Pathogenic Pathways in Early-Onset Autosomal Recessive Parkinson's Disease Discovered Using Isogenic Human Dopaminergic Neurons

Autor: J. Wade Harper, Alban Ordureau, Tobias Grass, Silvia Piccinotti, Lee L. Rubin, Lily Sarrafha, Fumiki Yanagawa, Takayuki Uozumi, Yasujiro Kiyota, Christina Bell, Joao A. Paulo, Chicheng Sun, Tim Ahfeldt, Gustavo M. Parfitt
Rok vydání: 2020
Předmět:
Proteomics
0301 basic medicine
Parkinson's disease
Proteome
medicine.disease_cause
Biochemistry
Parkin
Transcriptome
transcriptomics
0302 clinical medicine
disease modeling
Gene Knock-In Techniques
human pluripotent stem cells
ATP13A2
Induced pluripotent stem cell
lcsh:QH301-705.5
Genetics
lcsh:R5-920
Dopaminergic
Parkinson Disease
Mitochondria
Phenotype
CRISPR
lcsh:Medicine (General)
Signal Transduction
Pluripotent Stem Cells
Tyrosine 3-Monooxygenase
Genes
Recessive
Biology
Article
Cell Line
03 medical and health sciences
medicine
Humans
genome editing
Genetic Predisposition to Disease
Gene
Genetic Association Studies
Dopaminergic Neurons
DJ1
PARK7
Cell Biology
medicine.disease
nervous system diseases
030104 developmental biology
lcsh:Biology (General)
Mutation
030217 neurology & neurosurgery
Oxidative stress
Developmental Biology
Zdroj: Stem Cell Reports, Vol 14, Iss 1, Pp 75-90 (2020)
Stem Cell Reports
ISSN: 2213-6711
DOI: 10.1016/j.stemcr.2019.12.005
Popis: Summary Parkinson's disease (PD) is a complex and highly variable neurodegenerative disease. Familial PD is caused by mutations in several genes with diverse and mostly unknown functions. It is unclear how dysregulation of these genes results in the relatively selective death of nigral dopaminergic neurons (DNs). To address this question, we modeled PD by knocking out the PD genes PARKIN (PRKN), DJ-1 (PARK7), and ATP13A2 (PARK9) in independent isogenic human pluripotent stem cell (hPSC) lines. We found increased levels of oxidative stress in all PD lines. Increased death of DNs upon differentiation was found only in the PARKIN knockout line. Using quantitative proteomics, we observed dysregulation of mitochondrial and lysosomal function in all of the lines, as well as common and distinct molecular defects caused by the different PD genes. Our results suggest that precise delineation of PD subtypes will require evaluation of molecular and clinical data.
Highlights • CRISPR knockin of reporter in TH locus allows live tracking and isolation of DNs • Large-scale 3D midbrain DN differentiation using spinner flask culture • Phenotypic comparison of isogenic DNs harboring knockouts of PARKIN, DJ-1, or ATP13A2 • Transcriptomics and quantitative proteomics studies determine common and distinct PD pathways
In this article, Ahfeldt, Rubin, and colleagues model Parkinson's disease (PD) in human pluripotent stem cells by knocking out PARKIN, DJ-1, or ATP13A2. They report increased levels of oxidative stress in all PD lines and death of dopaminergic neurons in the PARKIN-KO. Using transcriptomics and quantitative proteomics approaches they determine common and distinct molecular defects caused by different PD genes.
Databáze: OpenAIRE
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