10 results on '"Neyrinck K"'
Search Results
2. In vitro modeling of dysfunctional glial cells in neurodegenerative diseases using human pluripotent stem cells
- Author
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García León, Juan Antonio, Eggermont, K, Neyrinck, K, Davila, Jose Carlos, Vitorica, Javier, Verfaillie, C, and Gutierrez-Perez, Antonia
- Abstract
Most neurodegenerative diseases are characterized by a complex and mostly still unresolved pathology. This fact, together with the lack of reliable models, have precluded the development of effective therapies counteracting the disease progression. In the past few years, several studies have evidenced that lack of proper functionality of glial cells (astrocytes, microglia and oligodendrocytes) has a key role in the pathology of several neurodegenerative conditions including Alzheimer´s disease, amyotrophic lateral sclerosis and multiple sclerosis among others. However, this glial dysfunction is poorly modelled by available animal models, and we hypothesize that patientderived cells can serve as a better platform where to study this glial dysfunction. In this sense, human pluripotent stem cells (hPSCs) has revolutionized the field allowing the generation of disease-relevant neural cell types that can be used for disease modelling, drug screening and, possibly, cell transplantation purposes. In the case of the generation of oligodendrocytes (OLs) from hPSCs, we have developed a fast and robust protocol to generate surface antigen O4-positive (O4+) and myelin basic protein-positive OLs from hPSCs in only 22 days, including from patients with multiple sclerosis or amyotrophic lateral sclerosis. The generated cells resemble primary human OLs at the transcriptome level and can myelinate neurons in vivo. Using in vitro OLneuron co-cultures, effective myelination of neurons can also be demonstrated. This platform is being translated as well to the generation of the other glial cell types, allowing the derivation of patient-specific glial cells where to model disease-specific dysfunction. This methodology can be used for elucidating pathogenic pathways associated with neurodegeneration and to identify therapeutic targets susceptible of drug modulation, contributing to the development of novel and effective drugs for these devastating disorders. Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. Supported by PI18/01557 (to AG) and P18/1556 (to JV) grants from ISCiii of Spain co-financed by FEDER funds from European Union, and PI-0276-2018 grant (to JAGL) from Consejeria de Salud of Junta de Andalucia. JAGL held a postdoctoral contract from the I Research Plan Propio of the University of Malaga. CV and KE were supported by IWT-SBO-150031-iPSCAF and the Thierry Lathran Foundation grant – ALS-OL, and KN by FWO1166518N
- Published
- 2019
3. [Psychiatric disorders in velo-cardio-facial syndrome]
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Maesen A, Stephan Claes, and Neyrinck K
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Adult ,Male ,Alcoholism ,Phenotype ,Chromosomes, Human, Pair 22 ,Face ,Mental Disorders ,DiGeorge Syndrome ,Humans ,Abnormalities, Multiple ,Sequence Deletion - Abstract
Velo-cardio-facial syndrome (vcfs) is a congenital disorder with a markedly variable clinical expression. The majority of those affected have cognitive-behavioural symptoms and psychiatric problems. Most of the somatic characteristics can be treated effectively. The quality of life of patients with vcfs is therefore determined largely by cognitive and behavioural symptoms, including the increased risk of psychiatric disorders. On the basis of a case-study featuring a 41-year-old vcfs patient and by reviewing the literature we describe the psychiatric disorders that can occur in conjunction with this syndrome.
- Published
- 2010
4. Local Analytic Models For Families Of Hyperbolic Vector Fields
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BONCKAERT, Patrick and Neyrinck, K.
- Abstract
We look for analytic models near hyperbolic singularities of families of real analytic vector fields Xe. The interesting case deals with saddles, since for sources or sinks we have the results of Poincar´e [1, 2]. For families we cannot use the Siegel theorem since the condition on the small divisors is fragile. Even on the formal level (i.e. power series) the number of resonances between the eigenvalues is infinite for a family: for instance in the case of a planar saddle this comes to the density of the rationals in R. One option is to use a Ck (k < 1) normal form for the family [5]. Here we want to remain within the analytic category, and have to allow a less simplified form. A first standard simplification is to use stable and unstable manifolds, and to ’straighten’ them, i.e. to write the vector field such that these are linear subspaces. The fact that these invariant manifolds are analytic and depend analytically on the parameter will also follow from the results in this paper. The normal form we aim at will be moreover be ’as flat as desired’ along these invariant manifolds if there are no low order resonances for X0. This approach can already be found in [3, 11, 12] and we extend the results in [12], on which our methods are inspired. Even though in this paper we confine ourselves to the case of a family of vector fields, we can prove similar results for a family of diffeomorphisms [7]. We shall also prove that possible symmetries are preserved in our local analytic model and by the changes of variables.
