCR in both models. Intriguingly, 40% of the differentially expressed genes were commonly deregulated between NPCs and differentiating neurons with significant enrichments in FMRP targets and autism-related genes found amongst downregulated genes. Our findings suggest that the absence of FMRP affects transcriptional profiles since the NPC stage, and leads to impaired activity and neuronal differentiation over time, which illustrates the critical role of FMRP protein in neuronal development.

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Costamagna, Gianluca, Luca Andreoli, Stefania Corti et Irene Faravelli. « iPSCs-Based Neural 3D Systems : A Multidimensional Approach for Disease Modeling and Drug Discovery ». Cells 8, n^o 11 (14 novembre 2019) : 1438. http://dx.doi.org/10.3390/cells8111438.

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Induced pluripotent stem cells (iPSCs)-based two-dimensional (2D) protocols have offered invaluable insights into the pathophysiology of neurological diseases. However, these systems are unable to reproduce complex cytoarchitectural features, cell-cell and tissue-tissue interactions like their in vivo counterpart. Three-dimensional (3D)-based culture protocols, though in their infancy, have offered new insights into modeling human diseases. Human neural organoids try to recapitulate the cellular diversity of complex tissues and can be generated from iPSCs to model the pathophysiology of a wide spectrum of pathologies. The engraftment of iPSCs into mice models and the improvement of differentiation protocols towards 3D cultures has enabled the generation of more complex multicellular systems. Consequently, models of neuropsychiatric disorders, infectious diseases, brain cancer and cerebral hypoxic injury can now be investigated from new perspectives. In this review, we consider the advancements made in modeling neuropsychiatric and neurological diseases with iPSC-derived organoids and their potential use to develop new drugs.

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Ng, Neville S., Simon Maksour, Jeremy S. Lum, Michelle Newbery, Victoria Shephard et Lezanne Ooi. « An Optimized Direct Lysis Gene Expression Microplate Assay and Applications for Disease, Differentiation, and Pharmacological Cell-Based Studies ». Biosensors 12, n^o 6 (26 mai 2022) : 364. http://dx.doi.org/10.3390/bios12060364.

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Routine cell culture reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) gene expression analysis is limited in scalability due to minimum sample requirement and multistep isolation procedures. In this study, we aimed to optimize and apply a cost-effective and rapid protocol for directly sampling gene expression data from microplate cell cultures. The optimized protocol involves direct lysis of microplate well population followed by a reduced thermocycler reaction time one-step RT-qPCR assay. In applications for inflammation and stress-induced cell-based models, the direct lysis RT-qPCR microplate assay was utilized to detect IFN1 and PPP1R15A expression by poly(I:C) treated primary fibroblast cultures, IL6 expression by poly(I:C) iPSC-derived astrocytes, and differential PPP1R15A expression by ER-stressed vanishing white-matter disease patient induced pluripotent stem cell (iPSC)-derived astrocytes. Neural differentiation recipe optimizations were performed with SYN1 and VGLUT1 expression in neuronal cultures, and S100B, GFAP, and EAAT1 expression in astrocyte differentiations. The protocol provides microplate gene expression results from cell lysate to readout within ~35 min, with comparable cost to routine RT-qPCR, and it may be utilized to support laboratory cell-based assays in basic and applied scientific and medical fields of research including stem-cell differentiation, cell physiology, and drug mechanism studies.

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Buijsen, Ronald A. M., Linda M. van der Graaf, Elsa C. Kuijper, Barry A. Pepers, Elena Daoutsali, Lotte Weel, Vered Raz, David A. Parfitt et Willeke M. C. van Roon-Mom. « Calcium-Enhanced Medium-Based Delivery of Splice Modulating Antisense Oligonucleotides in 2D and 3D hiPSC-Derived Neuronal Models ». Biomedicines 12, n^o 9 (23 août 2024) : 1933. http://dx.doi.org/10.3390/biomedicines12091933.

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Antisense technology demonstrates significant potential for addressing inherited brain diseases, with over a dozen products already available and numerous others in the development pipeline. The versatility of differentiating induced pluripotent stem cells (iPSCs) into nearly all neural cell types proves invaluable for comprehending the mechanisms behind neurological diseases, replicating cellular phenotypes, and advancing the testing and development of new therapies, including antisense oligonucleotide therapeutics. While delivering antisense oligonucleotides (ASOs) to human iPSC-based neuronal models has posed challenges, this study explores various delivery methods, including lipid-based transfection, gymnotic uptake, Ca(2+)-enhanced medium (CEM)-based delivery, and electroporation, in 2D and 3D hiPSC-derived neuronal models. This study reveals that CEM-based delivery exhibits efficiency and low toxicity in both 2D neuronal cultures and 3D brain organoids. Furthermore, the findings indicate that CEM is slightly more effective in neurons than in astrocytes, suggesting promising avenues for further exploration and optimization of preclinical ASO strategies in the treatment of neurological disorders.

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Deshmukh, Rahul S., Krisztián A. Kovács et András Dinnyés. « Drug Discovery Models and Toxicity Testing Using Embryonic and Induced Pluripotent Stem-Cell-Derived Cardiac and Neuronal Cells ». Stem Cells International 2012 (2012) : 1–9. http://dx.doi.org/10.1155/2012/379569.

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Development of induced pluripotent stem cells (iPSCs) using forced expression of specific sets of transcription factors has changed the field of stem cell research extensively. Two important limitations for research application of embryonic stem cells (ESCs), namely, ethical and immunological issues, can be circumvented using iPSCs. Since the development of first iPSCs, tremendous effort has been directed to the development of methods to increase the efficiency of the process and to reduce the extent of genomic modifications associated with the reprogramming procedure. The established lineage-specific differentiation protocols developed for ESCs are being applied to iPSCs, as they have great potential in regenerative medicine for cell therapy, disease modeling either for drug development or for fundamental science, and, last but not least, toxicity testing. This paper reviews efforts aimed at practical development of iPSC differentiation to neural/cardiac lineages and further the use of these iPSCs-derived cells for drug development and toxicity testing.

