Juan Antonio García-León1, Alfredo Cabrera-Socorro2, Kristel Eggermont3, Ann Swijsen4, Joke Terryn5, Raheem Fazal6, FatemehArefeh Nami3, Laura Ordovás3, Ana Quiles3, Frederic Lluis3, Lutgarde Serneels7, Keimpe Wierda4, Annerieke Sierksma7, Mohamed Kreir2, Francisco Pestana2, Philip Van Damme8, Bart De Strooper9, Lieven Thorrez10, Andreas Ebneth2, Catherine M Verfaillie11. 1. Department of Development and Regeneration, Stem Cell Biology and Embryology, KU Leuven Stem Cell Institute, Leuven, Belgium. Electronic address: jgarleon@hotmail.com. 2. Janssen Research & Development, a Division of Janssen Pharmaceutica N.V., Beerse, Belgium. 3. Department of Development and Regeneration, Stem Cell Biology and Embryology, KU Leuven Stem Cell Institute, Leuven, Belgium. 4. KU Leuven-Department of Neurosciences, Experimental Neurology and Leuven Research Institute for Neuroscience and Disease (LIND), Leuven, Belgium; VIB Center for Brain and Disease Research, Laboratory of Neurobiology, Leuven, Belgium. 5. Department of Development and Regeneration, Stem Cell Biology and Embryology, KU Leuven Stem Cell Institute, Leuven, Belgium; VIB Center for Brain and Disease Research, Laboratory of Neurobiology, Leuven, Belgium. 6. Department of Development and Regeneration, Stem Cell Biology and Embryology, KU Leuven Stem Cell Institute, Leuven, Belgium; KU Leuven-Department of Neurosciences, Experimental Neurology and Leuven Research Institute for Neuroscience and Disease (LIND), Leuven, Belgium; VIB Center for Brain and Disease Research, Laboratory of Neurobiology, Leuven, Belgium. 7. VIB Center for the Biology of Disease, Leuven, Belgium Center for Human Genetics, Universitaire ziekenhuizen and LIND, KU, Leuven, Belgium. 8. KU Leuven-Department of Neurosciences, Experimental Neurology and Leuven Research Institute for Neuroscience and Disease (LIND), Leuven, Belgium; VIB Center for Brain and Disease Research, Laboratory of Neurobiology, Leuven, Belgium; University Hospitals Leuven, Department of Neurology, Leuven, Belgium. 9. VIB Center for the Biology of Disease, Leuven, Belgium Center for Human Genetics, Universitaire ziekenhuizen and LIND, KU, Leuven, Belgium; Institute of Neurology, University College London, London, UK. 10. Tissue Engineering Laboratory, Department of Development and Regeneration, KU Leuven, Campus Kulak Kortrijk, Kortrijk, Belgium. 11. Department of Development and Regeneration, Stem Cell Biology and Embryology, KU Leuven Stem Cell Institute, Leuven, Belgium; KU Leuven-Department of Neurosciences, Experimental Neurology and Leuven Research Institute for Neuroscience and Disease (LIND), Leuven, Belgium. Electronic address: catherine.verfaillie@kuleuven.be.
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.
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.
Authors: Justine D Manos; Christina N Preiss; Nandini Venkat; Joseph Tamm; Peter Reinhardt; Taekyung Kwon; Jessica Wu; Allison D Winter; Thomas R Jahn; Kiran Yanamandra; Katherine Titterton; Eric Karran; Xavier Langlois Journal: iScience Date: 2021-12-18
Authors: Juan Antonio Garcia-Leon; Laura Caceres-Palomo; Elisabeth Sanchez-Mejias; Marina Mejias-Ortega; Cristina Nuñez-Diaz; Juan Jose Fernandez-Valenzuela; Raquel Sanchez-Varo; Jose Carlos Davila; Javier Vitorica; Antonia Gutierrez Journal: Int J Mol Sci Date: 2020-09-18 Impact factor: 5.923