175 results on '"Ebneth A"'
Search Results
2. Pharmacological characterization of mutant huntingtin aggregate-directed PET imaging tracer candidates
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Frank Herrmann, Manuela Hessmann, Sabine Schaertl, Karola Berg-Rosseburg, Christopher J Brown, Galina Bursow, Anass Chiki, Andreas Ebneth, Miriam Gehrmann, Nicole Hoeschen, Madlen Hotze, Stefanie Jahn, Peter D Johnson, Vinod Khetarpal, Alex Kiselyov, Karsten Kottig, Stefanie Ladewig, Hilal Lashuel, Sven Letschert, Matthew R Mills, Kathrin Petersen, Michael E Prime, Christoph Scheich, Gerhard Schmiedel, John Wityak, Longbin Liu, Celia Dominguez, Ignacio Muñoz-Sanjuán, and Jonathan A Bard
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Medicine ,Science - Abstract
Abstract Huntington’s disease (HD) is caused by a CAG trinucleotide repeat expansion in the first exon of the huntingtin (HTT) gene coding for the huntingtin (HTT) protein. The misfolding and consequential aggregation of CAG-expanded mutant HTT (mHTT) underpin HD pathology. Our interest in the life cycle of HTT led us to consider the development of high-affinity small-molecule binders of HTT oligomerized/amyloid-containing species that could serve as either cellular and in vivo imaging tools or potential therapeutic agents. We recently reported the development of PET tracers CHDI-180 and CHDI-626 as suitable for imaging mHTT aggregates, and here we present an in-depth pharmacological investigation of their binding characteristics. We have implemented an array of in vitro and ex vivo radiometric binding assays using recombinant HTT, brain homogenate-derived HTT aggregates, and brain sections from mouse HD models and humans post-mortem to investigate binding affinities and selectivity against other pathological proteins from indications such as Alzheimer’s disease and spinocerebellar ataxia 1. Radioligand binding assays and autoradiography studies using brain homogenates and tissue sections from HD mouse models showed that CHDI-180 and CHDI-626 specifically bind mHTT aggregates that accumulate with age and disease progression. Finally, we characterized CHDI-180 and CHDI-626 regarding their off-target selectivity and binding affinity to beta amyloid plaques in brain sections and homogenates from Alzheimer’s disease patients.
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- 2021
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3. The EBiSC iPSC bank for disease studies
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Steeg, Rachel, Neubauer, Julia C., Müller, Sabine C., Ebneth, Andreas, and Zimmermann, Heiko
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- 2020
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4. Deploying human pluripotent stem cells to treat central nervous system disorders: facts, challenges and realising the potential
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Allsopp, Timothy E, Ebneth, Andreas, and Cabrera-Socorro, Alfredo
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- 2019
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5. Small Molecule Binding to Alzheimer Risk Factor CD33 Promotes Aβ Phagocytosis
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Miles, Luke A., Hermans, Stefan J., Crespi, Gabriela A.N., Gooi, Jonathan H., Doughty, Larissa, Nero, Tracy L., Markulić, Jasmina, Ebneth, Andreas, Wroblowski, Berthold, Oehlrich, Daniel, Trabanco, Andrés A., Rives, Marie-Laure, Royaux, Ines, Hancock, Nancy C., and Parker, Michael W.
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- 2019
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6. Development of a fully human assay combining NGN2-inducible neurons co-cultured with iPSC-derived astrocytes amenable for electrophysiological studies
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Pei-Yu Shih, Mohamed Kreir, Devesh Kumar, Frederik Seibt, Francisco Pestana, Benjamin Schmid, Bjørn Holst, Christian Clausen, Rachel Steeg, Benjamin Fischer, Juan Pita-Almenar, Andreas Ebneth, and Alfredo Cabrera-Socorro
- Subjects
Biology (General) ,QH301-705.5 - Abstract
Neurogenin 2 encodes a neural-specific transcription factor (NGN2) able to drive neuronal fate on somatic and stem cells. NGN2 is expressed in neural progenitors within the developing central and peripheral nervous systems. Overexpression of NGN2 in human induced pluripotent stem cells (hiPSCs) or human embryonic stem cells has been shown to efficiently trigger conversion to neurons. Here we describe two gene-edited hiPSC lines harbouring a doxycycline (DOX)-inducible cassette in the AAVS1 locus driving expression of NGN2 (BIONi010-C-13) or NGN2-T2A-GFP (BIONi010-C-15). By introducing NGN2-expressing cassette, we reduce variability associated with conventional over-expression methods such as viral transduction, making these lines amenable for scale-up production and screening processes. DOX-treated hiPSCs convert to neural phenotype within one week and display the expression of structural neuronal markers such as Beta-III tubulin and tau. We performed functional characterization of NGN2-neurons co-cultured with hiPSC-derived astrocytes in a “fully-humanized” set up. Passive properties of NGN2-neurons were indistinguishable from mouse primary cells while displaying variable activity in extracellular recordings performed in multi-electrode arrays (MEAs). We demonstrate that hiPSC-derived astrocytes and neurons can be co-cultured and display functional properties comparable to the gold standard used in electrophysiology. Both lines are globally available via EBiSC repository at https://cells.ebisc.org/.
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- 2021
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7. Small Molecule Binding to Alzheimer Risk Factor CD33 Promotes Aβ Phagocytosis
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Luke A. Miles, Stefan J. Hermans, Gabriela A.N. Crespi, Jonathan H. Gooi, Larissa Doughty, Tracy L. Nero, Jasmina Markulić, Andreas Ebneth, Berthold Wroblowski, Daniel Oehlrich, Andrés A. Trabanco, Marie-Laure Rives, Ines Royaux, Nancy C. Hancock, and Michael W. Parker
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Science - Abstract
Summary: Polymorphism in the microglial receptor CD33 gene has been linked to late-onset Alzheimer disease (AD), and reduced expression of the CD33 sialic acid-binding domain confers protection. Thus, CD33 inhibition might be an effective therapy against disease progression. Progress toward discovery of selective CD33 inhibitors has been hampered by the absence of an atomic resolution structure. We report here the crystal structures of CD33 alone and bound to a subtype-selective sialic acid mimetic called P22 and use them to identify key binding residues by site-directed mutagenesis and binding assays to reveal the molecular basis for its selectivity toward sialylated glycoproteins and glycolipids. We show that P22, when presented on microparticles, increases uptake of the toxic AD peptide, amyloid-β (Aβ), into microglial cells. Thus, the sialic acid-binding site on CD33 is a promising pharmacophore for developing therapeutics that promote clearance of the Aβ peptide that is thought to cause AD. : Molecular Structure; Neuroscience; Molecular Neuroscience; Components of the Immune System; Protein Structure Aspects Subject Areas: Molecular Structure, Neuroscience, Molecular Neuroscience, Components of the Immune System, Protein Structure Aspects
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- 2019
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8. Generation of a set of isogenic iPSC lines carrying all APOE genetic variants (Ɛ2/Ɛ3/Ɛ4) and knock-out for the study of APOE biology in health and disease
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Benjamin Schmid, Bjørn Holst, Christian Clausen, Lamiaa Bahnassawy, Peter Reinhardt, Margot H.M. Bakker, Eva Díaz-Guerra, Carlos Vicario, Pamela V. Martino-Adami, Michaela Thoenes, Alfredo Ramirez, Klaus Flissbach, Clara Grezella, Oliver Brüstle, Michael Peitz, Andreas Ebneth, and Alfredo Cabrera-Socorro
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Biology (General) ,QH301-705.5 - Abstract
APOE genotype is the strongest genetic risk factor for Alzheimer’s Disease (AD). The low degree of homology between mouse and human APOE is a concerning issue in preclinical models currently used to study the role of this gene in AD pathophysiology. A key objective of ADAPTED (Alzheimer's Disease Apolipoprotein Pathology for Treatment Elucidation and Development) project was to generate in vitro models that better recapitulate human APOE biology. We describe a new set of induced pluripotent stem cells (iPSC) lines carrying common APOE variants (Ɛ2, Ɛ3, and Ɛ3/Ɛ4) and a knock-out isogenic to the parental APOE Ɛ4/Ɛ4 line (UKBi011-A).
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- 2021
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9. Generation of two gene edited iPSC-lines carrying a DOX-inducible NGN2 expression cassette with and without GFP in the AAVS1 locus
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Benjamin Schmid, Bjørn Holst, Ulla Poulsen, Ida Jørring, Christian Clausen, M. Rasmussen, Ulrike A. Mau-Holzmann, Rachel Steeg, Hugh Nuthall, A. Ebneth, and A. Cabrera-Socorro
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Biology (General) ,QH301-705.5 - Abstract
Neurog2 is the gene encoding the neuronal transcription factor NGN2, which can convert stem cells into functional neurons in a fast and efficient way. Here we report the generation of two iPS cell lines, where DOX inducible constructs of neurog2 either without or with T2A-eGFP were inserted into the safe-site locus AAVS1. These iPS cell lines, BIONi010-C-13 and BIONi010-C-15, respectively, stay pluripotent without DOX but differentiate to (GFP positive) neurons when DOX is added without the need of differentiation factors.