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Song, Liqing, Yuanwei Yan, Mark Marzano et Yan Li. « Studying Heterotypic Cell–Cell Interactions in the Human Brain Using Pluripotent Stem Cell Models for Neurodegeneration ». Cells 8, n^o 4 (1 avril 2019) : 299. http://dx.doi.org/10.3390/cells8040299.

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Human cerebral organoids derived from induced pluripotent stem cells (iPSCs) provide novel tools for recapitulating the cytoarchitecture of the human brain and for studying biological mechanisms of neurological disorders. However, the heterotypic interactions of neurovascular units, composed of neurons, pericytes (i.e., the tissue resident mesenchymal stromal cells), astrocytes, and brain microvascular endothelial cells, in brain-like tissues are less investigated. In addition, most cortical organoids lack a microglia component, the resident immune cells in the brain. Impairment of the blood-brain barrier caused by improper crosstalk between neural cells and vascular cells is associated with many neurodegenerative disorders. Mesenchymal stem cells (MSCs), with a phenotype overlapping with pericytes, have promotion effects on neurogenesis and angiogenesis, which are mainly attributed to secreted growth factors and extracellular matrices. As the innate macrophages of the central nervous system, microglia regulate neuronal activities and promote neuronal differentiation by secreting neurotrophic factors and pro-/anti-inflammatory molecules. Neuronal-microglia interactions mediated by chemokines signaling can be modulated in vitro for recapitulating microglial activities during neurodegenerative disease progression. In this review, we discussed the cellular interactions and the physiological roles of neural cells with other cell types including endothelial cells and microglia based on iPSC models. The therapeutic roles of MSCs in treating neural degeneration and pathological roles of microglia in neurodegenerative disease progression were also discussed.

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Stebbins, Matthew J., Benjamin D. Gastfriend, Scott G. Canfield, Ming-Song Lee, Drew Richards, Madeline G. Faubion, Wan-Ju Li, Richard Daneman, Sean P. Palecek et Eric V. Shusta. « Human pluripotent stem cell–derived brain pericyte–like cells induce blood-brain barrier properties ». Science Advances 5, n^o 3 (mars 2019) : eaau7375. http://dx.doi.org/10.1126/sciadv.aau7375.

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Brain pericytes play important roles in the formation and maintenance of the neurovascular unit (NVU), and their dysfunction has been implicated in central nervous system disorders. While human pluripotent stem cells (hPSCs) have been used to model other NVU cell types, including brain microvascular endothelial cells (BMECs), astrocytes, and neurons, hPSC-derived brain pericyte–like cells have not been integrated into these models. In this study, we generated neural crest stem cells (NCSCs), the embryonic precursor to forebrain pericytes, from hPSCs and subsequently differentiated NCSCs to brain pericyte–like cells. These cells closely resembled primary human brain pericytes and self-assembled with endothelial cells. The brain pericyte–like cells induced blood-brain barrier properties in BMECs, including barrier enhancement and reduced transcytosis. Last, brain pericyte–like cells were incorporated with iPSC-derived BMECs, astrocytes, and neurons to form an isogenic human model that should prove useful for the study of the NVU.

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Skinner, Kasey, Tomoyuki Koga, Shunichiro Miki, Robert F. Gruener, R. Stephanie Huang, Frank Furnari et C. Ryan Miller. « HGG-12. HUMAN IPSC-DERIVED H3.3K27M NEUROSPHERES : A NOVEL MODEL FOR INVESTIGATING DIPG PATHOGENESIS AND DRUG RESPONSE ». Neuro-Oncology 23, Supplement_1 (1 juin 2021) : i19—i20. http://dx.doi.org/10.1093/neuonc/noab090.078.

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Abstract Diffuse intrinsic pontine glioma (DIPG) is a subset of high-grade glioma that occurs predominantly in children and has no cure. Up to 80% of DIPG harbor a heterozygous point mutation that results in a lysine 27 to methionine substitution in histone variant H3.3 (H3.3K27M). Existing DIPG models have provided insight into the role of H3.3K27M but have limitations: genetically engineered murine models often rely on overexpression of the mutant histone to form tumors; patient-derived xenografts (PDX) are more genetically faithful but preclude examination of the effect of individual mutations on pathogenesis. To address these shortcomings and better recapitulate the genetics of human tumors, we designed a novel DIPG model based on human induced pluripotent stem cells (iPSC) edited via CRISPR to express heterozygous H3.3K27M. Edited iPSC were chemically differentiated into neural progenitor cells, which upon implantation into the brainstems of immunodeficient mice formed diffusely invasive tumors that were histologically consistent with high-grade glioma. Further, neurospheres cultured from primary tumors formed secondary tumors upon reimplantation with more diffuse invasion, suggesting in vivo evolution. To validate this model’s relevance to DIPG transcriptionally, we performed RNA-sequencing on a cohort of primary and secondary tumor neurospheres (termed primary and secondary iDIPG) and compared them to published RNA-seq data from pediatric PDX and patient tumor samples. Hierarchical clustering and principal component analysis on differentially expressed genes (P<0.05) showed that H3.3K27M iDIPG cluster with H3.3K27M PDX and patient tumors. Further, ssGSEA showed that H3.3K27M iDIPG are enriched for astrocytic and mesenchymal signature genes, a defining feature of H3.3K27M DIPG. Finally, we found that primary H3.3K27M iDIPG neurospheres are sensitive to panobinostat, an HDAC inhibitor shown to be effective against H3.3K27M DIPG cells in vitro. Overall, these data suggest that H3.3K27M iDIPG are a promising tool for investigating DIPG biology and new therapeutic strategies.

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Manz, Frederik, Daniel Haag, Stefan M. Pfister et Lena Kutscher. « MEDB-22. iPSC-derived cerebellar organoid model for hereditary genetic predisposition in SHH-medulloblastoma ». Neuro-Oncology 24, Supplement_1 (1 juin 2022) : i109. http://dx.doi.org/10.1093/neuonc/noac079.396.