- Published
- 2006
5. Correction to: Disruption of MAM integrity in mutant FUS oligodendroglial progenitors from hiPSCs.
- Author
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Zhu Y, Burg T, Neyrinck K, Vervliet T, Nami F, Vervoort E, Ahuja K, Sassano ML, Chai YC, Tharkeshwar AK, De Smedt J, Hu H, Bultynck G, Agostinis P, Swinnen JV, Van Den Bosch L, da Costa RFM, and Verfaillie C
- Published
- 2024
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6. Disruption of MAM integrity in mutant FUS oligodendroglial progenitors from hiPSCs.
- Author
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Zhu Y, Burg T, Neyrinck K, Vervliet T, Nami F, Vervoort E, Ahuja K, Sassano ML, Chai YC, Tharkeshwar AK, De Smedt J, Hu H, Bultynck G, Agostinis P, Swinnen JV, Van Den Bosch L, da Costa RFM, and Verfaillie C
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- Humans, Motor Neurons metabolism, Mutation, Oligodendroglia metabolism, RNA-Binding Protein FUS genetics, RNA-Binding Protein FUS metabolism, Amyotrophic Lateral Sclerosis pathology, Induced Pluripotent Stem Cells metabolism
- Abstract
Amyotrophic lateral sclerosis (ALS) is a rapidly progressive and fatal neurodegenerative disorder, characterized by selective loss of motor neurons (MNs). A number of causative genetic mutations underlie the disease, including mutations in the fused in sarcoma (FUS) gene, which can lead to both juvenile and late-onset ALS. Although ALS results from MN death, there is evidence that dysfunctional glial cells, including oligodendroglia, contribute to neurodegeneration. Here, we used human induced pluripotent stem cells (hiPSCs) with a R521H or a P525L mutation in FUS and their isogenic controls to generate oligodendrocyte progenitor cells (OPCs) by inducing SOX10 expression from a TET-On SOX10 cassette. Mutant and control iPSCs differentiated efficiently into OPCs. RNA sequencing identified a myelin sheath-related phenotype in mutant OPCs. Lipidomic studies demonstrated defects in myelin-related lipids, with a reduction of glycerophospholipids in mutant OPCs. Interestingly, FUS
R521H OPCs displayed a decrease in the phosphatidylcholine/phosphatidylethanolamine ratio, known to be associated with maintaining membrane integrity. A proximity ligation assay further indicated that mitochondria-associated endoplasmic reticulum membranes (MAM) were diminished in both mutant FUS OPCs. Moreover, both mutant FUS OPCs displayed increased susceptibility to ER stress when exposed to thapsigargin, and exhibited impaired mitochondrial respiration and reduced Ca2+ signaling from ER Ca2+ stores. Taken together, these results demonstrate a pathological role of mutant FUS in OPCs, causing defects in lipid metabolism associated with MAM disruption manifested by impaired mitochondrial metabolism with increased susceptibility to ER stress and with suppressed physiological Ca2+ signaling. As such, further exploration of the role of oligodendrocyte dysfunction in the demise of MNs is crucial and will provide new insights into the complex cellular mechanisms underlying ALS., (© 2024. The Author(s).)- Published
- 2024
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7. SOX9-induced Generation of Functional Astrocytes Supporting Neuronal Maturation in an All-human System.
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Neyrinck K, Van Den Daele J, Vervliet T, De Smedt J, Wierda K, Nijs M, Vanbokhoven T, D'hondt A, Planque M, Fendt SM, Shih PY, Seibt F, Almenar JP, Kreir M, Kumar D, Broccoli V, Bultynck G, Ebneth A, Cabrera-Socorro A, and Verfaillie C
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- Animals, Humans, Induced Pluripotent Stem Cells cytology, Mice, Neurogenesis physiology, Neurons cytology, SOX9 Transcription Factor metabolism, Astrocytes cytology, Neural Stem Cells physiology
- Abstract
Astrocytes, the main supportive cell type of the brain, show functional impairments upon ageing and in a broad spectrum of neurological disorders. Limited access to human astroglia for pre-clinical studies has been a major bottleneck delaying our understanding of their role in brain health and disease. We demonstrate here that functionally mature human astrocytes can be generated by SOX9 overexpression for 6 days in pluripotent stem cell (PSC)-derived neural progenitor cells. Inducible (i)SOX9-astrocytes display functional properties comparable to primary human astrocytes comprising glutamate uptake, induced calcium responses and cytokine/growth factor secretion. Importantly, electrophysiological properties of iNGN2-neurons co-cultured with iSOX9-astrocytes are indistinguishable from gold-standard murine primary cultures. The high yield, fast timing and the possibility to cryopreserve iSOX9-astrocytes without losing functional properties makes them suitable for scaled-up production for high-throughput analyses. Our findings represent a step forward to an all-human iPSC-derived neural model for drug development in neuroscience and towards the reduction of animal use in biomedical research., (© 2021. The Author(s).)
