title: Impaired APP activity and altered Tau splicing in embryonic stem cell-derived astrocytes obtained from an APPsw transgenic minipig.
abstract: Animal models of familial juvenile onset of Alzheimer's disease (AD) often fail to produce diverse pathological features of the disease by modification of single gene mutations that are responsible for the disease. They can hence be poor models for testing and development of novel drugs. Here, we analyze in vitro-produced stem cells and their derivatives from a large mammalian model of the disease created by overexpression of a single mutant human gene (APPsw). We produced hemizygous and homozygous radial glial-like cells following culture and differentiation of embryonic stem cells (ESCs) isolated from embryos obtained from mated hemizygous minipigs. These cells were confirmed to co-express varying neural markers, including NES, GFAP and BLBP, typical of type one radial glial cells (RGs) from the subgranular zone. These cells had altered expression of CCND1 and NOTCH1 and decreased expression of several ribosomal RNA genes. We found that these cells were able to differentiate into astrocytes upon directed differentiation. The astrocytes produced had decreased α- and β-secretase activity, increased γ-secretase activity and altered splicing of tau. This indicates novel aspects of early onset mechanisms related to cell renewal and function in familial AD astrocytes. These outcomes also highlight that radial glia could be a potentially useful population of cells for drug discovery, and that altered APP expression and altered tau phosphorylation can be detected in an in vitro model of the disease. Finally, it might be possible to use large mammal models to model familial AD by insertion of only a single mutation.
authors.affiliation: Department of Veterinary Clinical and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, DK-1870 Frederiksberg, Denmark vh@sund.ku.dk.
authors.forename: Vanessa J
authors.initials: VJ
authors.lastname: Hall
authors.affiliation: Department of Veterinary Disease Biology, Faculty of Health and Medical Sciences, University of Copenhagen, DK-1870 Frederiksberg, Denmark.
authors.forename: Maiken M
authors.initials: MM
authors.lastname: Lindblad
authors.affiliation: Department of Biomedicine, Aarhus University, Faculty of Health, DK-8000 Aarhus, Denmark.
authors.forename: Jannik E
authors.initials: JE
authors.lastname: Jakobsen
authors.affiliation: Department of Biomedicine, Aarhus University, Faculty of Health, DK-8000 Aarhus, Denmark.
authors.forename: Anders
authors.initials: A
authors.lastname: Gunnarsson
authors.affiliation: Department of Large Animal Sciences, Faculty of Life Sciences, University of Copenhagen, DK-1870 Frederiksberg, Denmark.
authors.forename: Mette
authors.initials: M
authors.lastname: Schmidt
authors.affiliation: Bioneer Biomedical Technology, DK-2970 Hørsholm, Denmark.
authors.forename: Mikkel A
authors.initials: MA
authors.lastname: Rasmussen
authors.affiliation: Center for Biotechnology and Biomedicine, Institute of Bioanalytical Chemistry, University of Leipzig, 04103 Leipzig, Germany.
authors.forename: Daniela
authors.initials: D
authors.lastname: Volke
authors.affiliation: Octapharma Biopharmaceuticals GmbH, 69120 Heidelberg, Germany.
authors.forename: Thole
authors.initials: T
authors.lastname: Zuchner
authors.affiliation: Department of Veterinary Clinical and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, DK-1870 Frederiksberg, Denmark.
authors.forename: Poul
authors.initials: P
authors.lastname: Hyttel
mesh_terms: D053829:Amyloid Precursor Protein Secretases / Q000378:metabolism; D016229:Amyloid beta-Peptides / Q000378:metabolism*; D000818:Animals; D030801:Animals, Genetically Modified; D001253:Astrocytes / Q000166:cytology*; D002454:Cell Differentiation; D053595:Embryonic Stem Cells / Q000378:metabolism*; D006801:Humans; D009154:Mutation / Q000235:genetics*; D055495:Neurogenesis; D009457:Neuroglia / Q000378:metabolism; D010766:Phosphorylation; D012326:RNA Splicing / Q000235:genetics*; D012335:RNA, Ribosomal / Q000096:biosynthesis; D015398:Signal Transduction; D013552:Swine; D013556:Swine, Miniature; D016875:tau Proteins / Q000235:genetics*
chemical_list: D016229:Amyloid beta-Peptides; D012335:RNA, Ribosomal; D016875:tau Proteins; D053829:Amyloid Precursor Protein Secretases
keywords: APP Swedish mutation; Alzheimer's disease; Astrocytes; Astrogenesis; Cell cycle; Pig; Radial glial cells; Secretase expression; Tau phosphorylation