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Abstract Medulloblastoma is one of the most common malignant embryonal brain tumors in children. Medulloblastomas of the Sonic Hedgehog (SHH) group arise from excessive proliferation of granule neuron progenitor (GNP) cells during cerebellar development. Genetic predisposition accounts for nearly 40% of all pediatric SHH-medulloblastomas. Recently, ELP1, a novel predisposition gene, was shown to be germline mutated in 15% of SHH-medulloblastoma patients. ELP1 encodes the scaffolding member of the Elongator complex and is required for efficient translation. Heterozygous mutations in ELP1 have been associated with the neural disorder Familial Dysautonomia, but not cancer. ELP1-associated medulloblastomas frequently harbor somatic PTCH1 co-mutations. It remains unclear how ELP1 affects the GNP lineage during normal cerebellar development and tumorigenesis in pediatric SHH-medulloblastoma patients. To characterize ELP1 mutations in the GNP lineage in vitro, we established a cerebellar organoid model from human induced pluripotent stem cells (iPSCs). We genetically inserted an EGFP reporter downstream of the endogenous GNP-specific ATOH1 locus in control iPSCs and generated cerebellar organoids according to published protocols. Marker gene and protein expression levels confirmed the cerebellar identity of the 3D model. Furthermore, activation of the EGFP reporter in single cells within the organoid highlighted the specification of putative GNPs. Next, we will determine the specific cell state of putative iGNPs and compare to human GNPs identified in our scRNAseq cerebellum atlas. To analyze tumorigenesis through ELP1 loss, we will introduce patient-specific ELP1mutations into ATOH1-EGFP iPSCs. Cerebellar organoids derived from ELP1-, PTCH1-deficient and control iPSCs will serve as models to study GNP proliferation, differentiation, apoptosis and tumor formation. Combining genome editing, in vitro 3D differentiation and functional studies, we will characterize the novel predisposition gene ELP1in GNPs during cerebellar development. In addition, we will determine the interplay of ELP1and PTCH1 co-mutations, predisposing SHH-medulloblastoma formation.

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Csobonyeiova, Maria, Stefan Polak et Lubos Danisovic. « Recent Overview of the Use of iPSCs Huntington’s Disease Modeling and Therapy ». International Journal of Molecular Sciences 21, n^o 6 (24 mars 2020) : 2239. http://dx.doi.org/10.3390/ijms21062239.

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Huntington’s disease (HD) is an inherited, autosomal dominant, degenerative disease characterized by involuntary movements, cognitive decline, and behavioral impairment ending in death. HD is caused by an expansion in the number of CAG repeats in the huntingtin gene on chromosome 4. To date, no effective therapy for preventing the onset or progression of the disease has been found, and many symptoms do not respond to pharmacologic treatment. However, recent results of pre-clinical trials suggest a beneficial effect of stem-cell-based therapy. Induced pluripotent stem cells (iPSCs) represent an unlimited cell source and are the most suitable among the various types of autologous stem cells due to their patient specificity and ability to differentiate into a variety of cell types both in vitro and in vivo. Furthermore, the cultivation of iPSC-derived neural cells offers the possibility of studying the etiopathology of neurodegenerative diseases, such as HD. Moreover, differentiated neural cells can organize into three-dimensional (3D) organoids, mimicking the complex architecture of the brain. In this article, we present a comprehensive review of recent HD models, the methods for differentiating HD–iPSCs into the desired neural cell types, and the progress in gene editing techniques leading toward stem-cell-based therapy.

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McIntyre, Laura Lynn, Warren Plaisted, Ronald Coleman, Jeanne Loring, Thomas Lane et Craig M. Walsh. « Evaluating the Therapeutic Potential of Transplantation of Neural Precursor Cells for Treating the Autoimmune Disease Multiple Sclerosis ». Journal of Immunology 198, n^o 1_Supplement (1 mai 2017) : 219.1. http://dx.doi.org/10.4049/jimmunol.198.supp.219.1.

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Abstract Multiple Sclerosis is a chronic, autoimmune disease for which there is currently no cure. Previous studies suggest transplantation of neural precursor cells (NPCs) is a promising therapeutic strategy to treat neurological disorders. Using two mouse models of MS, experimental autoimmune encephalomyelitis (EAE) and murine hepatitis virus (MHV), we have observed remyelination and decreased neuroinflammation when mice received an intra-spinal transplant of syngeneic mouse GFP-NPCs. This recovery was due to replacement of damaged cells and had little effect upon immune responses. In contrast, transplantation of xenogeneic human embryonic stem cell (hESC) derived NPCs (hNPCs) or induced pluripotent stem cell (iPSC) derived NPCs (hiNPCs) modulates the host immune response. Mice receiving intra-spinal transplants of human NPCs displayed less demyelination, decreased neuroinflammation, and an increase in T regulatory cells (Tregs). Recovery was not a result of cell replacement by hNPCs because transplanted cells underwent rapid xenograft rejection. Importantly, ablation of Tregs abrogated clinical and histopathological improvement mediated by hNPC transplantation. Furthermore, hNPCs promote the induction of naïve T cells into Tregs in co-cultures in vitro. However, the mechanisms and factors that propagate Treg induction and function resulting in subsequent endogenous remyelination remain undiscovered. We hypothesize rejection of NPCs plays a role in generation of Tregs in vitro and in vivo. Future experiments will focus on elucidating the factors from hNPCs that are responsible for Treg differentiation and evaluate the ability of syngeneic, allogeneic, and xenogeneic NPCs to generate Tregs.

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Nizzardo, Monica, Monica Bucchia, Agnese Ramirez, Elena Trombetta, Nereo Bresolin, Giacomo P. Comi et Stefania Corti. « iPSC-derived LewisX+CXCR4+β1-integrin+ neural stem cells improve the amyotrophic lateral sclerosis phenotype by preserving motor neurons and muscle innervation in human and rodent models ». Human Molecular Genetics 25, n^o 15 (6 juin 2016) : 3152–63. http://dx.doi.org/10.1093/hmg/ddw163.

