Lysosomal Dysfunction in iPS Cell-derived Dopaminergic Neurons Generated from a Patient with Monogenic Parkinson's Disease (PARK9) — The Association Specialists
  1. Schedule
  2. Small Things with Big Impact: Nanoparticles, Exosomes and MicroRNA
  3. Lysosomal Dysfunction in iPS Cell-derived Dopaminergic Neurons Generated from a Patient with Monogenic Parkinson's Disease (PARK9)

Lysosomal Dysfunction in iPS Cell-derived Dopaminergic Neurons Generated from a Patient with Monogenic Parkinson's Disease (PARK9) (309)

Nicholas F Blair 1 , Joseph R Mazzulli 2 , Taiji Tsunemi 2 , Chistopher W Vaughan 1 , Ernst J Wolvetang 3 , Carolyn M Sue 1 , Dimitri Krainc 2
  1. Kolling Institute of Medical Research, St Leonards, NSW, Australia
  2. MassGeneral Institute for Neurodegeneration, Department of Neurology, Massachusetts General Hospital, Harvard Medical School , Boston, MA, United States
  3. Australian Institute of Bioengineering & Nanotechnology, University of Queensland, Brisbane, Qld, Australia

Background

The discovery of monogenic forms of Parkinson's Disease has lead to important insights into the pathogenesis of the disease. One of these monogenic conditions is Kufor-Rakeb Syndrome (KRS), denoted by the locus PARK9, which causes autosomal recessive juvenile-onset Parkinsonism. It has recently been found to be associated with mutations in ATP13A2, a gene which encodes a putative lysosomal P-type ATPase, the specific function of which remains unknown. There is initial evidence to suggest ATP13A2 may be involved in the processing and degradation of α-synuclein, a protein that has a central role in the development of Parkinson's disease.

Objective

Our aim was to generate a human dopaminergic cell model derived from a patient with KRS for use in studying the pathophysiology of the condition, and to further elucidate the role of lysosomal dysfunction and its relationship to α-synuclein accumulation in Parkinson's Disease.

Methods

Mutations in ATP13A2 were identified using direct sequencing of the gene in a 22 year old male with a clinical presentation consistent with KRS. Induced pluripotent stem (iPS) cells were generated from a skin biopsy contributed by the patient using published methods. The iPS cells were then differentiated into dopaminergic neurons using recently described protocols. The cells were analysed using immunocytochemistry, live cell imaging and Western blotting.

Results

We successfully generated neural cells that expressed the characteristic markers of midbrain dopaminergic neurons, FoxA2 and LmX1a. We found evidence of lysosomal dysfunction and significant α-Synuclein accumulation in ATP13A2-mutant neurons.

Conclusion

This study confirmed the feasibility of deriving dopaminergic neurons from iPS cells as a cellular model of a monogenic form of PD. The results demonstrate the importance of ATP13A2 in lysosomal function and support the hypothesis that ATP13A2 plays a role in the degradation of α-Synuclein.