title: ER Stress and Autophagic Perturbations Lead to Elevated Extracellular α-Synuclein in GBA-N370S Parkinson's iPSC-Derived Dopamine Neurons.
abstract: Heterozygous mutations in the glucocerebrosidase gene (GBA) represent the strongest common genetic risk factor for Parkinson's disease (PD), the second most common neurodegenerative disorder. However, the molecular mechanisms underlying this association are still poorly understood. Here, we have analyzed ten independent induced pluripotent stem cell (iPSC) lines from three controls and three unrelated PD patients heterozygous for the GBA-N370S mutation, and identified relevant disease mechanisms. After differentiation into dopaminergic neurons, we observed misprocessing of mutant glucocerebrosidase protein in the ER, associated with activation of ER stress and abnormal cellular lipid profiles. Furthermore, we observed autophagic perturbations and an enlargement of the lysosomal compartment specifically in dopamine neurons. Finally, we found increased extracellular α-synuclein in patient-derived neuronal culture medium, which was not associated with exosomes. Overall, ER stress, autophagic/lysosomal perturbations, and elevated extracellular α-synuclein likely represent critical early cellular phenotypes of PD, which might offer multiple therapeutic targets.
authors.affiliation: Oxford Parkinson's Disease Centre, University of Oxford, South Parks Road, Oxford OX1 3QX, UK; Department of Physiology, Anatomy and Genetics, University of Oxford, South Parks Road, Oxford OX1 3QX, UK.
authors.forename: Hugo J R
authors.initials: HJ
authors.lastname: Fernandes
authors.affiliation: Oxford Parkinson's Disease Centre, University of Oxford, South Parks Road, Oxford OX1 3QX, UK; Department of Physiology, Anatomy and Genetics, University of Oxford, South Parks Road, Oxford OX1 3QX, UK.
authors.forename: Elizabeth M
authors.initials: EM
authors.lastname: Hartfield
authors.affiliation: Department of Physiology, Anatomy and Genetics, University of Oxford, South Parks Road, Oxford OX1 3QX, UK.
authors.forename: Helen C
authors.initials: HC
authors.lastname: Christian
authors.affiliation: Division of Basic Neurosciences, Biomedical Research Foundation of the Academy of Athens, Athens 11526, Greece.
authors.forename: Evangelia
authors.initials: E
authors.lastname: Emmanoulidou
authors.affiliation: Oxford Parkinson's Disease Centre, University of Oxford, South Parks Road, Oxford OX1 3QX, UK; Nuffield Department of Clinical Medicine, Division of Clinical Neurology, University of Oxford, Oxford OX3 9DU, UK.
authors.forename: Ying
authors.initials: Y
authors.lastname: Zheng
authors.affiliation: Oxford Parkinson's Disease Centre, University of Oxford, South Parks Road, Oxford OX1 3QX, UK; Department of Physiology, Anatomy and Genetics, University of Oxford, South Parks Road, Oxford OX1 3QX, UK.
authors.forename: Heather
authors.initials: H
authors.lastname: Booth
authors.affiliation: Oxford Parkinson's Disease Centre, University of Oxford, South Parks Road, Oxford OX1 3QX, UK; Department of Physiology, Anatomy and Genetics, University of Oxford, South Parks Road, Oxford OX1 3QX, UK; Institute of Molecular Medicine, University of Southern Denmark, Odense 5230, Denmark.
authors.forename: Helle
authors.initials: H
authors.lastname: Bogetofte
authors.affiliation: Oxford Parkinson's Disease Centre, University of Oxford, South Parks Road, Oxford OX1 3QX, UK; Department of Physiology, Anatomy and Genetics, University of Oxford, South Parks Road, Oxford OX1 3QX, UK.
authors.forename: Charmaine
authors.initials: C
authors.lastname: Lang
authors.affiliation: Oxford Parkinson's Disease Centre, University of Oxford, South Parks Road, Oxford OX1 3QX, UK; Department of Physiology, Anatomy and Genetics, University of Oxford, South Parks Road, Oxford OX1 3QX, UK.