- Published
- 2021
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8. Single Transcription Factor-Based Differentiation Allowing Fast and Efficient Oligodendrocyte Generation via SOX10 Overexpression.
- Author
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Neyrinck K and García-León JA
- Subjects
- Cell Culture Techniques, Cells, Cultured, Cloning, Molecular, Gene Order, Genetic Vectors administration & dosage, Genetic Vectors biosynthesis, Genetic Vectors genetics, Humans, Immunomagnetic Separation, Lentivirus genetics, Neurogenesis, Cell Differentiation genetics, Gene Expression, Oligodendroglia cytology, Oligodendroglia metabolism, SOXE Transcription Factors genetics, Transcription Factors genetics
- Abstract
Oligodendrocytes are the main glial cell type in the central nervous system supporting the axonal part of neurons via myelin and lactate delivery. Both the conductive myelin formation and the energy support via lactate can be affected in diseases, such as multiple sclerosis and amyotrophic lateral sclerosis, respectively. Therefore, human disease modeling is needed to gain more mechanistic insights to drive drug discovery research. Here, patient-derived induced pluripotent stem cells (iPSCs) serve as a necessary tool providing an infinite cell source for patient-specific disease modeling, which allows investigation of oligodendrocyte involvement in human disease.Small molecule-based differentiation protocols to generate oligodendrocytes from pluripotent stem cells can last more than 90 days. Here, we provide a transcription factor-based, fast and efficient protocol for generating O4
+ oligodendrocytes in just 20-24 days. After a neural induction phase of 8-12 days, SOX10 is overexpressed either with the use of lentiviral vectors or via engineered iPSCs, which inducibly overexpress SOX10 after doxycycline addition. Using this last method, a pure O4+ cell population is achieved after keeping the SOX10-overexpressing neural stem cells in culture for an additional 10 days. Furthermore, these O4+ cells can be co-cultured with iPSC-derived cortical neurons in 384-well format, allowing pro-myelinating drug screens. In conclusion, we provide a fast and efficient oligodendrocyte differentiation protocol allowing both in vitro human disease modeling and a high-throughput co-culture system for drug discovery., (© 2021. Springer Science+Business Media, LLC, part of Springer Nature.)- Published
- 2021
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9. Generation of oligodendrocytes and establishment of an all-human myelinating platform from human pluripotent stem cells.
- Author
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García-León JA, García-Díaz B, Eggermont K, Cáceres-Palomo L, Neyrinck K, Madeiro da Costa R, Dávila JC, Baron-Van Evercooren A, Gutiérrez A, and Verfaillie CM
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- Cell Culture Techniques methods, Cell Line, Humans, Myelin Basic Protein analysis, Myelin Basic Protein metabolism, Neural Stem Cells metabolism, Oligodendroglia metabolism, Pluripotent Stem Cells metabolism, Myelin Sheath metabolism, Neural Stem Cells cytology, Neurogenesis, Oligodendroglia cytology, Pluripotent Stem Cells cytology
- Abstract
Oligodendrocytes (OLs) are responsible for myelin production and metabolic support of neurons. Defects in OLs are crucial in several neurodegenerative diseases including multiple sclerosis (MS) and amyotrophic lateral sclerosis (ALS). This protocol describes a method to generate oligodendrocyte precursor cells (OPCs) from human pluripotent stem cells (hPSCs) in only ~20 d, which can subsequently myelinate neurons, both in vitro and in vivo. To date, OPCs have been derived from eight different hPSC lines including those derived from patients with spontaneous and familial forms of MS and ALS, respectively. hPSCs, fated for 8 d toward neural progenitors, are transduced with an inducible lentiviral vector encoding for SOX10. The addition of doxycycline for 10 d results in >60% of cells being O4-expressing OPCs, of which 20% co-express the mature OL marker myelin basic protein (MBP). The protocol also describes an alternative for viral transduction, by incorporating an inducible SOX10 in the safe harbor locus AAVS1, yielding ~100% pure OPCs. O4
+ OPCs can be purified and either cryopreserved or used for functional studies. As an example of the type of functional study for which the derived cells could be used, O4+ cells can be co-cultured with maturing hPSC-derived neurons in 96/384-well-format plates, allowing the screening of pro-myelinating compounds.- Published
- 2020
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10. The human somatostatin receptor type 2 as an imaging and suicide reporter gene for pluripotent stem cell-derived therapy of myocardial infarction.
