Intermediate filament protein accumulation in motor neurons derived from giant axonal neuropathy iPSCs rescued by restoration of gigaxonin

Hum Mol Genet. 2015 Mar 1;24(5):1420-31. doi: 10.1093/hmg/ddu556. Epub 2014 Nov 4.

Abstract

Giant axonal neuropathy (GAN) is a progressive neurodegenerative disease caused by autosomal recessive mutations in the GAN gene resulting in a loss of a ubiquitously expressed protein, gigaxonin. Gene replacement therapy is a promising strategy for treatment of the disease; however, the effectiveness and safety of gigaxonin reintroduction have not been tested in human GAN nerve cells. Here we report the derivation of induced pluripotent stem cells (iPSCs) from three GAN patients with different GAN mutations. Motor neurons differentiated from GAN iPSCs exhibit accumulation of neurofilament (NF-L) and peripherin (PRPH) protein and formation of PRPH aggregates, the key pathological phenotypes observed in patients. Introduction of gigaxonin either using a lentiviral vector or as a stable transgene resulted in normalization of NEFL and PRPH levels in GAN neurons and disappearance of PRPH aggregates. Importantly, overexpression of gigaxonin had no adverse effect on survival of GAN neurons, supporting the feasibility of gene replacement therapy. Our findings demonstrate that GAN iPSCs provide a novel model for studying human GAN neuropathologies and for the development and testing of new therapies in relevant cell types.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Axons
  • Cell Differentiation
  • Cells, Cultured
  • Cytoskeletal Proteins / genetics
  • Cytoskeletal Proteins / metabolism*
  • Fibroblasts / cytology
  • Fibroblasts / metabolism
  • Genetic Therapy / methods
  • Genetic Vectors / genetics
  • Giant Axonal Neuropathy / genetics*
  • Giant Axonal Neuropathy / therapy
  • Humans
  • Induced Pluripotent Stem Cells / cytology*
  • Intermediate Filament Proteins / genetics*
  • Intermediate Filaments / genetics
  • Intermediate Filaments / metabolism
  • Karyotyping
  • Lentivirus / genetics
  • Motor Neurons / cytology
  • Motor Neurons / metabolism*
  • Mutation
  • Phenotype

Substances

  • Cytoskeletal Proteins
  • GAN protein, human
  • Intermediate Filament Proteins