title: A New Glucocerebrosidase Chaperone Reduces α-Synuclein and Glycolipid Levels in iPSC-Derived Dopaminergic Neurons from Patients with Gaucher Disease and Parkinsonism.
abstract: Among the known genetic risk factors for Parkinson disease, mutations in GBA1, the gene responsible for the lysosomal disorder Gaucher disease, are the most common. This genetic link has directed attention to the role of the lysosome in the pathogenesis of parkinsonism. To study how glucocerebrosidase impacts parkinsonism and to evaluate new therapeutics, we generated induced human pluripotent stem cells from four patients with Type 1 (non-neuronopathic) Gaucher disease, two with and two without parkinsonism, and one patient with Type 2 (acute neuronopathic) Gaucher disease, and differentiated them into macrophages and dopaminergic neurons. These cells exhibited decreased glucocerebrosidase activity and stored the glycolipid substrates glucosylceramide and glucosylsphingosine, demonstrating their similarity to patients with Gaucher disease. Dopaminergic neurons from patients with Type 2 and Type 1 Gaucher disease with parkinsonism had reduced dopamine storage and dopamine transporter reuptake. Levels of α-synuclein, a protein present as aggregates in Parkinson disease and related synucleinopathies, were selectively elevated in neurons from the patients with parkinsonism or Type 2 Gaucher disease. The cells were then treated with NCGC607, a small-molecule noninhibitory chaperone of glucocerebrosidase identified by high-throughput screening and medicinal chemistry structure optimization. This compound successfully chaperoned the mutant enzyme, restored glucocerebrosidase activity and protein levels, and reduced glycolipid storage in both iPSC-derived macrophages and dopaminergic neurons, indicating its potential for treating neuronopathic Gaucher disease. In addition, NCGC607 reduced α-synuclein levels in dopaminergic neurons from the patients with parkinsonism, suggesting that noninhibitory small-molecule chaperones of glucocerebrosidase may prove useful for the treatment of Parkinson disease.
  
  
  
  Because GBA1 mutations are the most common genetic risk factor for Parkinson disease, dopaminergic neurons were generated from iPSC lines derived from patients with Gaucher disease with and without parkinsonism. These cells exhibit deficient enzymatic activity, reduced lysosomal glucocerebrosidase levels, and storage of glucosylceramide and glucosylsphingosine. Lines generated from the patients with parkinsonism demonstrated elevated levels of α-synuclein. To reverse the observed phenotype, the neurons were treated with a novel noninhibitory glucocerebrosidase chaperone, which successfully restored glucocerebrosidase activity and protein levels and reduced glycolipid storage. In addition, the small-molecule chaperone reduced α-synuclein levels in dopaminergic neurons, indicating that chaperoning glucocerebrosidase to the lysosome may provide a novel therapeutic strategy for both Parkinson disease and neuronopathic forms of Gaucher disease.
authors.affiliation: Section of Molecular Neurogenetics, National Human Genome Research Institute.
authors.forename: Elma
authors.initials: E
authors.lastname: Aflaki
authors.affiliation: Section of Molecular Neurogenetics, National Human Genome Research Institute.
authors.forename: Daniel K
authors.initials: DK
authors.lastname: Borger
authors.affiliation: Section of Molecular Neurogenetics, National Human Genome Research Institute.
authors.forename: Nima
authors.initials: N
authors.lastname: Moaven
authors.affiliation: Section of Molecular Neurogenetics, National Human Genome Research Institute.
authors.forename: Barbara K
authors.identifier: 0000-0002-5044-7017
authors.initials: BK
authors.lastname: Stubblefield
authors.affiliation: University of Kansas Specialized Chemistry Center, University of Kansas, Lawrence, Kansas 66047, and.
authors.forename: Steven A
authors.initials: SA
authors.lastname: Rogers
authors.affiliation: National Center for Advancing Translational Sciences.
authors.forename: Samarjit
authors.initials: S
authors.lastname: Patnaik
authors.affiliation: University of Kansas Specialized Chemistry Center, University of Kansas, Lawrence, Kansas 66047, and.