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Johnston, *Jenessa, Brandi Quintanilla, Peixiong Yuan, Shiyong Peng, Mani Yavi, Hector Caruncho, Bashkim Kadriu et Carlos Zarate Jr. « DIFFERENCE IN TREATMENT-RESPONSE OF IPSC-DERIVED NEURONS TO REELIN AND (2R,6R)-HNK FROM PARTICIPANTS WITH TREATMENT- RESISTANT DEPRESSION ». International Journal of Neuropsychopharmacology 28, Supplement_1 (février 2025) : i327. https://doi.org/10.1093/ijnp/pyae059.583.

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Abstract Background Treatment-resistant depression (TRD) presents a significant clinical challenge, with around 30% of patients failing to respond to first-line treatments. Ketamine, administered at subanesthetic doses, has demonstrated rapid and robust antidepressant effects in TRD patients. Some research has suggested that ketamine's fast-acting antidepressant effects are linked to the transient activation of the mammalian target of rapamycin (mTOR), which enhances synaptic signaling by upregulating proteins such GluA1. A major ketamine metabolite, (2R,6R)-hydroxynorketamine (HNK), appears to produce similar effects without the adverse side effects or N-methyl-d-asparate receptor inhibition. Additionally, reelin, a glycoprotein crucial for proper cortical development, seems to mirror some of the fast-acting antidepressant-like effects of ketamine in pre-clinical models, warranting further exploration. Aims & Objectives Characterize neurons derived from induced pluripotent stem cells (iPSCs) collected from participants with TRD. Baseline differences in protein expression were analyzed, as well as varying treatment-response to (2R,6R)-HNK and reelin. Method iPSCs were generated from peripheral mononuclear blood cells collected from individuals with TRD (n=5) and healthy controls (HCs) (n=6). The STEMdiffTM embryoid body protocol was followed for 19 days to develop single-cell neural progenitor cells. After 10 weeks of cultivation, cells were exposed to five different conditions (vehicle + DMSO, 5nM reelin, 10nM reelin, 50nM reelin, 1μ M (2R,6R)-HNK) at two time points (1 hour and 24 hours) to assess short- and long-term effects of reelin and (2R,6R)-HNK. Western blotting analyses were employed to measure mTOR, phosphorylated-mTOR (p-mTOR), post-synaptic density 95 (PSD95), GluA1, Synapsin 1 (Syn-I), Disabled-1 (Dab1), tyrosine kinase receptor B (TrkB), extracellular signal-regulated kinase (ERK), and p-ERK. Results In TRD cell lines at 1-hour, both (2R,6R)-HNK and reelin significantly increased levels of PSD-95 (50nM reelin, p<0.05; HNK 1μ M, p<0.01), Syn-I (10nM reelin, p<0.05; 50nM reelin, p<0.01; HNK 1μ M, p<0.01), Dab1 (10nM reelin, p<0.05; 50nM reelin, p<0.01; HNK 1μ M, p<0.01), and p-ERK (50nM reelin, p<0.05; HNK 1μ M, p<0.05). After 24 hours, these effects were reversed, showing significant down- regulation of PSD-95 (10nM reelin, p<0.05; 50nM reelin, p<0.01) and Syn-I (10nM reelin, p<0.05; 50nM reelin, p<0.01; HNK 1μ M, p<0.01). However, no significant differences were found in any treatment or time group in healthy controls. Discussion & Conclusion The high rate of treatment failure in TRD necessitates the exploration of novel antidepressants with reduced long-term side effects, higher efficacy, and rapid therapeutic action. This preliminary study indicates that both reelin and (2R,6R)-HNK show promise in activating cellular pathways crucial for fast-acting antidepressant responses. The disparities observed between TRD and HC cell lines warrant further investigation. Additionally, the translation of novel antidepressants into clinical practice has been hindered by the lack of in vitro or in vivo models capable of representing the heterogeneity of depression. While still in early stages, iPSCs could offer a more precise proxy for evaluating treatment responsiveness, contributing to the development of innovative antidepressant therapeutics and improved translation in pharmacological research.

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Frawley, Lauren, Noam Tomer Taylor, Olivia Sivills, Ella McPhillamy, Timothy Duy To, Yibo Wu, Beek Yoke Chin et Chiew Yen Wong. « Stem Cell Therapy for the Treatment of Amyotrophic Lateral Sclerosis : Comparison of the Efficacy of Mesenchymal Stem Cells, Neural Stem Cells, and Induced Pluripotent Stem Cells ». Biomedicines 13, n^o 1 (27 décembre 2024) : 35. https://doi.org/10.3390/biomedicines13010035.

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Background/Objectives: Amyotrophic lateral sclerosis (ALS), or Lou Gehrig’s disease, is a debilitating, incurable neurodegenerative disorder characterised by motor neuron death in the spinal cord, brainstem, and motor cortex. With an incidence rate of about 4.42 cases per 100,000 people annually, ALS severely impacts motor function and quality of life, causing progressive muscle atrophy, spasticity, paralysis, and eventually death. The cause of ALS is largely unknown, with 90% of cases being sporadic and 10% familial. Current research targets molecular mechanisms of inflammation, excitotoxicity, aggregation-prone proteins, and proteinopathy. Methods: This review evaluates the efficacy of three stem cell types in ALS treatment: mesenchymal stem cells (MSCs), neural stem cells (NSCs), and induced pluripotent stem cells (iPSCs). Results: MSCs, derived from various tissues, show neuroprotective and regenerative qualities, with clinical trials suggesting potential benefits but limited by small sample sizes and non-randomised designs. NSCs, isolated from the fetal spinal cord or brain, demonstrate promise in animal models but face functional integration and ethical challenges. iPSCs, created by reprogramming patient-specific somatic cells, offer a novel approach by potentially replacing or supporting neurons. iPSC therapy addresses ethical issues related to embryonic stem cells but encounters challenges regarding genotoxicity and epigenetic irregularities, somatic cell sources, privacy concerns, the need for extensive clinical trials, and high reprogramming costs. Conclusions: This research is significant for advancing ALS treatment beyond symptomatic relief and modest survival extensions to actively modifying disease progression and improving patient outcomes. Successful stem cell therapies could lead to new ALS treatments, slowing motor function loss and reducing symptom severity.

