title: iPSC-derived neurons from GBA1-associated Parkinson's disease patients show autophagic defects and impaired calcium homeostasis.
abstract: Mutations in the acid β-glucocerebrosidase (GBA1) gene, responsible for the lysosomal storage disorder Gaucher's disease (GD), are the strongest genetic risk factor for Parkinson's disease (PD) known to date. Here we generate induced pluripotent stem cells from subjects with GD and PD harbouring GBA1 mutations, and differentiate them into midbrain dopaminergic neurons followed by enrichment using fluorescence-activated cell sorting. Neurons show a reduction in glucocerebrosidase activity and protein levels, increase in glucosylceramide and α-synuclein levels as well as autophagic and lysosomal defects. Quantitative proteomic profiling reveals an increase of the neuronal calcium-binding protein 2 (NECAB2) in diseased neurons. Mutant neurons show a dysregulation of calcium homeostasis and increased vulnerability to stress responses involving elevation of cytosolic calcium. Importantly, correction of the mutations rescues such pathological phenotypes. These findings provide evidence for a link between GBA1 mutations and complex changes in the autophagic/lysosomal system and intracellular calcium homeostasis, which underlie vulnerability to neurodegeneration.
authors.affiliation: 1] Department of Neurodegenerative Diseases, German Center for Neurodegenerative Diseases (DZNE), Tübingen 72076, Germany [2] Hertie-Institute for Clinical Brain Research, University of Tübingen, Tübingen 72076, Germany.
authors.forename: David C
authors.initials: DC
authors.lastname: Schöndorf
authors.affiliation: 1] Department of Medical Biotechnology and Translational Medicine, University of Milan, Milan 20090, Italy [2].
authors.forename: Massimo
authors.initials: M
authors.lastname: Aureli
authors.affiliation: 1] Department of Cell Biology, Harvard Medical School, Boston, Massacchusetts 02115, USA [2].
authors.forename: Fiona E
authors.initials: FE
authors.lastname: McAllister
authors.affiliation: 1] Emmy Noether-Group for Stem Cell Biology, Department of Molecular Embryology, Institute of Anatomy and Cell Biology, University of Freiburg, Freiburg 79104, Germany [2].
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Hindley
authors.affiliation: Werner Reichardt Center for Integrative Neuroscience (CIN), University of Tübingen, Tübingen 72076, Germany.
authors.forename: Florian
authors.initials: F
authors.lastname: Mayer
authors.affiliation: 1] Department of Neurodegenerative Diseases, German Center for Neurodegenerative Diseases (DZNE), Tübingen 72076, Germany [2] Hertie-Institute for Clinical Brain Research, University of Tübingen, Tübingen 72076, Germany.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Schmid
authors.affiliation: Genzyme, a Sanofi Company, Framingham, Massachusetts 01701, USA.
authors.forename: S Pablo
authors.initials: SP
authors.lastname: Sardi
authors.affiliation: Department of Medical Biotechnology and Translational Medicine, University of Milan, Milan 20090, Italy.
authors.forename: Manuela
authors.initials: M
authors.lastname: Valsecchi
authors.affiliation: 1] Department of Neurodegenerative Diseases, German Center for Neurodegenerative Diseases (DZNE), Tübingen 72076, Germany [2] Hertie-Institute for Clinical Brain Research, University of Tübingen, Tübingen 72076, Germany.
authors.forename: Susanna
authors.initials: S
authors.lastname: Hoffmann
authors.affiliation: 1] Department of Neurodegenerative Diseases, German Center for Neurodegenerative Diseases (DZNE), Tübingen 72076, Germany [2] Hertie-Institute for Clinical Brain Research, University of Tübingen, Tübingen 72076, Germany.
authors.forename: Lukas Kristoffer
authors.initials: LK
authors.lastname: Schwarz
authors.affiliation: 1] Hertie-Institute for Clinical Brain Research, University of Tübingen, Tübingen 72076, Germany [2] Department of Epileptology, Hertie Institute for Clinical Brain Research, University of Tübingen and German Center for Neurodegenerative Diseases, Tübingen 72076, Germany.
authors.forename: Ulrike
authors.initials: U
authors.lastname: Hedrich
authors.affiliation: 1] Department of Neurodegenerative Diseases, German Center for Neurodegenerative Diseases (DZNE), Tübingen 72076, Germany [2] Hertie-Institute for Clinical Brain Research, University of Tübingen, Tübingen 72076, Germany.
authors.forename: Daniela
authors.initials: D
authors.lastname: Berg
authors.affiliation: Genzyme, a Sanofi Company, Framingham, Massachusetts 01701, USA.
authors.forename: Lamya S
authors.initials: LS
authors.lastname: Shihabuddin
authors.affiliation: Werner Reichardt Center for Integrative Neuroscience (CIN), University of Tübingen, Tübingen 72076, Germany.
authors.forename: Jing
authors.initials: J
authors.lastname: Hu
authors.affiliation: 1] Emmy Noether-Group for Stem Cell Biology, Department of Molecular Embryology, Institute of Anatomy and Cell Biology, University of Freiburg, Freiburg 79104, Germany [2] Center for Biological Signaling Studies (BIOSS), University of Freiburg, Freiburg 79104 Germany.
authors.forename: Jan
authors.initials: J
authors.lastname: Pruszak
authors.affiliation: Department of Cell Biology, Harvard Medical School, Boston, Massacchusetts 02115, USA.
authors.forename: Steven P
authors.initials: SP
authors.lastname: Gygi
authors.affiliation: Department of Medical Biotechnology and Translational Medicine, University of Milan, Milan 20090, Italy.
authors.forename: Sandro
authors.initials: S
authors.lastname: Sonnino
authors.affiliation: 1] Department of Neurodegenerative Diseases, German Center for Neurodegenerative Diseases (DZNE), Tübingen 72076, Germany [2] Hertie-Institute for Clinical Brain Research, University of Tübingen, Tübingen 72076, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Gasser
authors.affiliation: 1] Department of Neurodegenerative Diseases, German Center for Neurodegenerative Diseases (DZNE), Tübingen 72076, Germany [2] Hertie-Institute for Clinical Brain Research, University of Tübingen, Tübingen 72076, Germany.
authors.forename: Michela
authors.initials: M
authors.lastname: Deleidi
mesh_terms: D001343:Autophagy; D002118:Calcium / Q000378:metabolism*; D002454:Cell Differentiation; D005962:Glucosylceramidase / Q000235:genetics*; D006026:Glycoside Hydrolases / Q000378:metabolism; D006706:Homeostasis; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000201:enzymology / Q000473:pathology*; D009474:Neurons / Q000201:enzymology / Q000473:pathology*; D010300:Parkinson Disease / Q000276:immunology / Q000378:metabolism / Q000473:pathology*
chemical_list: D006026:Glycoside Hydrolases; D005962:Glucosylceramidase; D002118:Calcium