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- 2021
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10. The EBiSC iPSC bank for disease studies
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Rachel Steeg, Julia C. Neubauer, Sabine C. Müller, Andreas Ebneth, and Heiko Zimmermann
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iPSC ,EBiSC ,Induced ,Pluripotent ,Stem ,Cell ,Biology (General) ,QH301-705.5 - Abstract
The European Bank for induced Pluripotent Stem Cells (EBiSC), a non-profit repository for storage, banking, Quality Control (QC) and subsequent distribution of research-grade human induced Pluripotent Stem Cell (iPSC) lines, has centralised iPSC lines generated internationally across >35 disease areas and made them available to users via the EBiSC Catalogue, for research use (cells.ebisc.org/). Comprehensive datasets are accessible prior to purchase detailing the disease background of the original tissue sample, background of iPSC reprogramming and cell line characterisation data. EBiSC also performs robust QC screening to ensure supply of reliable, well-characterised iPSC lines, compliant with ISO9001:2015 principles. Whole Genome Sequencing data for specific iPSC lines can be downloaded from the European Genome Archive, subject to application to the EBiSC Data Access Committee. The EBiSC Access and Use Agreement, required to be completed prior to shipping, can be downloaded from the website along with specific Cell Line Information Packs; together these documents clarify how EBiSC lines can be used for research and detail any specific Third Party Obligations and/or restrictions for use which may apply. A protocol for how to culture and monitor iPSC lines including implementation of routine cell line screening is also available. A second project phase will continue collecting iPSC lines generated internationally, provide iPSC derived differentiated products using improved automation strategies for upscaling and develop the current services provided by EBiSC, including iPSC reprogramming, gene-editing and characterisation.
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- 2020
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11. Corrigendum to 'Generation of a set of isogenic, gene-edited iPSC lines homozygous for all main APOE variants and an APOE knock-out line' [Stem Cell Res. 34/1873–5061 (2019) 101349–55]
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Benjamin Schmid, Kennie R. Prehn, Natakarn Nimsanor, Blanca Irene Aldana Garcia, Ulla Poulsen, Ida Jørring, Mikkel A. Rasmussen, Christian Clausen, Ulrike A. Mau-Holzmann, Sarayu Ramakrishna, Ravi Muddashetty, Rachel Steeg, Kevin Bruce, Peter Mackintosh, Andreas Ebneth, Bjørn Holst, and Alfredo Cabrera-Socorro
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Biology (General) ,QH301-705.5 - Published
- 2020
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12. Deploying human pluripotent stem cells to treat central nervous system disorders: facts, challenges and realising the potential
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Timothy E Allsopp, Andreas Ebneth, and Alfredo Cabrera-Socorro
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Biology (General) ,QH301-705.5 - Abstract
Human pluripotent stem cells (hPSC) represent a unique opportunity to study fundamental biological processes in a human- and cell-specific setting. Its translational potential and the impact on human health makes this technology revolutionary. The possibility to generate stem cells from almost any somatic cell, and their capacity to be differentiated in virtually all cells of the body has been demonstrated extensively during the last decade of research. Target-centric as well as phenotypic screenings using differentiated cells have become a reality, while the use of these cells for “disease modelling” is still challenging due to the paucity of relevant and reproducible phenotypes. The combination of hPSCs with gene editing technologies aiming to e.g. reduce immunogenic response has enabled promising clinical trials that will eventually demonstrate their therapeutic potential in tissue regeneration and cancer treatment. Maximizing the therapeutic applications of hPSCs requires systematic data comparison, consensus between scientists and health care professionals, as well as a close collaboration between research labs, clinics, and regulators. The goal of this review is to provide a comprehensive outlook of the current use of hPSCs in drug development and regenerative medicine for the treatment of central nervous system (CNS) disorders. In the first part, we analyse how hPSCs are currently used in drug development and discuss their use in challenging paradigms such as neurodegeneration. In the second part we review the status of hPSCs in regenerative medicine. Finally, key challenges and pitfalls of the technology will be discussed, and strategies proposed to improve hPSC research and to benefit patients across different therapeutic areas.
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- 2019
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13. Genetically Engineered iPSC-Derived FTDP-17 MAPT Neurons Display Mutation-Specific Neurodegenerative and Neurodevelopmental Phenotypes
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An Verheyen, Annick Diels, Joke Reumers, Kirsten Van Hoorde, Ilse Van den Wyngaert, Constantin van Outryve d’Ydewalle, An De Bondt, Jacobine Kuijlaars, Louis De Muynck, Ronald De Hoogt, Alexis Bretteville, Steffen Jaensch, Arjan Buist, Alfredo Cabrera-Socorro, Selina Wray, Andreas Ebneth, Peter Roevens, Ines Royaux, and Pieter J. Peeters
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Medicine (General) ,R5-920 ,Biology (General) ,QH301-705.5 - Abstract
Summary: Tauopathies such as frontotemporal dementia (FTD) remain incurable to date, partially due to the lack of translational in vitro disease models. The MAPT gene, encoding the microtubule-associated protein tau, has been shown to play an important role in FTD pathogenesis. Therefore, we used zinc finger nucleases to introduce two MAPT mutations into healthy donor induced pluripotent stem cells (iPSCs). The IVS10+16 mutation increases the expression of 4R tau, while the P301S mutation is pro-aggregant. Whole-transcriptome analysis of MAPT IVS10+16 neurons reveals neuronal subtype differences, reduced neural progenitor proliferation potential, and aberrant WNT/SHH signaling. Notably, these neurodevelopmental phenotypes could be recapitulated in neurons from patients carrying the MAPT IVS10+16 mutation. Moreover, the additional pro-aggregant P301S mutation revealed additional phenotypes, such as an increased calcium burst frequency, reduced lysosomal acidity, tau oligomerization, and neurodegeneration. This series of iPSCs could serve as a platform to unravel a potential link between pathogenic 4R tau and FTD. : In this article, Verheyen and colleagues introduced 2 MAPT mutations (IVS10+16/P301S) into healthy donor iPSCs. Characterization of MAPT IVS10+16 neurons revealed neurodevelopmental neuron subtype differences, reduced neural progenitor proliferation, and aberrant WNT/SHH signaling, while the additional pro-aggregant P301S mutation revealed phenotypes more related to neurodegeneration. These iPSCs could help to unravel a potential link between pathogenic 4R tau and FTD. Keywords: FTDP-17, MAPT, iPSC, ZFN, disease modelling
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- 2018
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14. Overexpression of Tau Protein Inhibits Kinesin-Dependent Trafficking of Vesicles, Mitochondria, and Endoplasmic Reticulum: Implications for Alzheimer's Disease
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Ebneth, A., Godemann, R., Stamer, K., Illenberger, S., Trinczek, B., and Mandelkow, E.
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- 1998
15. Rapid establishment of the European Bank for induced Pluripotent Stem Cells (EBiSC) - the Hot Start experience
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Paul A. De Sousa, Rachel Steeg, Elisabeth Wachter, Kevin Bruce, Jason King, Marieke Hoeve, Shalinee Khadun, George McConnachie, Julie Holder, Andreas Kurtz, Stefanie Seltmann, Johannes Dewender, Sascha Reimann, Glyn Stacey, Orla O'Shea, Charlotte Chapman, Lyn Healy, Heiko Zimmermann, Bryan Bolton, Trisha Rawat, Isobel Atkin, Anna Veiga, Bernd Kuebler, Blanca Miranda Serano, Tomo Saric, Jürgen Hescheler, Oliver Brüstle, Michael Peitz, Cornelia Thiele, Niels Geijsen, Bjørn Holst, Christian Clausen, Majlinda Lako, Lyle Armstrong, Shailesh K. Gupta, Alexander J. Kvist, Ryan Hicks, Anna Jonebring, Gabriella Brolén, Andreas Ebneth, Alfredo Cabrera-Socorro, Patrik Foerch, Martine Geraerts, Tina C. Stummann, Shawn Harmon, Carol George, Ian Streeter, Laura Clarke, Helen Parkinson, Peter W. Harrison, Adam Faulconbridge, Luca Cherubin, Tony Burdett, Cesar Trigueros, Minal J Patel, Christa Lucas, Barry Hardy, Rok Predan, Joh Dokler, Maja Brajnik, Oliver Keminer, Ole Pless, Philip Gribbon, Carsten Claussen, Annette Ringwald, Beate Kreisel, Aidan Courtney, and Timothy E. Allsopp
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Biology (General) ,QH301-705.5 - Abstract
A fast track “Hot Start” process was implemented to launch the European Bank for Induced Pluripotent Stem Cells (EBiSC) to provide early release of a range of established control and disease linked human induced pluripotent stem cell (hiPSC) lines. Established practice amongst consortium members was surveyed to arrive at harmonised and publically accessible Standard Operations Procedures (SOPs) for tissue procurement, bio-sample tracking, iPSC expansion, cryopreservation, qualification and distribution to the research community. These were implemented to create a quality managed foundational collection of lines and associated data made available for distribution. Here we report on the successful outcome of this experience and work flow for banking and facilitating access to an otherwise disparate European resource, with lessons to benefit the international research community. eTOC: The report focuses on the EBiSC experience of rapidly establishing an operational capacity to procure, bank and distribute a foundational collection of established hiPSC lines. It validates the feasibility and defines the challenges of harnessing and integrating the capability and productivity of centres across Europe using commonly available resources currently in the field.