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De Beuckeleer, Sarah, Tim Van De Looverbosch, Johanna Van Den Daele, Peter Ponsaerts et Winnok H. De Vos. « Unbiased identification of cell identity in dense mixed neural cultures ». eLife 13 (17 janvier 2025). https://doi.org/10.7554/elife.95273.4.

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Induced pluripotent stem cell (iPSC) technology is revolutionizing cell biology. However, the variability between individual iPSC lines and the lack of efficient technology to comprehensively characterize iPSC-derived cell types hinder its adoption in routine preclinical screening settings. To facilitate the validation of iPSC-derived cell culture composition, we have implemented an imaging assay based on cell painting and convolutional neural networks to recognize cell types in dense and mixed cultures with high fidelity. We have benchmarked our approach using pure and mixed cultures of neuroblastoma and astrocytoma cell lines and attained a classification accuracy above 96%. Through iterative data erosion, we found that inputs containing the nuclear region of interest and its close environment, allow achieving equally high classification accuracy as inputs containing the whole cell for semi-confluent cultures and preserved prediction accuracy even in very dense cultures. We then applied this regionally restricted cell profiling approach to evaluate the differentiation status of iPSC-derived neural cultures, by determining the ratio of postmitotic neurons and neural progenitors. We found that the cell-based prediction significantly outperformed an approach in which the population-level time in culture was used as a classification criterion (96% vs 86%, respectively). In mixed iPSC-derived neuronal cultures, microglia could be unequivocally discriminated from neurons, regardless of their reactivity state, and a tiered strategy allowed for further distinguishing activated from non-activated cell states, albeit with lower accuracy. Thus, morphological single-cell profiling provides a means to quantify cell composition in complex mixed neural cultures and holds promise for use in the quality control of iPSC-derived cell culture models.

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Sarieva, Kseniia, Felix Hildebrand, Theresa Kagermeier, Zeynep Yentür, Katharina Becker et Simone Mayer. « Pluripotent stem cell-derived neural progenitor cells can be used to model effects of IL-6 on human neurodevelopment ». Disease Models & ; Mechanisms 16, n^o 11 (1 novembre 2023). http://dx.doi.org/10.1242/dmm.050306.

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ABSTRACT Maternal immune activation (MIA) increases the risks for neurodevelopmental disorders in offspring through inflammatory cytokines, including interleukin-6 (IL-6). We therefore aimed to establish a human two-dimensional (2D) in vitro neural model to investigate the effects of IL-6 exposure on neurodevelopment. IL-6 signal transduction requires two receptors: interleukin-6 signal transducer (IL6ST) and interleukin-6 receptor (IL6R). Prenatally, neural cells lack IL6R, and hence cannot elicit cis IL-6 signaling, but IL6R can be provided by microglia in trans. We demonstrate here that an immortalized human neural progenitor cell (NPC) line, ReNCell CX, expresses IL6ST and elicits both cis and trans IL-6 signaling, limiting its use as a model of MIA. In contrast, induced pluripotent stem cell (iPSC)-derived NPCs only activate the IL-6 cascade in trans. Activation of the trans IL-6 cascade did not result in increased proliferation of iPSC-derived NPCs or ReNCell CX, as has been demonstrated in animal models. iPSC-derived NPCs upregulated NR2F1 expression in response to IL-6 signaling in line with analogous experiments in organoids. Thus, iPSC-derived NPCs can be used to model gene expression changes in response to MIA in 2D cultures.

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Tomov, Martin L., Alison O’Neil, Hamdah S. Abbasi, Beth A. Cimini, Anne E. Carpenter, Lee L. Rubin et Mark Bathe. « Resolving cell state in iPSC-derived human neural samples with multiplexed fluorescence imaging ». Communications Biology 4, n^o 1 (24 juin 2021). http://dx.doi.org/10.1038/s42003-021-02276-x.

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AbstractHuman induced pluripotent stem cell-derived (iPSC) neural cultures offer clinically relevant models of human diseases, including Amyotrophic Lateral Sclerosis, Alzheimer’s, and Autism Spectrum Disorder. In situ characterization of the spatial-temporal evolution of cell state in 3D culture and subsequent 2D dissociated culture models based on protein expression levels and localizations is essential to understanding neural cell differentiation, disease state phenotypes, and sample-to-sample variability. Here, we apply PRobe-based Imaging for Sequential Multiplexing (PRISM) to facilitate multiplexed imaging with facile, rapid exchange of imaging probes to analyze iPSC-derived cortical and motor neuron cultures that are relevant to psychiatric and neurodegenerative disease models, using over ten protein targets. Our approach permits analysis of cell differentiation, cell composition, and functional marker expression in complex stem-cell derived neural cultures. Furthermore, our approach is amenable to automation, offering in principle the ability to scale-up to dozens of protein targets and samples.

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Stöberl, Nina, Emily Maguire, Elisa Salis, Bethany Shaw et Hazel Hall-Roberts. « Human iPSC-derived glia models for the study of neuroinflammation ». Journal of Neuroinflammation 20, n^o 1 (10 octobre 2023). http://dx.doi.org/10.1186/s12974-023-02919-2.

