A new glucocerebrosidase-deficient neuronal cell model provides a tool to probe pathophysiology and therapeutics for Gaucher disease

Wendy Westbroek; Matthew Nguyen; Marina Siebert; Taylor Lindstrom; Robert A. Burnett; Elma Aflaki; Olive Jung; Rafael Tamargo; Jorge L. Rodriguez-Gil; Walter Acosta; An Hendrix; Bahafta Behre; Nahid Tayebi; Hideji Fujiwara; Rohini Sidhu; Benoit Renvoise; Edward I. Ginns; Amalia Dutra; Evgenia Pak; Carole Cramer; Daniel S. Ory; William J. Pavan; Ellen Sidransky

doi:10.1242/dmm.024588

Disease Models & Mechanisms (Jul 2016)

A new glucocerebrosidase-deficient neuronal cell model provides a tool to probe pathophysiology and therapeutics for Gaucher disease

Wendy Westbroek,
Matthew Nguyen,
Marina Siebert,
Taylor Lindstrom,
Robert A. Burnett,
Elma Aflaki,
Olive Jung,
Rafael Tamargo,
Jorge L. Rodriguez-Gil,
Walter Acosta,
An Hendrix,
Bahafta Behre,
Nahid Tayebi,
Hideji Fujiwara,
Rohini Sidhu,
Benoit Renvoise,
Edward I. Ginns,
Amalia Dutra,
Evgenia Pak,
Carole Cramer,
Daniel S. Ory,
William J. Pavan,
Ellen Sidransky

Affiliations

Wendy Westbroek: Section on Molecular Neurogenetics, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA
Matthew Nguyen: Section on Molecular Neurogenetics, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA
Marina Siebert: Section on Molecular Neurogenetics, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA
Taylor Lindstrom: Section on Molecular Neurogenetics, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA
Robert A. Burnett: Section on Molecular Neurogenetics, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA
Elma Aflaki: Section on Molecular Neurogenetics, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA
Olive Jung: Section on Molecular Neurogenetics, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA
Rafael Tamargo: Section on Molecular Neurogenetics, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA
Jorge L. Rodriguez-Gil: Genomics, Development, and Disease Section, Genetic Disease Research Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA
Walter Acosta: Biostrategies LC, State University, AR 72467, USA
An Hendrix: Laboratory of Experimental Cancer Research, Department of Radiation Oncology and Experimental Cancer Research, Ghent University Hospital, Ghent 9000, Belgium
Bahafta Behre: Section on Molecular Neurogenetics, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA
Nahid Tayebi: Section on Molecular Neurogenetics, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA
Hideji Fujiwara: Washington University School of Medicine, St. Louis, MO 63110-1093, USA
Rohini Sidhu: Washington University School of Medicine, St. Louis, MO 63110-1093, USA
Benoit Renvoise: Cell Biology Section, Neurogenetics Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD 20892, USA
Edward I. Ginns: Lysosomal Disorders Treatment and Research Program, Clinical Labs, University of Massachusetts Medical School, Worcester, MA 01655, USA
Amalia Dutra: Cytogenetics Core, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA
Evgenia Pak: Cytogenetics Core, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA
Carole Cramer: Biostrategies LC, State University, AR 72467, USA
Daniel S. Ory: Washington University School of Medicine, St. Louis, MO 63110-1093, USA
William J. Pavan: Genomics, Development, and Disease Section, Genetic Disease Research Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA
Ellen Sidransky: Section on Molecular Neurogenetics, Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, Bethesda, MD 20892, USA

DOI: https://doi.org/10.1242/dmm.024588
Journal volume & issue: Vol. 9, no. 7
pp. 769 – 778

Abstract

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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.

Published in Disease Models & Mechanisms

ISSN: 1754-8403 (Print); 1754-8411 (Online)
Publisher: The Company of Biologists
Country of publisher: United Kingdom
LCC subjects: Medicine: Pathology
Website: https://journals.biologists.com/dmm

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