Quantitative Proteomic Analysis of Primitive Neural Stem Cells from LRRK2 G2019S-Associated Parkinson’s Disease Patient-Derived iPSCs

Abstract

Parkinson’s disease (PD) is a common neurodegenerative disease, causing movement defects. The incidence of PD is constantly increasing and this disease is still incurable. Thus, understanding PD pathophysiology would be pivotal for the development of PD therapy, and various PD models have thus been already developed. Through recent advances in reprogramming techniques, a primitive neural stem cell (pNSC) derived from PD patient induced pluripotent stem cells (iPSCs) could be potentially used as a reproducible and reliable experimental system to analyze the effect of the leucine-rich repeat kinase 2 G2019S mutation (LK2GS) in neural cells. Here, we investigated the advantages of such a model system through quantitative proteomic analysis of pNSCs from normal control iPSCs and familial PD patient iPSCs harboring LK2GS. We confirmed that the expression of molecules known to be involved in PD pathogenesis, such as oxidative stress-, cell adhesion-, and cytoskeleton-related proteins, were altered in the LK2GS pNSC. In addition, we showed that down-regulation of Ku80, which was found in the proteomic analysis with LK2GS pNSCs, resulted in apoptosis induced by DNA damage response. Taken together, we suggest that pNSCs from PD iPSCs could provide a reliable and useful model system to study PD. Moreover, the highly expandable pNSC is suitable for multi-omics approaches to understand PD pathologies and discover therapeutic targets for PD.

Transcriptome analysis in LRRK2 and idiopathic Parkinson’s disease at different glucose levels

“…The identified expression changes associated with PD in idiopathic and monogenic L2PD involved alterations related to the plasma membrane, extracellular matrix, cell adhesion, and neural processes. These results are in line with previous studies reporting deregulation in the cytoskeleton, cell adhesion, or extracellular matrix processes in various PD patients biospecimens such as blood cells 9 , 10 , iPSC-derived DAn 11 , 12 , or post-mortem substantia nigra 13 . Specifically, the previous transcriptomic study available in PD fibroblasts was done in patients carrying PRKN mutations 14 .…”

Induced pluripotent stem cell-based organ-on-a-chip as personalized drug screening tools: A focus on neurodegenerative disorders

“…In the case of PD, dopaminergic (DA) neurons, that are the most affected cell type, were obtained from the differentiation of iPSCs generated from healthy subjects, 65,66 non-familial PD 67 and familial PD patients carrying various mutation. [68][69][70][71][72][73] In iPSC-derived DA neurons, the capacity of cysteamine to decrease α-synuclein levels and reduce 6-OHDA toxicity, previously observed in mice, was confirmed. 68 These cells were also informative on mechanisms associated with PD mitochondrial disfunction and oxidative stress, showing that coenzyme Q10, rapamycin, and LRRK2 kinase inhibitor GW5074 rescued valinomycin or concanamycin A induced vulnerability.…”

Ciliogenesis is Not Directly Regulated by LRRK2 Kinase Activity in Neurons

“…Established human iPSCs were fed TeSR TM -E8 TM (STEMCELL Technologies, Vancouver, BC, Canada) according to the manufacturer’s recommendations. Neural stem cells (NSCs) were differentiated from human iPSCs as previously described [ 22 ] with some modifications [ 23 ]. Human iPSCs were seeded on GeltrexTM LDEV-free Reduced Growth Factor Basement Membrane Matrix-coated dishes at about 20% confluency.…”