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AbstractNeuroinflammation is a complex biological process that plays a significant role in various brain disorders. Microglia and astrocytes are the key cell types involved in inflammatory responses in the central nervous system. Neuroinflammation results in increased levels of secreted inflammatory factors, such as cytokines, chemokines, and reactive oxygen species. To model neuroinflammation in vitro, various human induced pluripotent stem cell (iPSC)-based models have been utilized, including monocultures, transfer of conditioned media between cell types, co-culturing multiple cell types, neural organoids, and xenotransplantation of cells into the mouse brain. To induce neuroinflammatory responses in vitro, several stimuli have been established that can induce responses in either microglia, astrocytes, or both. Here, we describe and critically evaluate the different types of iPSC models that can be used to study neuroinflammation and highlight how neuroinflammation has been induced and measured in these cultures.

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Khan, Mushfiquddin, Tajinder S. Dhammu, Mauhamad Baarine, Avtar K. Singh et Inderjit Singh. « Abstract WP113 : Induced Pluripotent Stem Cells Derived Neurons Ideally Serve as a Human Stroke Model of Neuronal Damage and Neuroprotective Intervention ». Stroke 48, suppl_1 (février 2017). http://dx.doi.org/10.1161/str.48.suppl_1.wp113.

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Background: Basic science research on stroke is lagging. Results from both animal and animal-derived neural cell models are, more often than not, less valid for human stroke. To address this lack of models for human stroke mechanisms, we developed induced pluripotent stem cell (iPSC)-derived human neurons and subjected them to excitotoxicity and oxygen-glucose deprivation (OGD) conditions. These conditions produced a disturbed nitric oxide (NO) metabolome similar to human stroke. A therapeutic intervention, the NO-based neuroprotective agent S-nitrosoglutathione (GSNO), was investigated for neuroprotection against excitotoxicity as well as OGD-induced neuronal nitric oxide synthase (nNOS) activation and the deleterious nNOS/peroxynitrite/calpain neurodegenerative system. Methods: The iPSC were generated using skin fibroblasts from normal adult human subjects. Neural precursor cells (NPC) were prepared using embryonic body and NPC were differentiated into neurons. Results: The differentiated iPSC neurons were positive for NeuN and β-tubulin (neuronal markers), and they responded to glutamate/NMDA-induced Ca 2+ influx. The majority of iPSC neurons (~90%) were nNOS-positive, supporting them as a suitable model for NO metabolome studies. To examine this model’s nNOS/peroxynitrite/calpain system under stroke conditions, nNOS activation (pnNOS)-dependent toxicity in the iPSC-derived neurons was observed. High expression levels were found for pnNOS Ser 1412 , 3-NT, and p25 fragment in glutamate/NMDA- and OGD-treated cells. GSNO treatment reversibly blocked glutamate/NMDA- and OGD-induced pnNOS and 3-NT expression levels. Calpain activity was also blocked. Conclusions: This study of iPSC-derived human neurons is the first of its kind to investigate excitotoxicity or OGD-mediated neurodegeneration mechanisms and therapeutic interventions under stroke conditions. The data provide evidence that the deleterious nNOS/peroxynitrite/calpain system in iPSC neurons is similar to human stroke and that GSNO treatment down regulates this deleterious system. These observations support that iPSC human neurons are a suitable model for studies on stroke-associated neurodegenerative mechanisms as well as therapeutic interventions.

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Bertucci, Taylor, Kathryn Bowles, Steven Lotz, Le Qi, Katherine Stevens, Susan K. Goderie, Susan Borden et al. « Human iPSC derived organoid models to study tau pathology ». Alzheimer's & ; Dementia 20, S6 (décembre 2024). https://doi.org/10.1002/alz.087353.

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AbstractBackgroundHuman pluripotent stem cell (hPSC)‐derived brain organoids patterned towards the cerebral cortex are valuable models of interactions occurring in vivo in cortical tissue. We and others have used these cortical organoids to model dominantly inherited FTD‐tau. While these studies have provided essential insights, cortical organoid models have yet to reach their full potential. Studies are hindered by well‐recognized hurdles: low production efficiency and high variability between individual organoids, across lines, and experiments. A protocol that consistently generates well‐patterned cerebral cortical organoids is needed.MethodsTwo key stages of the protocol were optimized: the initial growth of hPSCs and the first 6 days of differentiation. Analysis of hPSC pluripotency and organoid quality control (QC) measures at 20 day and 2 months was assessed by bulk RNAseq, qPCR and IHC for ± QC and single‐cell RNA sequencing (scRNAseq). The optimized protocol was tested across 63 lines to examine robustness.ResultsWe established a 96 Slit‐well plate method for efficient (approaching 100%), scalable, reproducible cortical organoid production. When hPSCs were cultured with controlled‐release FGF2 (FGF2Discs) and an SB431542 concentration appropriate for their TGFBR1/ALK5 expression level, organoid cortical patterning and reproducibility were significantly improved. Well‐patterned organoids included 16 neural subtypes identified by scRNA‐seq, abundant rosettes, and robust BCL11B+/TBR1+ cortical neurons at 2 months. In contrast, poorly patterned organoids contained mesendoderm‐related cells, identifiable by negative QC marker COL1A2 and/or few cortical neurons. Using this improved protocol, we demonstrate increased sensitivity to study the impact of MAPT mutations associated with FTD, and across different MAPT‐mutants, neurons exhibited common early changes in key metabolic pathways, including upregulation of oxidative phosphorylation and dampening of ceramide synthesis and sphingolipid signaling.ConclusionsVariability between different hPSC lines in the expression of key ligands and receptors contributes to the variability of success of cortical organoid production. Hence, small adjustments in the culture of hPSCs and the concentration of patterning molecules during the first 6 days of differentiation are essential to enable efficient and reproducible organoid production. This allows rigorous comparisons of disease phenotypes using many different lines from different donors and across heritable and sporadic diseases affecting the cerebral cortex.

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Rylaarsdam, Lauren, Jennifer Rakotomamonjy, Eleanor Pope et Alicia Guemez-Gamboa. « iPSC-derived models of PACS1 syndrome reveal transcriptional and functional deficits in neuron activity ». Nature Communications 15, n^o 1 (27 janvier 2024). http://dx.doi.org/10.1038/s41467-024-44989-7.