authors.forename: Brent J
authors.initials: BJ
authors.lastname: Ryan
authors.affiliation: Genzyme, a Sanofi Company, Framingham, MA 01701, USA.
authors.forename: S Pablo
authors.initials: SP
authors.lastname: Sardi
authors.affiliation: Oxford Parkinson's Disease Centre, University of Oxford, South Parks Road, Oxford OX1 3QX, UK; Department of Physiology, Anatomy and Genetics, University of Oxford, South Parks Road, Oxford OX1 3QX, UK.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Badger
authors.affiliation: Oxford Parkinson's Disease Centre, University of Oxford, South Parks Road, Oxford OX1 3QX, UK; The James Martin Stem Cell Facility, Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford OX1 3RE, UK.
authors.forename: Jane
authors.initials: J
authors.lastname: Vowles
authors.affiliation: Oxford Parkinson's Disease Centre, University of Oxford, South Parks Road, Oxford OX1 3QX, UK; Nuffield Department of Clinical Medicine, Division of Clinical Neurology, University of Oxford, Oxford OX3 9DU, UK.
authors.forename: Samuel
authors.initials: S
authors.lastname: Evetts
authors.affiliation: Oxford Parkinson's Disease Centre, University of Oxford, South Parks Road, Oxford OX1 3QX, UK; Nuffield Department of Clinical Medicine, Division of Clinical Neurology, University of Oxford, Oxford OX3 9DU, UK.
authors.forename: George K
authors.initials: GK
authors.lastname: Tofaris
authors.affiliation: Division of Basic Neurosciences, Biomedical Research Foundation of the Academy of Athens, Athens 11526, Greece.
authors.forename: Kostas
authors.initials: K
authors.lastname: Vekrellis
authors.affiliation: Oxford Parkinson's Disease Centre, University of Oxford, South Parks Road, Oxford OX1 3QX, UK; Nuffield Department of Clinical Medicine, Division of Clinical Neurology, University of Oxford, Oxford OX3 9DU, UK.
authors.forename: Kevin
authors.initials: K
authors.lastname: Talbot
authors.affiliation: Oxford Parkinson's Disease Centre, University of Oxford, South Parks Road, Oxford OX1 3QX, UK; Nuffield Department of Clinical Medicine, Division of Clinical Neurology, University of Oxford, Oxford OX3 9DU, UK.
authors.forename: Michele T
authors.initials: MT
authors.lastname: Hu
authors.affiliation: Oxford Parkinson's Disease Centre, University of Oxford, South Parks Road, Oxford OX1 3QX, UK; The James Martin Stem Cell Facility, Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford OX1 3RE, UK.
authors.forename: William
authors.initials: W
authors.lastname: James
authors.affiliation: Oxford Parkinson's Disease Centre, University of Oxford, South Parks Road, Oxford OX1 3QX, UK; The James Martin Stem Cell Facility, Sir William Dunn School of Pathology, University of Oxford, South Parks Road, Oxford OX1 3RE, UK.
authors.forename: Sally A
authors.initials: SA
authors.lastname: Cowley
authors.affiliation: Oxford Parkinson's Disease Centre, University of Oxford, South Parks Road, Oxford OX1 3QX, UK; Department of Physiology, Anatomy and Genetics, University of Oxford, South Parks Road, Oxford OX1 3QX, UK. Electronic address: richard.wade-martins@dpag.ox.ac.uk.
authors.forename: Richard
authors.initials: R
authors.lastname: Wade-Martins
mesh_terms: D000818:Animals; D001343:Autophagy; D002460:Cell Line; D002478:Cells, Cultured; D059290:Dopaminergic Neurons / Q000166:cytology / Q000378:metabolism*; D059865:Endoplasmic Reticulum Stress; D055354:Exosomes / Q000378:metabolism; D005962:Glucosylceramidase / Q000235:genetics*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D008247:Lysosomes / Q000378:metabolism; D051379:Mice; D020125:Mutation, Missense; D055495:Neurogenesis; D010300:Parkinson Disease / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D051844:alpha-Synuclein / Q000378:metabolism*
chemical_list: D051844:alpha-Synuclein; D005962:Glucosylceramidase