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- 2017
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16. Generation of a set of isogenic, gene-edited iPSC lines homozygous for all main APOE variants and an APOE knock-out line
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Benjamin Schmid, Kennie R. Prehn, Natakarn Nimsanor, Blanca Irene Aldana Garcia, Ulla Poulsen, Ida Jørring, Mikkel A. Rasmussen, Christian Clausen, Ulrike A. Mau-Holzmann, Sarayu Ramakrishna, Ravi Muddashetty, Rachel Steeg, Kevin Bruce, Peter Mackintosh, Andreas Ebneth, Bjørn Holst, and Alfredo Cabrera-Socorro
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Biology (General) ,QH301-705.5 - Abstract
Alzheimer's disease (AD) is the most frequent neurodegenerative disease amongst the elderly. The SNPs rs429358 and rs7412 in the APOE gene are the most common risk factor for sporadic AD, and there are three different alleles commonly referred to as APOE-ε2, APOE-ε3 and APOE-ε4. Induced pluripotent stem cells (iPSCs) hold great promise to model AD as such cells can be differentiated in vitro to the required cell type. Here we report the use of CRISPR/Cas9 technology employed on iPSCs from a healthy individual with an APOE-ε3/ε4 genotype to obtain isogenic APOE-ε2/ε2, APOE-ε3/ε3, APOE-ε4/ε4 lines as well as an APOE-knock-out line.
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- 2019
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17. Development of a Scalable, High-Throughput-Compatible Assay to Detect Tau Aggregates Using iPSC-Derived Cortical Neurons Maintained in a Three-Dimensional Culture Format
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Medda, X., Mertens, L., Versweyveld, S., Diels, A., Barnham, L., Bretteville, A., Buist, A., Verheyen, A., Royaux, I., Ebneth, A., and Cabrera-Socorro, A.
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- 2016
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18. Pharmacological characterization of mutant huntingtin aggregate-directed PET imaging tracer candidates
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Madlen Hotze, Miriam Gehrmann, Manuela Hessmann, Longbin Liu, Christoph Scheich, Kathrin Petersen, Nicole Hoeschen, Peter Johnson, Sabine Schaertl, Stefanie Jahn, Matthew R. Mills, Ignacio Munoz-Sanjuan, Alex S. Kiselyov, John Wityak, Anass Chiki, Vinod Khetarpal, Karsten Kottig, Gerhard Schmiedel, Galina Bursow, Stefanie Ladewig, Christopher J. Brown, Karola Berg-Rosseburg, Michael Prime, Sven Letschert, Andreas Ebneth, Jonathan Bard, Celia Dominguez, Frank Herrmann, and Hilal A. Lashuel
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exon-1 ,Huntingtin ,Mutant ,law.invention ,Mice ,Radioligand Assay ,amyloid fibrils ,Exon ,law ,alzheimers-disease ,Huntingtin Protein ,dysfunction ,Multidisciplinary ,Chemistry ,Immunohistochemistry ,Recombinant Proteins ,Cell biology ,small molecules ,Huntington Disease ,Recombinant DNA ,Medicine ,Preclinical imaging ,congenital, hereditary, and neonatal diseases and abnormalities ,brain ,Science ,Mice, Transgenic ,Protein Aggregation, Pathological ,Article ,Protein Aggregates ,Alzheimer Disease ,mental disorders ,Animals ,Humans ,Radioactive Tracers ,beta-sheet structure ,Pharmacology ,Nitrogen Radioisotopes ,toxicity ,In vitro ,nervous system diseases ,Disease Models, Animal ,polyglutamine aggregation ,Positron-Emission Tomography ,Diseases of the nervous system ,Autoradiography ,identification ,Radiopharmaceuticals ,Trinucleotide repeat expansion ,Biomarkers ,Ex vivo - Abstract
Huntington’s disease (HD) is caused by a CAG trinucleotide repeat expansion in the first exon of the huntingtin (HTT) gene coding for the huntingtin (HTT) protein. The misfolding and consequential aggregation of CAG-expanded mutant HTT (mHTT) underpin HD pathology. Our interest in the life cycle of HTT led us to consider the development of high-affinity small-molecule binders of HTT oligomerized/amyloid-containing species that could serve as either cellular and in vivo imaging tools or potential therapeutic agents. We recently reported the development of PET tracers CHDI-180 and CHDI-626 as suitable for imaging mHTT aggregates, and here we present an in-depth pharmacological investigation of their binding characteristics. We have implemented an array of in vitro and ex vivo radiometric binding assays using recombinant HTT, brain homogenate-derived HTT aggregates, and brain sections from mouse HD models and humans post-mortem to investigate binding affinities and selectivity against other pathological proteins from indications such as Alzheimer’s disease and spinocerebellar ataxia 1. Radioligand binding assays and autoradiography studies using brain homogenates and tissue sections from HD mouse models showed that CHDI-180 and CHDI-626 specifically bind mHTT aggregates that accumulate with age and disease progression. Finally, we characterized CHDI-180 and CHDI-626 regarding their off-target selectivity and binding affinity to beta amyloid plaques in brain sections and homogenates from Alzheimer’s disease patients.
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- 2021
19. Scalable expansion of iPSC and their derivatives across multiple lineages
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Chee Keong Kwok, Isabelle Sébastien, Krithika Hariharan, Ina Meiser, Jeanette Wihan, Saskia Altmaier, Isabell Karnatz, Dominic Bauer, Benjamin Fischer, Alexander Feile, Alfredo Cabrera-Socorro, Mikkel Rasmussen, Bjørn Holst, Julia C. Neubauer, Christian Clausen, Catherine Verfaillie, Andreas Ebneth, Mattias Hansson, Rachel Steeg, Heiko Zimmermann, and Publica
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Pluripotent Stem Cells ,upscaling ,induced pluripotent stem cells ,bioprocessing ,Induced Pluripotent Stem Cells ,suspension-based bioreactor ,Cell Culture Techniques ,Cell Differentiation ,EBiSC ,differentiation ,cell processing ,Toxicology ,Bioreactors ,cell banking - Abstract
Induced pluripotent stem cell (iPSC) technology enabled the production of pluripotent stem cell lines from somatic cells from a range of known genetic backgrounds. Their ability to differentiate and generate a wide variety of cell types has resulted in their use for various biomedical applications, including toxicity testing. Many of these iPSC lines are now registered in databases and stored in biobanks such as the European Bank for induced pluripotent Stem Cells (EBiSC), which can streamline the quality control and distribution of these individual lines. To generate the quantities of cells for banking and applications like high-throughput toxicity screening, scalable and robust methods need to be developed to enable the large-scale production of iPSCs. 3D suspension culture platforms are increasingly being used by stem cell researchers, owing to a higher cell output in a smaller footprint, as well as simpler scaling by increasing culture volume. Here we describe our strategies for successful scalable production of iPSCs using a benchtop bioreactor and incubator for 3D suspension cultures, while maintaining quality attributes expected of high-quality iPSC lines. Additionally, to meet the increasing demand for "ready-to-use" cell types, we report recent work to establish robust, scalable differentiation protocols to cardiac, neural, and hepatic fate to enable EBiSC to increase available research tools. ispartof: REPRODUCTIVE TOXICOLOGY vol:112 pages:23-35 ispartof: location:United States status: published
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- 2022
20. Development of LC/MS/MS, High-Throughput Enzymatic and Cellular Assays for the Characterization of Compounds That Inhibit Kynurenine Monooxygenase (KMO)
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Winkler, Dirk, Beconi, Maria, Toledo-Sherman, Leticia M., Prime, Michael, Ebneth, Andreas, Dominguez, Celia, and Muñoz-Sanjuan, Ignacio
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- 2013
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21. Generation of three isogenic, gene-edited iPSC lines carrying the APOE-Christchurch mutation into the three common APOE variants: APOE2Ch, APOE3Ch and APOE4Ch
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Haidar, Mansour, Schmid, Benjamin, Ruiz, Agustín, Ebneth, Andreas, and Cabrera-Socorro, Alfredo
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- 2024
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22. Characterization of HTT inclusion size, location, and timing in the zQ175 mouse model of Huntington's disease: an in vivo high-content imaging study.