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AbstractPACS1 syndrome is a neurodevelopmental disorder characterized by intellectual disability and distinct craniofacial abnormalities resulting from a de novo p.R203W variant in phosphofurin acidic cluster sorting protein 1 (PACS1). PACS1 is known to have functions in the endosomal pathway and nucleus, but how the p.R203W variant affects developing neurons is not fully understood. Here we differentiated stem cells towards neuronal models including cortical organoids to investigate the impact of the PACS1 syndrome-causing variant on neurodevelopment. While few deleterious effects were detected in PACS1(+/R203W) neural precursors, mature PACS1(+/R203W) glutamatergic neurons exhibited impaired expression of genes involved in synaptic signaling processes. Subsequent characterization of neural activity using calcium imaging and multielectrode arrays revealed the p.R203W PACS1 variant leads to a prolonged neuronal network burst duration mediated by an increased interspike interval. These findings demonstrate the impact of the PACS1 p.R203W variant on developing human neural tissue and uncover putative electrophysiological underpinnings of disease.

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Sheu, Chia-Lin, Ullas Mony, Sihan Dai, Linhui Qiu et Vishnu Priya Veeraraghavan. « Advances in iPSC Technology in Neural Disease Modeling, Drug Screening, and Therapy ». Current Stem Cell Research & ; Therapy 18 (8 juin 2023). http://dx.doi.org/10.2174/1574888x18666230608105703.

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Abstract: Neurodegenerative disorders (NDs) including Alzheimer’s Disease, Parkinson’s Disease, Amyotrophic Lateral Sclerosis (ALS), and Huntington's disease are all incurable and can only be managed with drugs for the associated symptoms. Animal models of human illnesses help to advance our understanding of the pathogenic processes of diseases. Understanding the pathogenesis as well as drug screening using appropriate disease models of neurodegenerative diseases (NDs) are vital for identifying novel therapies. Human-derived induced pluripotent stem cell (iPSC) models can be an efficient model to create disease in a dish and thereby can proceed with drug screening and identifying appropriate drugs. This technology has many benefits, including efficient reprogramming and regeneration potential, multidirectional differentiation, and the lack of ethical concerns, which open up new avenues for studying neurological illnesses in greater depth. The review mainly focuses on the use of iPSC technology in neuronal disease modeling, drug screening, and cell therapy.

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Spathopoulou, Angeliki, Frank Edenhofer et Lisa Fellner. « Targeting α-Synuclein in Parkinson's Disease by Induced Pluripotent Stem Cell Models ». Frontiers in Neurology 12 (25 janvier 2022). http://dx.doi.org/10.3389/fneur.2021.786835.

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Parkinson's disease (PD) is a progressive, neurodegenerative disorder characterized by motor and non-motor symptoms. To date, no specific treatment to halt disease progression is available, only medication to alleviate symptoms can be prescribed. The main pathological hallmark of PD is the development of neuronal inclusions, positive for α-synuclein (α-syn), which are termed Lewy bodies (LBs) or Lewy neurites. However, the cause of the inclusion formation and the loss of neurons remain largely elusive. Various genetic determinants were reported to be involved in PD etiology, including SNCA, DJ-1, PRKN, PINK1, LRRK2, and GBA. Comprehensive insights into pathophysiology of PD critically depend on appropriate models. However, conventional model organisms fall short to faithfully recapitulate some features of this complex disease and as a matter-of-fact access to physiological tissue is limiting. The development of disease models replicating PD that are close to human physiology and dynamic enough to analyze the underlying molecular mechanisms of disease initiation and progression, as well as the generation of new treatment options, is an important and overdue step. Recently, the establishment of induced pluripotent stem cell (iPSC)-derived neural models, particularly from genetic PD-variants, developed into a promising strategy to investigate the molecular mechanisms regarding formation of inclusions and neurodegeneration. As these iPSC-derived neurons can be generated from accessible biopsied samples of PD patients, they carry pathological alterations and enable the possibility to analyze the differences compared to healthy neurons. This review focuses on iPSC models carrying genetic PD-variants of α-syn that will be especially helpful in elucidating the pathophysiological mechanisms of PD. Furthermore, we discuss how iPSC models can be instrumental in identifying cellular targets, potentially leading to the development of new therapeutic treatments. We will outline the enormous potential, but also discuss the limitations of iPSC-based α-syn models.

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Pedroza, Albert J., Samantha Churovich, Nobu Yokoyama, Ken Nakamura, Cristiana Iosef Husted et Michael P. Fischbein. « Abstract 14908 : Anatomic Validation of Induced Pluripotent Stem Cell-derived Aortic Smooth Muscle Cell Model of Loeys Dietz Syndrome ». Circulation 142, Suppl_3 (17 novembre 2020). http://dx.doi.org/10.1161/circ.142.suppl_3.14908.

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Introduction: Mutations in TGF-beta (TGF-ß) signaling genes lead to aortic root aneurysm in Loeys Dietz syndrome (LDS). Smooth muscle cells (SMCs) in the proximal aorta develop from two embryologic origins: second heart field (SHF) and neural crest (NC). Induced pluripotent stem cell (iPSC) models simulate these lineages, but direct correlation to clinical disease is lacking. Hypothesis: iPSC-derived SMCs accurately model lineage-specific aortopathy in LDS. Methods: We generated SMC lines from root and ascending aortic surgical tissue and iPSC-derived SMCs through SHF and NC-specific pathways from an LDS patient ( TGFBR1 mutation). Lineage-specific TGF-ß responses were determined by western blot/ELISA. RNA sequencing and RT-PCR identified SMC transcriptomes. Results: Aortic root SMCs showed greater canonical TGF-ß activation (p-SMAD2/3) versus ascending at baseline and with TGF-ß stimulation ( Figure ). Synonymous results were seen in SHF versus NC SMCs from the iPSC pathway. RNAseq identified 1,600 differentially expressed genes between iPSC lineages, including altered TGF-ß receptor and ligand expression profiles. Primary aortic lines validated iPSC data: root SMCs showed enriched TGF-ß receptor 1/2/3 expression (1.7-, 3.9- and 5.9-fold) while ascending SMCs overexpressed TGFB1 and TGFB2 ligands (1.8- and 3.5-fold). Despite discordant TGF-ß activation, SMC contractile gene expression was similar between lineages in aortic and iPSC-SMCs, suggesting alternative downstream effects in LDS aneurysm. Conclusion: iPSC-derived SMCs effectively model lineage-specific aortic root aneurysm pathology, validating this model as a tool for mechanistic testing and therapy discovery.