- Author
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Neyrinck K, Breuls N, Holvoet B, Oosterlinck W, Wolfs E, Vanbilloen H, Gheysens O, Duelen R, Gsell W, Lambrichts I, Himmelreich U, Verfaillie CM, Sampaolesi M, and Deroose CM
- Subjects
- Animals, Cell Line, Female, Genes, Reporter, Human Embryonic Stem Cells cytology, Human Embryonic Stem Cells metabolism, Humans, Luciferases genetics, Luciferases metabolism, Mice, Mice, Nude, Myocardial Infarction diagnostic imaging, Myocytes, Cardiac cytology, Myocytes, Cardiac metabolism, Octreotide analogs & derivatives, Organometallic Compounds, Positron-Emission Tomography, Radiopharmaceuticals, Receptors, Somatostatin genetics, Stem Cell Transplantation adverse effects, Teratoma etiology, Human Embryonic Stem Cells transplantation, Myocardial Infarction therapy, Receptors, Somatostatin metabolism, Stem Cell Transplantation methods, Teratoma diagnostic imaging
- Abstract
Rationale: Pluripotent stem cells (PSCs) are being investigated as a cell source for regenerative medicine since they provide an infinitive pool of cells that are able to differentiate towards every cell type of the body. One possible therapeutic application involves the use of these cells to treat myocardial infarction (MI), a condition where billions of cardiomyocytes (CMs) are lost. Although several protocols have been developed to differentiate PSCs towards CMs, none of these provide a completely pure population, thereby still posing a risk for neoplastic teratoma formation. Therefore, we developed a strategy to (i) monitor cell behavior noninvasively via site-specific integration of firefly luciferase (Fluc) and the human positron emission tomography (PET) imaging reporter genes, sodium iodide symporter (hNIS) and somatostatin receptor type 2 (hSSTr2), and (ii) perform hSSTr2-mediated suicide gene therapy via the clinically used radiopharmacon
177 Lu-DOTATATE. Methods: Human embryonic stem cells (ESCs) were gene-edited via zinc finger nucleases to express Fluc and either hNIS or hSSTr2 in the safe harbor locus, adeno-associated virus integration site 1. Firstly, these cells were exposed to 4.8 MBq177 Lu-DOTATATE in vitro and cell survival was monitored via bioluminescence imaging (BLI). Afterwards, hNIS+ and hSSTr2+ ESCs were transplanted subcutaneously and teratomas were allowed to form. At day 59, baseline124 I and68 Ga-DOTATATE PET and BLI scans were performed. The day after, animals received either saline or 55 MBq177 Lu-DOTATATE. Weekly BLI scans were performed, accompanied by124 I and68 Ga-DOTATATE PET scans at days 87 and 88, respectively. Finally, hSSTr2+ ESCs were differentiated towards CMs and transplanted intramyocardially in the border zone of an infarct that was induced by left anterior descending coronary artery ligation. After transplantation, the animals were monitored via BLI and PET, while global cardiac function was evaluated using cardiac magnetic resonance imaging. Results: Teratoma growth of both hNIS+ and hSSTr2+ ESCs could be followed noninvasively over time by both PET and BLI. After177 Lu-DOTATATE administration, successful cell killing of the hSSTr2+ ESCs was achieved both in vitro and in vivo , indicated by reductions in total tracer lesion uptake, BLI signal and teratoma volume. As undifferentiated hSSTr2+ ESCs are not therapeutically relevant, they were differentiated towards CMs and injected in immune-deficient mice with a MI. Long-term cell survival could be monitored without uncontrolled cell proliferation. However, no improvement in the left ventricular ejection fraction was observed. Conclusion: We developed isogenic hSSTr2-expressing ESCs that allow noninvasive cell monitoring in the context of PSC-derived regenerative therapy. Furthermore, we are the first to use the hSSTr2 not only as an imaging reporter gene, but also as a suicide mechanism for radionuclide therapy in the setting of PSC-derived cell treatment., Competing Interests: Competing interests: Katrien Neyrinck works as an aspirant for the FWO, while both Natacha Breuls and Bryan Holvoet work under an SB-grant of the FWO. Christophe M. Deroose is a consultant for Advanced Accelerator Applications (AAA) and Novartis, who hold rights to commercialize 177Lu-DOTATATE. He is also a consultant for Sirtex, Bayer, Ipsen and Terumo. He received travel grants from GE Healthcare. No other potential conflict of interest relevant to this article was reported.- Published
- 2018
- Full Text
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