authors.forename: Frank J
authors.initials: FJ
authors.lastname: Schoenen
authors.affiliation: Section of Molecular Neurogenetics, National Human Genome Research Institute.
authors.forename: Wendy
authors.initials: W
authors.lastname: Westbroek
authors.affiliation: National Center for Advancing Translational Sciences.
authors.forename: Wei
authors.identifier: 0000-0003-1034-0757
authors.initials: W
authors.lastname: Zheng
authors.affiliation: Clinical Neurocardiology Section, and.
authors.forename: Patricia
authors.initials: P
authors.lastname: Sullivan
authors.affiliation: Diabetic Cardiovascular Disease Center and Department of Medicine, Washington University School of Medicine, St. Louis, Missouri 63110.
authors.forename: Hideji
authors.initials: H
authors.lastname: Fujiwara
authors.affiliation: Diabetic Cardiovascular Disease Center and Department of Medicine, Washington University School of Medicine, St. Louis, Missouri 63110.
authors.forename: Rohini
authors.initials: R
authors.lastname: Sidhu
authors.affiliation: Cellular Neurophysiology Unit, National Institute of Neurological Disease and Stroke, National Institutes of Health, Bethesda, Maryland 20892.
authors.forename: Zayd M
authors.identifier: 0000-0002-1445-1457
authors.initials: ZM
authors.lastname: Khaliq
authors.affiliation: Section of Molecular Neurogenetics, National Human Genome Research Institute.
authors.forename: Grisel J
authors.identifier: 0000-0001-7785-2197
authors.initials: GJ
authors.lastname: Lopez
authors.affiliation: Clinical Neurocardiology Section, and.
authors.forename: David S
authors.identifier: 0000-0002-5709-9940
authors.initials: DS
authors.lastname: Goldstein
authors.affiliation: Diabetic Cardiovascular Disease Center and Department of Medicine, Washington University School of Medicine, St. Louis, Missouri 63110.
authors.forename: Daniel S
authors.initials: DS
authors.lastname: Ory
authors.affiliation: National Center for Advancing Translational Sciences.
authors.forename: Juan
authors.initials: J
authors.lastname: Marugan
authors.affiliation: Section of Molecular Neurogenetics, National Human Genome Research Institute, sidranse@mail.nih.gov.
authors.forename: Ellen
authors.identifier: 0000-0002-3019-8500
authors.initials: E
authors.lastname: Sidransky
mesh_terms: D000083:Acetanilides / Q000494:pharmacology; D001549:Benzamides / Q000494:pharmacology; D002395:Catecholamines / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics; D059290:Dopaminergic Neurons / Q000187:drug effects / Q000378:metabolism*; D005260:Female; D005776:Gaucher Disease / Q000473:pathology*; D005962:Glucosylceramidase; D005963:Glucosylceramides / Q000037:antagonists & inhibitors* / Q000378:metabolism; D006017:Glycolipids / Q000378:metabolism*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000187:drug effects* / Q000378:metabolism; D053444:Inhibitory Postsynaptic Potentials / Q000187:drug effects / Q000235:genetics; D052119:Lysosomal-Associated Membrane Protein 2 / Q000378:metabolism; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects / Q000235:genetics; D009154:Mutation / Q000235:genetics; D020734:Parkinsonian Disorders / Q000473:pathology*; D018408:Patch-Clamp Techniques; D051844:alpha-Synuclein / Q000378:metabolism*; D001617:beta-Glucosidase / Q000235:genetics
chemical_list: D000083:Acetanilides; D001549:Benzamides; D002395:Catecholamines; D005963:Glucosylceramides; D006017:Glycolipids; D052119:Lysosomal-Associated Membrane Protein 2; C000621316:NCGC607; D051844:alpha-Synuclein; D001617:beta-Glucosidase; C517537:GBA2 protein, human; D005962:Glucosylceramidase
keywords: dopaminergic neurons; glucocerebrosidase; induced pluripotent stem cells; parkinsonism; pharmacological chaperone; α-synuclein