Neuronopathic Gaucher's disease: induced pluripotent stem cells for disease modelling and testing chaperone activity of small compounds

Hum Mol Genet. 2013 Feb 15;22(4):633-45. doi: 10.1093/hmg/dds471. Epub 2012 Oct 31.

Abstract

Gaucher's disease (GD) is caused by mutations in the GBA1 gene, which encodes acid-β-glucosidase, an enzyme involved in the degradation of complex sphingolipids. While the non-neuronopathic aspects of the disease can be treated with enzyme replacement therapy (ERT), the early-onset neuronopathic form currently lacks therapeutic options and is lethal. We have developed an induced pluripotent stem cell (iPSc) model of neuronopathic GD. Dermal fibroblasts of a patient with a P.[LEU444PRO];[GLY202ARG] genotype were transfected with a loxP-flanked polycistronic reprogramming cassette consisting of Oct4, Sox2, Klf4 and c-Myc and iPSc lines derived. A non-integrative lentiviral vector expressing Cre recombinase was used to eliminate the reprogramming cassette from the reprogrammed cells. Our GD iPSc express pluripotent markers, differentiate into the three germ layers, form teratomas, have a normal karyotype and show the same mutations and low acid-β-glucosidase activity as the original fibroblasts they were derived from. We have differentiated them efficiently into neurons and also into macrophages without observing deleterious effects of the mutations on the differentiation process. Using our system as a platform to test chemical compounds capable of increasing acid-β-glucosidase activity, we confirm that two nojirimycin analogues can rescue protein levels and enzyme activity in the cells affected by the disease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 1-Deoxynojirimycin / analogs & derivatives*
  • 1-Deoxynojirimycin / pharmacology
  • Adamantane / analogs & derivatives*
  • Adamantane / pharmacology
  • Antigens, Differentiation / metabolism
  • Base Sequence
  • Cell Differentiation
  • Cells, Cultured
  • DNA Mutational Analysis
  • Dopaminergic Neurons / enzymology
  • Drug Evaluation, Preclinical
  • Enzyme Stability / drug effects
  • Gaucher Disease / drug therapy*
  • Gaucher Disease / pathology
  • Gene Expression
  • Glucosylceramidase / genetics
  • Glucosylceramidase / metabolism
  • Humans
  • Induced Pluripotent Stem Cells / drug effects*
  • Induced Pluripotent Stem Cells / enzymology
  • Induced Pluripotent Stem Cells / physiology
  • Kruppel-Like Factor 4
  • Lysosomes / enzymology
  • Macrophages / metabolism
  • Oligonucleotide Array Sequence Analysis
  • Protein Transport
  • Small Molecule Libraries
  • Transcriptome

Substances

  • 5-N,6-O-(N''-octyliminomethylidene)nojirimycin
  • 5-N,6-S-(N''-4'-(adamantane-1-carbonylamino)butyliminomethylidene)-6-thionojirimycin
  • Antigens, Differentiation
  • KLF4 protein, human
  • Kruppel-Like Factor 4
  • Small Molecule Libraries
  • 1-Deoxynojirimycin
  • Glucosylceramidase
  • Adamantane