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Nikisha Carty, Nadège Berson, Karsten Tillack, Christina Thiede, Diana Scholz, Karsten Kottig, Yalda Sedaghat, Christina Gabrysiak, George Yohrling, Heinz von der Kammer, Andreas Ebneth, Volker Mack, Ignacio Munoz-Sanjuan, and Seung Kwak
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Medicine ,Science - Abstract
Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder caused by a CAG trinucleotide repeat expansion in the huntingtin gene. Major pathological hallmarks of HD include inclusions of mutant huntingtin (mHTT) protein, loss of neurons predominantly in the caudate nucleus, and atrophy of multiple brain regions. However, the early sequence of histological events that manifest in region- and cell-specific manner has not been well characterized. Here we use a high-content histological approach to precisely monitor changes in HTT expression and characterize deposition dynamics of mHTT protein inclusion bodies in the recently characterized zQ175 knock-in mouse line. We carried out an automated multi-parameter quantitative analysis of individual cortical and striatal cells in tissue slices from mice aged 2-12 months and confirmed biochemical reports of an age-associated increase in mHTT inclusions in this model. We also found distinct regional and subregional dynamics for inclusion number, size and distribution with subcellular resolution. We used viral-mediated suppression of total HTT in the striatum of zQ175 mice as an example of a therapeutically-relevant but heterogeneously transducing strategy to demonstrate successful application of this platform to quantitatively assess target engagement and outcome on a cellular basis.
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- 2015
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23. Development of a fully human assay combining NGN2-inducible neurons co-cultured with iPSC-derived astrocytes amenable for electrophysiological studies
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Shih, Pei-Yu, Kreir, Mohamed, Kumar, Devesh, Seibt, Frederik, Pestana, Francisco, Schmid, Benjamin, Holst, Bjørn, Clausen, Christian, Steeg, Rachel, Fischer, Benjamin, Pita-Almenar, Juan, Ebneth, Andreas, and Cabrera-Socorro, Alfredo
- Published
- 2021
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24. SOX9-induced Generation of Functional Astrocytes Supporting Neuronal Maturation in an All-human System
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Alfredo Cabrera-Socorro, Pei-Yu Shih, Juan Diego Pita Almenar, Jonathan De Smedt, Johanna Van Daele, Devesh Kumar, Tim Vervliet, Katrien Neyrinck, Astrid D'hondt, Melissa Nijs, Catherine M. Verfaillie, Mohamed Kreir, Geert Bultynck, Keimpe D. Wierda, Mélanie Planque, Tom Vanbokhoven, Andreas Ebneth, Vania Broccoli, Frederik Seibt, and Sarah-Maria Fendt
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Genome engineering ,Cell type ,medicine.medical_treatment ,Neurogenesis ,Induced Pluripotent Stem Cells ,SOX9 ,Biology ,Article ,All-human co-culture system ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Neural Stem Cells ,Pluripotent stem cells ,medicine ,Animals ,Humans ,Secretion ,Induced pluripotent stem cell ,030304 developmental biology ,Neurons ,0303 health sciences ,Growth factor ,SOX9 Transcription Factor ,General Medicine ,Differentiation protocol ,Neural stem cell ,Cytokine ,Astrocytes ,Stem cell ,Neuroscience ,030217 neurology & neurosurgery - 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. Graphical Abstract
- Published
- 2021
25. Pharmacological characterization of mutant huntingtin aggregate-directed PET imaging tracer candidates
- Author
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Herrmann, Frank, primary, Hessmann, Manuela, additional, Schaertl, Sabine, additional, Berg-Rosseburg, Karola, additional, Brown, Christopher J, additional, Bursow, Galina, additional, Chiki, Anass, additional, Ebneth, Andreas, additional, Gehrmann, Miriam, additional, Hoeschen, Nicole, additional, Hotze, Madlen, additional, Jahn, Stefanie, additional, Johnson, Peter D, additional, Khetarpal, Vinod, additional, Kiselyov, Alex, additional, Kottig, Karsten, additional, Ladewig, Stefanie, additional, Lashuel, Hilal, additional, Letschert, Sven, additional, Mills, Matthew R, additional, Petersen, Kathrin, additional, Prime, Michael E, additional, Scheich, Christoph, additional, Schmiedel, Gerhard, additional, Wityak, John, additional, Liu, Longbin, additional, Dominguez, Celia, additional, Muñoz-Sanjuán, Ignacio, additional, and Bard, Jonathan A, additional
- Published
- 2021
- Full Text
- View/download PDF
26. Small Molecule Binding to Alzheimer Risk Factor CD33 Promotes Aβ Phagocytosis
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Andreas Ebneth, Luke A. Miles, S.J. Hermans, Marie-Laure Rives, Daniel Oehlrich, Berthold Wroblowski, Jasmina Markulić, Michael W. Parker, Ines Royaux, Nancy C. Hancock, Gabriela A. N. Crespi, Andrés A. Trabanco, Jonathan H. Gooi, Larissa Doughty, and Tracy L. Nero
- Subjects
0301 basic medicine ,Peptide ,02 engineering and technology ,Molecular neuroscience ,Alzheimer's Disease ,Article ,03 medical and health sciences ,chemistry.chemical_compound ,medicine ,Receptor ,lcsh:Science ,Components of the Immune System ,chemistry.chemical_classification ,Multidisciplinary ,Molecular Structure ,Microglia ,021001 nanoscience & nanotechnology ,Protein Structure Aspects ,3. Good health ,Sialic acid ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,chemistry ,lcsh:Q ,CD33 ,Molecular Neuroscience ,Small molecule binding ,Pharmacophore ,0210 nano-technology ,Glycoprotein ,Neuroscience - Abstract
Summary Polymorphism in the microglial receptor CD33 gene has been linked to late-onset Alzheimer disease (AD), and reduced expression of the CD33 sialic acid-binding domain confers protection. Thus, CD33 inhibition might be an effective therapy against disease progression. Progress toward discovery of selective CD33 inhibitors has been hampered by the absence of an atomic resolution structure. We report here the crystal structures of CD33 alone and bound to a subtype-selective sialic acid mimetic called P22 and use them to identify key binding residues by site-directed mutagenesis and binding assays to reveal the molecular basis for its selectivity toward sialylated glycoproteins and glycolipids. We show that P22, when presented on microparticles, increases uptake of the toxic AD peptide, amyloid-β (Aβ), into microglial cells. Thus, the sialic acid-binding site on CD33 is a promising pharmacophore for developing therapeutics that promote clearance of the Aβ peptide that is thought to cause AD., Graphical Abstract, Highlights • A sialic acid mimetic increases the uptake of Alzheimer peptide into microglia • Crystal structure of cell surface receptor CD33 bound to the sialic acid mimetic • Crystal structure of unliganded CD33 and mutagenesis studies revealed key residues • CD33 carbohydrate-binding site is a pharmacophore for Alzheimer drug development, Molecular Structure; Neuroscience; Molecular Neuroscience; Components of the Immune System; Protein Structure Aspects
- Published
- 2019
27. General Roles of Abscisic and Jasmonic Acids in Gene Activation as a Result of Mechanical Wounding
- Author
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Hildmann, Thomas, Ebneth, Marcus, Peña-Cortés, Hugo, Sánchez-Serrano, Jose J., Willmitzer, Lothar, and Prat, Salomé
- Published
- 1992
- Full Text
- View/download PDF
28. Generation of a set of isogenic iPSC lines carrying all APOE genetic variants (Ɛ2/Ɛ3/Ɛ4) and knock-out for the study of APOE biology in health and disease
- Author
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Christian Clausen, Alfredo Ramirez, Andreas Ebneth, Pamela V. Martino-Adami, Alfredo Cabrera-Socorro, Eva Díaz-Guerra, Oliver Brüstle, Michaela Thoenes, Benjamin Schmid, Peter Reinhardt, Clara Grezella, Lamiaa Bahnassawy, Carlos Vicario, Bjørn Holst, Margot H.M. Bakker, Michael Peitz, Klaus Fliessbach, Innovative Medicines Initiative, and European Commission
- Subjects
0301 basic medicine ,Apolipoprotein E ,Apolipoprotein B ,Genotype ,Induced Pluripotent Stem Cells ,genetics [Alzheimer Disease] ,Disease ,Homology (biology) ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Apolipoproteins E ,Alzheimer Disease ,ddc:570 ,Animals ,Induced pluripotent stem cell ,Gene ,lcsh:QH301-705.5 ,Biology ,Genetics ,biology ,Cell Biology ,General Medicine ,In vitro ,030104 developmental biology ,lcsh:Biology (General) ,biology.protein ,genetics [Apolipoproteins E] ,lipids (amino acids, peptides, and proteins) ,030217 neurology & neurosurgery ,Developmental Biology - Abstract
APOE genotype is the strongest genetic risk factor for Alzheimer’s Disease (AD). The low degree of homology between mouse and human APOE is a concerning issue in preclinical models currently used to study the role of this gene in AD pathophysiology. A key objective of ADAPTED (Alzheimer’s Disease Apolipoprotein Pathology for Treatment Elucidation and Development) project was to generate in vitro models that better recapitulate human APOE biology. We describe a new set of induced pluripotent stem cells (iPSC) lines carrying common APOE variants (Ɛ2, Ɛ3, and Ɛ3/Ɛ4) and a knock-out isogenic to the parental APOE Ɛ4/Ɛ4 line (UKBi011-A)., This study was funded by the ADAPTED (Alzheimer’s Disease Apolipoprotein Pathology for Treatment Elucidation and Development) consortium which has received funding from the Innovative Medicines Initiative 2 Joint Undertaking under Grant Agreement No 115975. This Joint Undertaking receives support from the European Union’s Horizon 2020 research and innovation programme and the European Federation of Pharmaceutical Industries and Associations.
