title: A new glucocerebrosidase-deficient neuronal cell model provides a tool to probe pathophysiology and therapeutics for Gaucher disease.
abstract: Glucocerebrosidase is a lysosomal hydrolase involved in the breakdown of glucosylceramide. Gaucher disease, a recessive lysosomal storage disorder, is caused by mutations in the gene GBA1 Dysfunctional glucocerebrosidase leads to accumulation of glucosylceramide and glycosylsphingosine in various cell types and organs. Mutations in GBA1 are also a common genetic risk factor for Parkinson disease and related synucleinopathies. In recent years, research on the pathophysiology of Gaucher disease, the molecular link between Gaucher and Parkinson disease, and novel therapeutics, have accelerated the need for relevant cell models with GBA1 mutations. Although induced pluripotent stem cells, primary rodent neurons, and transfected neuroblastoma cell lines have been used to study the effect of glucocerebrosidase deficiency on neuronal function, these models have limitations because of challenges in culturing and propagating the cells, low yield, and the introduction of exogenous mutant GBA1 To address some of these difficulties, we established a high yield, easy-to-culture mouse neuronal cell model with nearly complete glucocerebrosidase deficiency representative of Gaucher disease. We successfully immortalized cortical neurons from embryonic null allele gba(-/-) mice and the control littermate (gba(+/+)) by infecting differentiated primary cortical neurons in culture with an EF1α-SV40T lentivirus. Immortalized gba(-/-) neurons lack glucocerebrosidase protein and enzyme activity, and exhibit a dramatic increase in glucosylceramide and glucosylsphingosine accumulation, enlarged lysosomes, and an impaired ATP-dependent calcium-influx response; these phenotypical characteristics were absent in gba(+/+) neurons. This null allele gba(-/-) mouse neuronal model provides a much-needed tool to study the pathophysiology of Gaucher disease and to evaluate new therapies.
authors.affiliation: Section on Molecular Neurogenetics, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Wendy
authors.initials: W
authors.lastname: Westbroek
authors.affiliation: Section on Molecular Neurogenetics, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Matthew
authors.initials: M
authors.lastname: Nguyen
authors.affiliation: Section on Molecular Neurogenetics, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA Postgraduate Program in Cellular and Molecular Biology, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS 91501-970, Brazil.
authors.forename: Marina
authors.initials: M
authors.lastname: Siebert
authors.affiliation: Section on Molecular Neurogenetics, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Taylor
authors.initials: T
authors.lastname: Lindstrom
authors.affiliation: Section on Molecular Neurogenetics, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Robert A
authors.initials: RA
authors.lastname: Burnett
authors.affiliation: Section on Molecular Neurogenetics, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Elma
authors.initials: E
authors.lastname: Aflaki
authors.affiliation: Section on Molecular Neurogenetics, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Olive
authors.initials: O
authors.lastname: Jung
authors.affiliation: Section on Molecular Neurogenetics, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Rafael
authors.initials: R
authors.lastname: Tamargo
authors.affiliation: Genomics, Development, and Disease Section, Genetic Disease Research Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Jorge L
authors.initials: JL
authors.lastname: Rodriguez-Gil
authors.affiliation: Biostrategies LC, State University, AR 72467, USA.
authors.forename: Walter
authors.initials: W
authors.lastname: Acosta
authors.affiliation: Laboratory of Experimental Cancer Research, Department of Radiation Oncology and Experimental Cancer Research, Ghent University Hospital, Ghent 9000, Belgium.
authors.forename: An
authors.initials: A
authors.lastname: Hendrix
authors.affiliation: Section on Molecular Neurogenetics, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Bahafta
authors.initials: B
authors.lastname: Behre
authors.affiliation: Section on Molecular Neurogenetics, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Nahid
authors.initials: N
authors.lastname: Tayebi
authors.affiliation: Washington University School of Medicine, St. Louis, MO 63110-1093, USA.
authors.forename: Hideji
authors.initials: H
authors.lastname: Fujiwara
authors.affiliation: Washington University School of Medicine, St. Louis, MO 63110-1093, USA.
authors.forename: Rohini
authors.initials: R
authors.lastname: Sidhu
authors.affiliation: Cell Biology Section, Neurogenetics Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Benoit
authors.initials: B
authors.lastname: Renvoise
authors.affiliation: Lysosomal Disorders Treatment and Research Program, Clinical Labs, University of Massachusetts Medical School, Worcester, MA 01655, USA.
authors.forename: Edward I
authors.initials: EI
authors.lastname: Ginns
authors.affiliation: Cytogenetics Core, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Amalia
authors.initials: A
authors.lastname: Dutra
authors.affiliation: Cytogenetics Core, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Evgenia
authors.initials: E
authors.lastname: Pak
authors.affiliation: Biostrategies LC, State University, AR 72467, USA.
authors.forename: Carole
authors.initials: C
authors.lastname: Cramer
authors.affiliation: Washington University School of Medicine, St. Louis, MO 63110-1093, USA.
authors.forename: Daniel S
authors.initials: DS
authors.lastname: Ory
authors.affiliation: Genomics, Development, and Disease Section, Genetic Disease Research Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: William J
authors.initials: WJ
authors.lastname: Pavan
authors.affiliation: Section on Molecular Neurogenetics, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA sidranse@mail.nih.gov.
authors.forename: Ellen
authors.identifier: 0000-0002-3019-8500
authors.initials: E
authors.lastname: Sidransky
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000378:metabolism; D051927:CD24 Antigen / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002461:Cell Line, Transformed; D002478:Cells, Cultured; D005776:Gaucher Disease / Q000201:enzymology / Q000503:physiopathology* / Q000628:therapy*; D005962:Glucosylceramidase / Q000172:deficiency* / Q000378:metabolism; D007621:Karyotyping; D008247:Lysosomes / Q000378:metabolism; D008810:Mice, Inbred C57BL; D008954:Models, Biological; D009474:Neurons / Q000201:enzymology* / Q000473:pathology*; D020648:Peptide Elongation Factor 1 / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D013379:Substrate Specificity
chemical_list: D000952:Antigens, Polyomavirus Transforming; D051927:CD24 Antigen; D020648:Peptide Elongation Factor 1; D000255:Adenosine Triphosphate; D005962:Glucosylceramidase; D002118:Calcium
keywords: Gaucher disease; Glucocerebrosidase; Glucosylceramide; Glucosylsphingosine; Neuron