Poster Presentation 6th Annual Meeting for Australasian Society for Stem Cell Research 2013

Using human induced neural precursor cells to model Parkinson's disease. (#162)

Rebecca Playne 1 , Erin Firmin 1 , Kathryn Jones 1 , Bronwen Connor 1
  1. University of Auckland, Auckland, New Zealand

Parkinson’s disease (PD) is a progressive neurodegenerative disorder affecting 1-2% of people over the age of 65. PD pathology is characterised by the degeneration of nigrostriatal dopaminergic neurons and the deposition of intracellular proteinacious inclusions called Lewy bodies, resulting in motor and non-motor symptoms. At present there are no therapies that slow or halt disease progression and current treatments only target disease symptoms. The study of PD has been hampered by a lack of access to live affected human neurons. Recently induced pluripotent stem cell (iPSC) technology has been used to develop human cellular models for PD. These models are limited by a low efficiency of iPS generation (<0.01% by retroviral transduction) and low yields of authentic midbrain dopaminergic neurons following differentiation. Direct reprogramming techniques offer an efficient alternative strategy for modelling PD using patient-specific cells. Several groups have directly reprogrammed rodent and human foetal fibroblasts into induced neural precursor (iNP) cells, however we are the only group to have directly generated iNPs from adult human fibroblasts (efficiency: 0.05% by plasmid transfection). We have demonstrated that transient expression of the neural genes SOX2 and PAX6 by plasmid transfection using Lipofectamine LTX generates iNPs from fibroblasts from a patient with a common PD-causing LRRK2 missense mutation. These iNPs endogenously expressed neural stem/progenitor markers OCT4, SOX2, PAX6, BMI1, FOXG1 and pro-neural genes NCAM1, NGN2. Additionally, preliminary evidence suggests iNPs can be efficiently differentiated into neurons co-expressing the catecholaminergic marker TH and neuronal markers TUJ1 and NSE (>50% of Tuj1+ neurons co-expressed TH). These findings support the development of a PD cell model that could be used to investigate early cellular and molecular alterations contributing to PD pathogenesis and to screen for disease-modifying agents