- Published
- 2021
29. Generation of a set of isogenic iPSC lines carrying all APOE genetic variants (epsilon 2/epsilon 3/epsilon 4) and knock-out for the study of APOE biology in health and disease
- Author
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Schmid, Benjamin, Holst, Bjorn, Clausen, Christian, Bahnassawy, Lamiaa, Reinhardt, Peter, Bakker, Margot H. M., Diaz-Guerra, Eva, Vicario, Carlos, Martino-Adami, Pamela, V, Thoenes, Michaela, Ramirez, Alfredo, Flissbach, Klaus, Grezella, Clara, Bruestle, Oliver, Peitz, Michael, Ebneth, Andreas, Cabrera-Socorro, Alfredo, Schmid, Benjamin, Holst, Bjorn, Clausen, Christian, Bahnassawy, Lamiaa, Reinhardt, Peter, Bakker, Margot H. M., Diaz-Guerra, Eva, Vicario, Carlos, Martino-Adami, Pamela, V, Thoenes, Michaela, Ramirez, Alfredo, Flissbach, Klaus, Grezella, Clara, Bruestle, Oliver, Peitz, Michael, Ebneth, Andreas, and Cabrera-Socorro, Alfredo
- Abstract
APOE genotype is the strongest genetic risk factor for Alzheimer's Disease (AD). The low degree of homology between mouse and human APOE is a concerning issue in preclinical models currently used to study the role of this gene in AD pathophysiology. A key objective of ADAPTED (Alzheimer's Disease Apolipoprotein Pathology for Treatment Elucidation and Development) project was to generate in vitro models that better recapitulate human APOE biology. We describe a new set of induced pluripotent stem cells (iPSC) lines carrying common APOE variants (epsilon 2,epsilon 3, and epsilon 3/epsilon 4) and a knock-out isogenic to the parental APOE epsilon 4/epsilon 4 line (UKBi011-A).
- Published
- 2021
30. Generation of two gene edited iPSC-lines carrying a DOX-inducible NGN2 expression cassette with and without GFP in the AAVS1 locus
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Alfredo Cabrera-Socorro, Benjamin Schmid, Ida Jørring, Andreas Ebneth, Bjørn Holst, Ulrike A. Mau-Holzmann, Ulla B. Poulsen, Rachel Steeg, Mikkel A. Rasmussen, Christian Clausen, Hugh Nuthall, and Publica
- Subjects
0301 basic medicine ,Gene Editing ,Transgene ,Cellular differentiation ,Induced Pluripotent Stem Cells ,Cell Differentiation ,Cell Biology ,General Medicine ,Biology ,Green fluorescent protein ,Cell biology ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,lcsh:Biology (General) ,Genes, Reporter ,Expression cassette ,Transgenes ,Stem cell ,Induced pluripotent stem cell ,Transcription factor ,Gene ,lcsh:QH301-705.5 ,030217 neurology & neurosurgery ,Developmental Biology - Abstract
Neurog2 is the gene encoding the neuronal transcription factor NGN2, which can convert stem cells into functional neurons in a fast and efficient way. Here we report the generation of two iPS cell lines, where DOX inducible constructs of neurog2 either without or with T2A-eGFP were inserted into the safe-site locus AAVS1. These iPS cell lines, BIONi010-C-13 and BIONi010-C-15, respectively, stay pluripotent without DOX but differentiate to (GFP positive) neurons when DOX is added without the need of differentiation factors.
- Published
- 2020
31. Corrigendum to “Generation of a set of isogenic, gene-edited iPSC lines homozygous for all main APOE variants and an APOE knock-out line” [Stem Cell Res. 34/1873–5061 (2019) 101349–55]
- Author
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Schmid, Benjamin, Prehn, Kennie R., Nimsanor, Natakarn, Garcia, Blanca Irene Aldana, Poulsen, Ulla, Jørring, Ida, Rasmussen, Mikkel A., Clausen, Christian, Mau-Holzmann, Ulrike A., Ramakrishna, Sarayu, Muddashetty, Ravi, Steeg, Rachel, Bruce, Kevin, Mackintosh, Peter, Ebneth, Andreas, Holst, Bjørn, and Cabrera-Socorro, Alfredo
- Published
- 2020
- Full Text
- View/download PDF
32. Generation of a set of isogenic iPSC lines carrying all APOE genetic variants (Ɛ2/Ɛ3/Ɛ4) and knock-out for the study of APOE biology in health and disease
- Author
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Schmid, Benjamin, primary, Holst, Bjørn, additional, Clausen, Christian, additional, Bahnassawy, Lamiaa, additional, Reinhardt, Peter, additional, Bakker, Margot H.M., additional, Díaz-Guerra, Eva, additional, Vicario, Carlos, additional, Martino-Adami, Pamela V., additional, Thoenes, Michaela, additional, Ramirez, Alfredo, additional, Fliessbach, Klaus, additional, Grezella, Clara, additional, Brüstle, Oliver, additional, Peitz, Michael, additional, Ebneth, Andreas, additional, and Cabrera-Socorro, Alfredo, additional
- Published
- 2021
- Full Text
- View/download PDF
33. Generation of two gene edited iPSC-lines carrying a DOX-inducible NGN2 expression cassette with and without GFP in the AAVS1 locus
- Author
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Schmid, Benjamin, primary, Holst, Bjørn, additional, Poulsen, Ulla, additional, Jørring, Ida, additional, Clausen, Christian, additional, Rasmussen, M., additional, Mau-Holzmann, Ulrike A., additional, Steeg, Rachel, additional, Nuthall, Hugh, additional, Ebneth, A., additional, and Cabrera-Socorro, A., additional
- Published
- 2021
- Full Text
- View/download PDF
34. Mittelalter und Mediävistik in NDB und Deutscher Biographie
- Author
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Ebneth, Bernhard and Reinert, Matthias
- Abstract
Die Deutsche Biographie - historisch-biographische Infomationen und das Nutzungspotential für die mediävistische Forschung: Schnittstellen, Normdaten, Kooperation am Beispiel der Metadaten der Biographien von Kaiserinnen und Königinnen des Mittelalters.
- Published
- 2020
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- View/download PDF
35. Generation of a human induced pluripotent stem cell–based model for tauopathies combining three microtubule‐associated protein TAU mutations which displays several phenotypes linked to neurodegeneration
- Author
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Joke Terryn, Keimpe D. Wierda, Bart De Strooper, Alfredo Cabrera-Socorro, Frederic Lluis, Francisco Pestana, Andreas Ebneth, Laura Ordovás, Juan Antonio García-León, Ana Quiles, Catherine M. Verfaillie, Ann Swijsen, Fatemeharefeh Nami, Lutgarde Serneels, Kristel Eggermont, Raheem Fazal, Mohamed Kreir, Lieven Thorrez, Annerieke Sierksma, and Philip Van Damme
- Subjects
0301 basic medicine ,Epidemiology ,Membrane Potentials ,CRISPR ,CRISPR-Cas ,Induced pluripotent stem cell ,PIGGYBAC TRANSPOSON ,Neurons ,Health Policy ,MISSENSE ,Neurodegeneration ,CORTICAL-NEURONS ,MOUSE MODEL ,Alzheimer's disease ,Phenotype ,NEOCORTEX ,Cell biology ,ALZHEIMERS-DISEASE ,Psychiatry and Mental health ,Disease modeling ,Drug screening ,Tauopathies ,Tauopathy ,Life Sciences & Biomedicine ,Frontotemporal dementia ,Neurite ,Neurogenesis ,Induced Pluripotent Stem Cells ,Neuronal Outgrowth ,Clinical Neurology ,INHIBITION ,Parkinsonism linked to chromosome 17 ,tau Proteins ,Biology ,Cell Line ,Progressive supranuclear palsy ,03 medical and health sciences ,Cellular and Molecular Neuroscience ,Developmental Neuroscience ,Downregulation and upregulation ,medicine ,Humans ,Science & Technology ,medicine.disease ,PATHOLOGY ,030104 developmental biology ,Mutation ,Nerve Degeneration ,HIPPOCAMPUS ,Neurosciences & Neurology ,Neurology (clinical) ,CRISPR-Cas Systems ,Geriatrics and Gerontology ,Transcriptome - Abstract
INTRODUCTION: Tauopathies are neurodegenerative diseases characterized by TAU protein-related pathology, including frontotemporal dementia and Alzheimer's disease among others. Mutant TAU animal models are available, but none of them faithfully recapitulates human pathology and are not suitable for drug screening. METHODS: To create a new in vitro tauopathy model, we generated a footprint-free triple MAPT-mutant human induced pluripotent stem cell line (N279K, P301L, and E10+16 mutations) using clustered regularly interspaced short palindromic repeats-FokI and piggyBac transposase technology. RESULTS: Mutant neurons expressed pathogenic 4R and phosphorylated TAU, endogenously triggered TAU aggregation, and had increased electrophysiological activity. TAU-mutant cells presented deficiencies in neurite outgrowth, aberrant sequence of differentiation to cortical neurons, and a significant activation of stress response pathways. RNA sequencing confirmed stress activation, demonstrated a shift toward GABAergic identity, and an upregulation of neurodegenerative pathways. DISCUSSION: In summary, we generated a novel in vitro human induced pluripotent stem cell TAU-mutant model displaying neurodegenerative disease phenotypes that could be used for disease modeling and drug screening. ispartof: ALZHEIMERS & DEMENTIA vol:14 issue:10 pages:1261-1280 ispartof: location:United States status: published
- Published
- 2018
36. Evaluation of a high-throughput fluorescence assay method for hERG potassium channel inhibition
- Author
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Dorn, Arnulf, Hermann, Francis, Ebneth, Andreas, Bothmann, Hendrick, Trube, Gerhard, Christensen, Klaus, and Apfel, Christian
- Subjects
Electrophysiology -- Analysis -- Research ,Potassium channels -- Research -- Analysis ,Fluorescence -- Analysis -- Research ,Biological sciences ,Analysis ,Research - Abstract
The number of projects in drug development that fail in late phases because of cardiac side effects such as QT prolongation can impede drug discovery and development of projects. The [...]