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Gorgogietas, Vyron, Bahareh Rajaei, Chae Heeyoung, Bruno J. Santacreu, Sandra Marín-Cañas, Paraskevi Salpea, Toshiaki Sawatani et al. « GLP-1R agonists demonstrate potential to treat Wolfram syndrome in human preclinical models ». Diabetologia, 30 mars 2023. http://dx.doi.org/10.1007/s00125-023-05905-8.

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Abstract Aims/hypothesis Wolfram syndrome is a rare autosomal recessive disorder caused by pathogenic variants in the WFS1 gene. It is characterised by insulin-dependent diabetes mellitus, optic nerve atrophy, diabetes insipidus, hearing loss and neurodegeneration. Considering the unmet treatment need for this orphan disease, this study aimed to evaluate the therapeutic potential of glucagon-like peptide 1 receptor (GLP-1R) agonists under wolframin (WFS1) deficiency with a particular focus on human beta cells and neurons. Methods The effect of the GLP-1R agonists dulaglutide and exenatide was examined in Wfs1 knockout mice and in an array of human preclinical models of Wolfram syndrome, including WFS1-deficient human beta cells, human induced pluripotent stem cell (iPSC)-derived beta-like cells and neurons from control individuals and individuals affected by Wolfram syndrome, and humanised mice. Results Our study shows that the long-lasting GLP-1R agonist dulaglutide reverses impaired glucose tolerance in WFS1-deficient mice, and that exenatide and dulaglutide improve beta cell function and prevent apoptosis in different human WFS1-deficient models including iPSC-derived beta cells from people with Wolfram syndrome. Exenatide improved mitochondrial function, reduced oxidative stress and prevented apoptosis in Wolfram syndrome iPSC-derived neural precursors and cerebellar neurons. Conclusions/interpretation Our study provides novel evidence for the beneficial effect of GLP-1R agonists on WFS1-deficient human pancreatic beta cells and neurons, suggesting that these drugs may be considered as a treatment for individuals with Wolfram syndrome. Graphical abstract

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Roth, Julien G., Kristin L. Muench, Aditya Asokan, Victoria M. Mallett, Hui Gai, Yogendra Verma, Stephen Weber et al. « 16p11.2 microdeletion imparts transcriptional alterations in human iPSC-derived models of early neural development ». eLife 9 (10 novembre 2020). http://dx.doi.org/10.7554/elife.58178.

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Microdeletions and microduplications of the 16p11.2 chromosomal locus are associated with syndromic neurodevelopmental disorders and reciprocal physiological conditions such as macro/microcephaly and high/low body mass index. To facilitate cellular and molecular investigations into these phenotypes, 65 clones of human induced pluripotent stem cells (hiPSCs) were generated from 13 individuals with 16p11.2 copy number variations (CNVs). To ensure these cell lines were suitable for downstream mechanistic investigations, a customizable bioinformatic strategy for the detection of random integration and expression of reprogramming vectors was developed and leveraged towards identifying a subset of ‘footprint’-free hiPSC clones. Transcriptomic profiling of cortical neural progenitor cells derived from these hiPSCs identified alterations in gene expression patterns which precede morphological abnormalities reported at later neurodevelopmental stages. Interpreting clinical information—available with the cell lines by request from the Simons Foundation Autism Research Initiative—with this transcriptional data revealed disruptions in gene programs related to both nervous system function and cellular metabolism. As demonstrated by these analyses, this publicly available resource has the potential to serve as a powerful medium for probing the etiology of developmental disorders associated with 16p11.2 CNVs.

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Cheng, Chialin, Surya A. Reis, Emily T. Adams, Daniel M. Fass, Steven P. Angus, Timothy J. Stuhlmiller, Jared Richardson et al. « High-content image-based analysis and proteomic profiling identifies Tau phosphorylation inhibitors in a human iPSC-derived glutamatergic neuronal model of tauopathy ». Scientific Reports 11, n^o 1 (23 août 2021). http://dx.doi.org/10.1038/s41598-021-96227-5.

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AbstractMutations in MAPT (microtubule-associated protein tau) cause frontotemporal dementia (FTD). MAPT mutations are associated with abnormal tau phosphorylation levels and accumulation of misfolded tau protein that can propagate between neurons ultimately leading to cell death (tauopathy). Recently, a p.A152T tau variant was identified as a risk factor for FTD, Alzheimer's disease, and synucleinopathies. Here we used induced pluripotent stem cells (iPSC) from a patient carrying this p.A152T variant to create a robust, functional cellular assay system for probing pathophysiological tau accumulation and phosphorylation. Using stably transduced iPSC-derived neural progenitor cells engineered to enable inducible expression of the pro-neural transcription factor Neurogenin 2 (Ngn2), we generated disease-relevant, cortical-like glutamatergic neurons in a scalable, high-throughput screening compatible format. Utilizing automated confocal microscopy, and an advanced image-processing pipeline optimized for analysis of morphologically complex human neuronal cultures, we report quantitative, subcellular localization-specific effects of multiple kinase inhibitors on tau, including ones under clinical investigation not previously reported to affect tau phosphorylation. These results demonstrate the potential for using patient iPSC-derived ex vivo models of tauopathy as genetically accurate, disease-relevant systems to probe tau biochemistry and support the discovery of novel therapeutics for tauopathies.

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