Induced Accelerated Aging in Induced Pluripotent Stem Cell Lines from Patients with Parkinson's Disease.

摘要

Background: iPS reprogramming to model 'disease-in-a-dish has become an attractive approach to study disease mechanisms. However, a frank pathophysiological phenotype in iPSC-derived PD neurons remains to be shown. Objective/Hypothesis: The objective of this proposal is to accelerate the aging process of iPS-derived dopaminergic neurons with the goal of reproducing a Parkinson s disease (PD) specific phenotype in vitro. Specific Aim 1. To generate vector constructs and to establish Dox-inducible iPSC lines that express the Hutchinson-Gilford progeria (HGPS) gene (mutant lamin A with an in- frame loss of 150 nucleotides) and differentiate them into dopaminergic neurons. Deliverable of this aim is the introduction of mutant lamin A in iPSC lines and optimization of inducible expression of mutant lamin A in iPSCs and during the differentiation into dopaminergic neurons. Specific Aim 2. To test whether iPSC-derived dopaminergic neurons transduced with mutant lamin A show changes in age-regulated genes at the mRNA and protein levels at different time- points during differentiation, thus exhibiting an accelerated aging. Deliverable of this aim is the assessment of specific age-related aging pattern of gene and protein expression in induced mutant lamin A modified iPSC-derived neurons. Specific Aim 3. To test whether induced mutant lamin A cell lines differentiated into dopaminergic neurons exhibit hallmark pathology of PD, such as protein aggregation of alpha-synuclein, posttranslational modification, and signs of mitochondrial pathology. Deliverable of this aim is the assessment of the pathological PD-related phenotype in induced mutant lamin A modified iPSC- derived neurons.