- Published
- 2005
37. Identification and functional characterization of a novel KCNE2 (MiRP1) mutation that alters HERG channel kinetics
- Author
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Isbrandt, Dirk, Friederich, Patrick, Solth, Anna, Haverkamp, Wilhelm, Ebneth, Andreas, Borggrefe, Martin, Funke, Harald, Sauter, Kathrin, Breithardt, Günter, Pongs, Olaf, and Schulze-Bahr, Eric
- Published
- 2002
- Full Text
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38. Development of a fully human assay combining NGN2-inducible neurons co-cultured with iPSC-derived astrocytes amenable for electrophysiological studies
- Author
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Benjamin Schmid, Pei-Yu Shih, Devesh Kumar, Alfredo Cabrera-Socorro, Christian Clausen, Andreas Ebneth, Francisco Pestana, Rachel Steeg, Bjørn Holst, Frederik Seibt, Juan D. Pita-Almenar, Mohamed Kreir, Benjamin Fischer, and Publica
- Subjects
QH301-705.5 ,Somatic cell ,Cellular differentiation ,Induced Pluripotent Stem Cells ,Biology ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Cell & Tissue Engineering ,Basic Helix-Loop-Helix Transcription Factors ,Animals ,Humans ,Neurogenin-2 ,Biology (General) ,Progenitor cell ,Induced pluripotent stem cell ,Cells, Cultured ,030304 developmental biology ,Neurons ,0303 health sciences ,Science & Technology ,Cell Differentiation ,Cell Biology ,General Medicine ,Phenotype ,Embryonic stem cell ,Coculture Techniques ,Cell biology ,SYNAPTIC-TRANSMISSION ,DIFFERENTIATION ,Biotechnology & Applied Microbiology ,Astrocytes ,EXCITATORY NEURONS ,Stem cell ,Life Sciences & Biomedicine ,PLURIPOTENT STEM-CELLS ,030217 neurology & neurosurgery ,Developmental Biology - Abstract
Neurogenin 2 encodes a neural-specific transcription factor (NGN2) able to drive neuronal fate on somatic and stem cells. NGN2 is expressed in neural progenitors within the developing central and peripheral nervous systems. Overexpression of NGN2 in human induced pluripotent stem cells (hiPSCs) or human embryonic stem cells has been shown to efficiently trigger conversion to neurons. Here we describe two gene-edited hiPSC lines harbouring a doxycycline (DOX)-inducible cassette in the AAVS1 locus driving expression of NGN2 (BIONi010-C-13) or NGN2-T2A-GFP (BIONi010-C-15). By introducing NGN2-expressing cassette, we reduce variability associated with conventional over-expression methods such as viral transduction, making these lines amenable for scale-up production and screening processes. DOX-treated hiPSCs convert to neural phenotype within one week and display the expression of structural neuronal markers such as Beta-III tubulin and tau. We performed functional characterization of NGN2-neurons co-cultured with hiPSC-derived astrocytes in a "fully-humanized" set up. Passive properties of NGN2-neurons were indistinguishable from mouse primary cells while displaying variable activity in extracellular recordings performed in multi-electrode arrays (MEAs). We demonstrate that hiPSC-derived astrocytes and neurons can be co-cultured and display functional properties comparable to the gold standard used in electrophysiology. Both lines are globally available via EBiSC repository at https://cells.ebisc.org/. ispartof: STEM CELL RESEARCH vol:54 ispartof: location:England status: published
- Published
- 2021
39. Rapid establishment of the European Bank for induced Pluripotent Stem Cells (EBiSC) - the Hot Start experience
- Author
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Oliver Brüstle, Annette Ringwald, Christian Clausen, Charlotte Chapman, Glyn Stacey, Alexander J. Kvist, Marieke A. Hoeve, Shailesh Kumar Gupta, Blanca Miranda Serano, Minal Patel, Gabriella Brolén, Tomo Saric, George McConnachie, Cornelia Thiele, Julie Holder, Trisha Rawat, Tina C. Stummann, Joh Dokler, Maja Brajnik, Bryan Bolton, Lyle Armstrong, Bernd Kuebler, Alfredo Cabrera-Socorro, Isobel Atkin, Adam Faulconbridge, Christa Lucas, Jason A. King, Johannes Dewender, Ian Streeter, Rok Predan, Majlinda Lako, Lyn Healy, Bjørn Holst, Martine Geraerts, Luca Cherubin, Carsten Claussen, Heiko Zimmermann, Timothy E Allsopp, Orla O'Shea, Peter W. Harrison, K. Bruce, Niels Geijsen, Michael Peitz, Tony Burdett, Paul A. De Sousa, Ole Pless, Elisabeth Wachter, Anna Veiga, A. Courtney, Laura Clarke, Helen Parkinson, Cesar Trigueros, Anna Jonebring, Shalinee Khadun, Rachel Steeg, Patrik Foerch, Sascha Reimann, Ryan Hicks, Carol George, Andreas Ebneth, Oliver Keminer, Philip Gribbon, Barry Hardy, Stefanie Seltmann, Andreas Kurtz, Shawn Harmon, Jürgen Hescheler, Beate Kreisel, Hubrecht Institute for Developmental Biology and Stem Cell Research, and Publica
- Subjects
0301 basic medicine ,QH301-705.5 ,Process (engineering) ,media_common.quotation_subject ,Induced Pluripotent Stem Cells ,Biology ,Tissue procurement ,Cell Line ,03 medical and health sciences ,Resource (project management) ,Journal Article ,Humans ,Quality (business) ,Biology (General) ,Induced pluripotent stem cell ,lcsh:QH301-705.5 ,Biological Specimen Banks ,media_common ,Cryopreservation ,International research ,Medicine(all) ,Hot start ,General Medicine ,Cell Biology ,Europe ,Engineering management ,030104 developmental biology ,lcsh:Biology (General) ,Work flow ,Developmental Biology - Abstract
A fast track “Hot Start” process was implemented to launch the European Bank for Induced Pluripotent Stem Cells (EBiSC) to provide early release of a range of established control and disease linked human induced pluripotent stem cell (hiPSC) lines. Established practice amongst consortium members was surveyed to arrive at harmonised and publically accessible Standard Operations Procedures (SOPs) for tissue procurement, bio-sample tracking, iPSC expansion, cryopreservation, qualification and distribution to the research community. These were implemented to create a quality managed foundational collection of lines and associated data made available for distribution. Here we report on the successful outcome of this experience and work flow for banking and facilitating access to an otherwise disparate European resource, with lessons to benefit the international research community.eTOCThe report focuses on the EBiSC experience of rapidly establishing an operational capacity to procure, bank and distribute a foundational collection of established hiPSC lines. It validates the feasibility and defines the challenges of harnessing and integrating the capability and productivity of centres across Europe using commonly available resources currently in the field.
- Published
- 2017
40. Large mergers and acquisitions of European brewing groups-event study evidence on value creation
- Author
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Ebneth, Oliver and Theuvsen, Ludwig
- Subjects
Breweries -- Mergers, acquisitions and divestments ,Breweries -- Industry forecasts ,Breweries -- Market share ,Brewing industry -- Mergers, acquisitions and divestments ,Brewing industry -- Industry forecasts ,Brewing industry -- Market share ,Acquisitions and mergers -- Forecasts and trends ,Company acquisition/merger ,Company market share ,Market trend/market analysis ,Agricultural industry ,Business - Abstract
Event study analysis is used to examine 31 mergers and acquisitions among leading European brewing groups. It is seen that differences regarding the brewers' corporate success can be seen within the European peer group.
- Published
- 2007
41. MARK, a novel family of protein kinases that phosphorylate microtubule-associated proteins and trigger microtubule disruption
- Author
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Drewes, Gerard, Ebneth, Andreas, Preuss, Ute, Mandelkow, Eva-Maria, and Mandelkow, Eckhard
- Subjects
Protein kinases -- Research ,Phosphorylation -- Research ,Microtubules -- Research ,Biological sciences - Abstract
A study was conducted to describe the molecular cloning, distribution, activation mechanism and overexpression of two MARK proteins from rat. The findings indicate that MARKs phosphorylate microtubule-associated proteins and can potentially disrupt the microtubule network. Upstream factors may phosphorylate active MARKs which could provide a mechanism by which the stability of the cytoskeleton is controlled by extracellular signals.
- Published
- 1997
42. Generation of a set of isogenic, gene-edited iPSC lines homozygous for all main APOE variants and an APOE knock-out line
- Author
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Schmid, Benjamin, Prehn, Kennie R., Nimsanor, Natakarn, Garcia, Blanca Irene Aldana, Poulsen, Ulla, Jorring, Ida, Rasmussen, Mikkel A., Clausen, Christian, Mau-Holzmann, Ulrike A., Ramakrishna, Sarayu, Muddashetty, Ravi, Steeg, Rachel, Bruce, Kevin, Mackintosh, Peter, Ebneth, Andreas, Holst, Bjorn, Cabrera-Socorro, Alfredo, Schmid, Benjamin, Prehn, Kennie R., Nimsanor, Natakarn, Garcia, Blanca Irene Aldana, Poulsen, Ulla, Jorring, Ida, Rasmussen, Mikkel A., Clausen, Christian, Mau-Holzmann, Ulrike A., Ramakrishna, Sarayu, Muddashetty, Ravi, Steeg, Rachel, Bruce, Kevin, Mackintosh, Peter, Ebneth, Andreas, Holst, Bjorn, and Cabrera-Socorro, Alfredo
- Abstract
Alzheimer's disease (AD) is the most frequent neurodegenerative disease amongst the elderly. The SNPs rs429358 and rs7412 in the APOE gene are the most common risk factor for sporadic AD, and there are three different alleles commonly referred to as APOE-epsilon 2, APOE-epsilon 3 and APOE-epsilon 4. Induced pluripotent stem cells (iPSCs) hold great promise to model AD as such cells can be differentiated in vitro to the required cell type. Here we report the use of CRISPR/Cas9 technology employed on iPSCs from a healthy individual with an APOE-epsilon 3/epsilon 4 genotype to obtain isogenic APOE-epsilon 2/epsilon 2, APOE-epsilon 3/epsilon 3, APOE-epsilon 4/epsilon 4 lines as well as an APOE-knock-out line.
- Published
- 2019
43. SliceMap: an algorithm for automated brain region annotation
- Author
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Barbier, Michaël, Bottelbergs, A., Nuydens, R., Ebneth, A., De Vos, Winnok, Bottelbergs, Astrid, Nuydens, Rony, and Ebneth, Andreas
- Subjects
Statistics and Probability ,Tau pathology ,Matching (graph theory) ,Computer science ,Veterinary medicine ,Image processing ,Mice, Transgenic ,02 engineering and technology ,Biochemistry ,03 medical and health sciences ,Annotation ,Mice ,0302 clinical medicine ,Slice preparation ,Software ,0202 electrical engineering, electronic engineering, information engineering ,medicine ,Image Processing, Computer-Assisted ,Animals ,Molecular Biology ,Biology ,Computer. Automation ,business.industry ,Brain ,Computational Biology ,medicine.disease ,Computer Science Applications ,Computational Mathematics ,Brain region ,Chemistry ,Disease Models, Animal ,Computational Theory and Mathematics ,Tauopathies ,020201 artificial intelligence & image processing ,Tauopathy ,business ,Algorithm ,Engineering sciences. Technology ,030217 neurology & neurosurgery ,Mathematics ,Algorithms - Abstract
Summary Many neurodegenerative disorders, such as Alzheimer‘s Disease, pertain to or spread from specific sites of the brain. Hence, accurate disease staging or therapy assessment in transgenic model mice demands automated analysis of selected brain regions. To address this need, we have developed an algorithm, termed SliceMap, that enables contextual quantification by mapping anatomical information onto microtome-cut brain slices. For every newly acquired high-resolution image of a brain slice, the algorithm performs a coarse congealing-based registration to a library of pre-annotated reference slices. A subset of optimally matching reference slices is then used for refined, elastic registration. Morphotextural metrics are used to measure registration performance and to automatically detect poorly cut slices. We have implemented our method as a plugin for FIJI image analysis freeware, and we have used it to regionally quantify tau pathology in brain slices from a tauopathy (P301S) mouse model. By enabling region-based quantification, our method contributes to a more accurate assessment of neurodegenerative disease development. Availability and implementation The method is available as a plugin for FIJI from https://github.com/mbarbie1/SliceMap/, along with an example dataset and user instructions. Supplementary information Supplementary data are available at Bioinformatics online.
- Published
- 2017
44. Development of a Scalable, High-Throughput-Compatible Assay to Detect Tau Aggregates Using iPSC-Derived Cortical Neurons Maintained in a Three-Dimensional Culture Format
- Author
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Liesbeth Mertens, Alfredo Cabrera-Socorro, Alexis Bretteville, Ines Royaux, X. Medda, Sofie Versweyveld, L. Barnham, An Verheyen, Arjan Buist, Andreas Ebneth, and Annick Diels
- Subjects
0301 basic medicine ,Induced Pluripotent Stem Cells ,Cell ,Cell Culture Techniques ,Drug Evaluation, Preclinical ,tau Proteins ,Nanotechnology ,Protein aggregation ,Biology ,Biochemistry ,Analytical Chemistry ,Protein Aggregates ,03 medical and health sciences ,0302 clinical medicine ,High-Throughput Screening Assays ,medicine ,Humans ,Phosphorylation ,Cognitive decline ,Induced pluripotent stem cell ,Neurons ,Matrigel ,Neurodegeneration ,Brain ,medicine.disease ,030104 developmental biology ,medicine.anatomical_structure ,Tauopathies ,Cell culture ,Molecular Medicine ,Neuroscience ,030217 neurology & neurosurgery ,Biotechnology - Abstract
Tau aggregation is the pathological hallmark that best correlates with the progression of Alzheimer's disease (AD). The presence of neurofibrillary tangles (NFTs), formed of hyperphosphorylated tau, leads to neuronal dysfunction and loss, and is directly associated with the cognitive decline observed in AD patients. The limited success in targeting β-amyloid pathologies has reinforced the hypothesis of blocking tau phosphorylation, aggregation, and/or spreading as alternative therapeutic entry points to treat AD. Identification of novel therapies requires disease-relevant and scalable assays capable of reproducing key features of the pathology in an in vitro setting. Here we use induced pluripotent stem cells (iPSCs) as a virtually unlimited source of human cortical neurons to develop a robust and scalable tau aggregation model compatible with high-throughput screening (HTS). We downscaled cell culture conditions to 384-well plate format and used Matrigel to introduce an extra physical protection against cell detachment that reduces shearing stress and better recapitulates pathological conditions. We complemented the assay with AlphaLISA technology for the detection of tau aggregates in a high-throughput-compatible format. The assay is reproducible across users and works with different commercially available iPSC lines, representing a highly translational tool for the identification of novel treatments against tauopathies, including AD.
- Published
- 2016
45. Genetically Engineered iPSC-Derived FTDP-17 MAPT Neurons Display Mutation-Specific Neurodegenerative and Neurodevelopmental Phenotypes
- Author
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Verheyen, An, primary, Diels, Annick, additional, Reumers, Joke, additional, Van Hoorde, Kirsten, additional, Van den Wyngaert, Ilse, additional, van Outryve d’Ydewalle, Constantin, additional, De Bondt, An, additional, Kuijlaars, Jacobine, additional, De Muynck, Louis, additional, De Hoogt, Ronald, additional, Bretteville, Alexis, additional, Jaensch, Steffen, additional, Buist, Arjan, additional, Cabrera-Socorro, Alfredo, additional, Wray, Selina, additional, Ebneth, Andreas, additional, Roevens, Peter, additional, Royaux, Ines, additional, and Peeters, Pieter J., additional
- Published
- 2019
- Full Text
- View/download PDF
46. Generation of a set of isogenic, gene-edited iPSC lines homozygous for all main APOE variants and an APOE knock-out line
- Author
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Schmid, Benjamin, primary, Prehn, Kennie R., additional, Nimsanor, Natakarn, additional, Garcia, Blanca Irene Aldana, additional, Poulsen, Ulla, additional, Jørring, Ida, additional, Rasmussen, Mikkel A., additional, Clausen, Christian, additional, Mau-Holzmann, Ulrike A., additional, Ramakrishna, Sarayu, additional, Muddashetty, Ravi, additional, Steeg, Rachel, additional, Bruce, Kevin, additional, Mackintosh, Peter, additional, Ebneth, Andreas, additional, Holst, Bjørn, additional, and Cabrera-Socorro, Alfredo, additional
- Published
- 2019
- Full Text
- View/download PDF
47. Rapid establishment of the European Bank for induced Pluripotent Stem Cells (EBiSC) - the Hot Start experience
- Author
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De Sousa, Paul A., Steeg, Rachel, Wachter, Elisabeth, Bruce, Kevin, King, Jason, Hoeve, Marieke, Khadun, Shalinee, McConnachie, George, Holder, Julie, Kurtz, Andreas, Seltmann, Stefanie, Dewender, Johannes, Reimann, Sascha, Stacey, Glyn, O'Shea, Orla, Chapman, Charlotte, Healy, Lyn, Zimmermann, Heiko, Bolton, Bryan, Rawat, Trisha, Atkin, Isobel, Veiga, Anna, Kuebler, Bernd, Serano, Blanca Miranda, Saric, Tomo, Hescheler, Jürgen, Brüstle, Oliver, Peitz, Michael, Thiele, Cornelia, Geijsen, Niels, Holst, Bjørn, Clausen, Christian, Lako, Majlinda, Armstrong, Lyle, Gupta, Shailesh K., Kvist, Alexander J., Hicks, Ryan, Jonebring, Anna, Brolén, Gabriella, Ebneth, Andreas, Cabrera-Socorro, Alfredo, Foerch, Patrik, Geraerts, Martine, Stummann, Tina C., Harmon, Shawn, George, Carol, Streeter, Ian, Clarke, Laura, Parkinson, Helen, Harrison, Peter W., Faulconbridge, Adam, Cherubin, Luca, Burdett, Tony, Trigueros, Cesar, Patel, Minal J, Lucas, Christa, Hardy, Barry, Predan, Rok, Dokler, Joh, Brajnik, Maja, Keminer, Oliver, Pless, Ole, Gribbon, Philip, Claussen, Carsten, Ringwald, Annette, Kreisel, Beate, Courtney, Aidan, and Allsopp, Timothy E.
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- 2017
- Full Text
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48. Genetically Engineered iPSC-Derived FTDP-17 MAPT Neurons Display Mutation-Specific Neurodegenerative and Neurodevelopmental Phenotypes
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P. Roevens, Constantin d’Ydewalle, Andreas Ebneth, Jacobine Kuijlaars, Selina Wray, Annick Diels, Alexis Bretteville, Arjan Buist, An Verheyen, Joke Reumers, Alfredo Cabrera-Socorro, Ronald de Hoogt, Louis De Muynck, Ines Royaux, Kirsten Van Hoorde, An De Bondt, Ilse Van den Wyngaert, Pieter J. Peeters, and Steffen Jaensch
- Subjects
0301 basic medicine ,Tau protein ,medicine.disease_cause ,Biochemistry ,Article ,FTDP-17 ,03 medical and health sciences ,0302 clinical medicine ,mental disorders ,medicine ,Genetics ,MAPT ,Induced pluripotent stem cell ,ZFN ,lcsh:QH301-705.5 ,030304 developmental biology ,lcsh:R5-920 ,Mutation ,0303 health sciences ,iPSC ,biology ,Neurodegeneration ,Wnt signaling pathway ,Correction ,Cell Biology ,medicine.disease ,Zinc finger nuclease ,Phenotype ,Cell biology ,disease modelling ,030104 developmental biology ,lcsh:Biology (General) ,biology.protein ,lcsh:Medicine (General) ,030217 neurology & neurosurgery ,Frontotemporal dementia ,Developmental Biology - Abstract
Summary Tauopathies such as frontotemporal dementia (FTD) remain incurable to date, partially due to the lack of translational in vitro disease models. The MAPT gene, encoding the microtubule-associated protein tau, has been shown to play an important role in FTD pathogenesis. Therefore, we used zinc finger nucleases to introduce two MAPT mutations into healthy donor induced pluripotent stem cells (iPSCs). The IVS10+16 mutation increases the expression of 4R tau, while the P301S mutation is pro-aggregant. Whole-transcriptome analysis of MAPT IVS10+16 neurons reveals neuronal subtype differences, reduced neural progenitor proliferation potential, and aberrant WNT/SHH signaling. Notably, these neurodevelopmental phenotypes could be recapitulated in neurons from patients carrying the MAPT IVS10+16 mutation. Moreover, the additional pro-aggregant P301S mutation revealed additional phenotypes, such as an increased calcium burst frequency, reduced lysosomal acidity, tau oligomerization, and neurodegeneration. This series of iPSCs could serve as a platform to unravel a potential link between pathogenic 4R tau and FTD., Highlights • Analysis of ZFN-engineered MAPT IVS10+16 with or without additional P301S mutation • Neurodevelopmental phenotypes in ZFN and patient-derived MAPT IVS10+16 neurons • Neurodegenerative phenotypes in MAPT IVS10+16/P301S double-mutant neurons, In this article, Verheyen and colleagues introduced 2 MAPT mutations (IVS10+16/P301S) into healthy donor iPSCs. Characterization of MAPT IVS10+16 neurons revealed neurodevelopmental neuron subtype differences, reduced neural progenitor proliferation, and aberrant WNT/SHH signaling, while the additional pro-aggregant P301S mutation revealed phenotypes more related to neurodegeneration. These iPSCs could help to unravel a potential link between pathogenic 4R tau and FTD.
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- 2017
49. Generation of a set of isogenic, gene-edited iPSC lines homozygous for all main APOE variants and an APOE knock-out line
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Blanca Irene Aldana Garcia, Mikkel A. Rasmussen, Kennie R. Prehn, K. Bruce, Alfredo Cabrera-Socorro, Ulla B. Poulsen, Benjamin Schmid, Rachel Steeg, Peter Mackintosh, Bjørn Holst, Sarayu Ramakrishna, Ida Jørring, Ulrike A. Mau-Holzmann, Natakarn Nimsanor, Christian Clausen, Andreas Ebneth, and Ravi S. Muddashetty
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Male ,0301 basic medicine ,Apolipoprotein E ,Adolescent ,Induced Pluripotent Stem Cells ,Cell Culture Techniques ,Single-nucleotide polymorphism ,Biology ,Cell Line ,Gene Knockout Techniques ,03 medical and health sciences ,Apolipoproteins E ,0302 clinical medicine ,Genotype ,Humans ,CRISPR ,Allele ,Induced pluripotent stem cell ,lcsh:QH301-705.5 ,Gene ,Gene Editing ,Genetics ,Homozygote ,Cell Biology ,General Medicine ,3. Good health ,030104 developmental biology ,lcsh:Biology (General) ,Mutation ,lipids (amino acids, peptides, and proteins) ,030217 neurology & neurosurgery ,Developmental Biology - Abstract
Alzheimer's disease (AD) is the most frequent neurodegenerative disease amongst the elderly. The SNPs rs429358 and rs7412 in the APOE gene are the most common risk factor for sporadic AD, and there are three different alleles commonly referred to as APOE-epsilon 2, APOE-epsilon 3 and APOE-epsilon 4. Induced pluripotent stem cells (iPSCs) hold great promise to model AD as such cells can be differentiated in vitro to the required cell type. Here we report the use of CRISPR/Cas9 technology employed on iPSCs from a healthy individual with an APOE-epsilon 3/epsilon 4 genotype to obtain isogenic APOE-epsilon 2/epsilon 2, APOE-epsilon 3/epsilon 3, APOE-epsilon 4/epsilon 4 lines as well as an APOE-knock-out line.
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- 2019
50. Genetically Engineered iPSC-Derived FTDP-17 MAPT Neurons Display Mutation-Specific Neurodegenerative and Neurodevelopmental Phenotypes
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Verheyen, An, primary, Diels, Annick, additional, Reumers, Joke, additional, Van Hoorde, Kirsten, additional, Van den Wyngaert, Ilse, additional, van Outryve d’Ydewalle, Constantin, additional, De Bondt, An, additional, Kuijlaars, Jacobine, additional, De Muynck, Louis, additional, De Hoogt, Ronald, additional, Bretteville, Alexis, additional, Jaensch, Steffen, additional, Buist, Arjan, additional, Cabrera-Socorro, Alfredo, additional, Wray, Selina, additional, Ebneth, Andreas, additional, Roevens, Peter, additional, Royaux, Ines, additional, and Peeters, Pieter J., additional
- Published
- 2018
- Full Text
- View/download PDF
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