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| Status |
Public on Mar 01, 2018 |
| Title |
Parkinson’s Disease Genetic Risk in a Midbrain Neuronal Cell Line |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
Genome binding/occupancy profiling by high throughput sequencing
|
| Summary |
In genome-wide association studies of complex diseases, many risk polymorphisms are found to lie in non-coding DNA and likely confer risk through allele-dependent differences in gene regulatory elements. However, because distal regulatory elements can alter gene expression at various distances on linear DNA, the identity of relevant genes is unknown for most risk loci. In Parkinson’s disease, at least some genetic risk is likely intrinsic to a neuronal subpopulation of cells in the brain regions affected. In order to compare neuron-relevant methods of pairing risk polymorphisms to target genes as well as to further characterize a single-cell model of a neurodegenerative disease, we used the portionally-dopaminergic, neuronal, mesencephalic-derived cell line LUHMES to dissect differentiation-specific mechanisms of gene expression. We compared genome-wide gene expression in undifferentiated and differentiated cells with genome-wide histone H3K27ac and CTCF-bound regions. Whereas promoters and CTCF binding were largely consistent between differentiated and undifferentiated cells, enhancers were mostly unique. We matched the differentiation-specific appearance or disappearance of enhancers with changes in gene expression and identified 22,057 enhancers paired with 6,388 differentially expressed genes by proximity. These enhancers are enriched with at least 13 transcription factor response elements, driving a cluster of genes involved in neurogenesis. We show that differentiated LUHMES cells, but not undifferentiated cells, show enrichment for PD-risk SNPs. Candidate genes for these loci are largely unrelated, though a subset is linked to synaptic vesicle cycling and transport, implying that PD-related disruption of these pathways is intrinsic to dopaminergic neurons.
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| Overall design |
Lund human mesencephalic cells (LUHMES) can be propagated as cycling undifferentiated neural progenitor cells or induced to differentiate into dopaminergic neurons. We cultured and propagated LUHMES cells (undifferentiated cells) and induced differentiation for 6 days (differentiated cells). We isolated RNA from undifferentiated and differentiated cells in order to compare gene expression (RNA-seq), CTCF binding location (ChIP-seq), and enhancer/promoter (H3K27Ac ChIP-seq) activity in the two states.
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| Contributor(s) |
Coetzee GA, Pierce SE, Tyson T |
| Citation(s) |
29486295 |
| |
| Submission date |
Jan 26, 2018 |
| Last update date |
Mar 26, 2019 |
| Contact name |
Gerry A. Coetzee |
| E-mail(s) |
gerry.coetzee@vai.org
|
| Phone |
6162345305
|
| Organization name |
Van Andel Research Institute
|
| Department |
Center for Neurodegenerative Science
|
| Lab |
Gerry Coetzee
|
| Street address |
333 Bostwick Ave NE
|
| City |
Grand Rapids |
| State/province |
MI |
| ZIP/Postal code |
49503 |
| Country |
USA |
| |
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| Platforms (1) |
| GPL18573 |
Illumina NextSeq 500 (Homo sapiens) |
|
| Samples (20)
|
| GSM2948717 |
LUHMES undifferentiated RNA-seq Sample 1 |
| GSM2948718 |
LUHMES undifferentiated RNA-seq Sample 2 |
| GSM2948719 |
LUHMES undifferentiated RNA-seq Sample 3 |
| GSM2948720 |
LUHMES undifferentiated RNA-seq Sample 4 |
| GSM2948721 |
LUHMES undifferentiated RNA-seq Sample 5 |
| GSM2948722 |
LUHMES undifferentiated RNA-seq Sample 6 |
| GSM2948723 |
LUHMES differentiated RNA-seq Sample 1 |
| GSM2948724 |
LUHMES differentiated RNA-seq Sample 2 |
| GSM2948725 |
LUHMES differentiated RNA-seq Sample 3 |
| GSM2948726 |
LUHMES differentiated RNA-seq Sample 4 |
| GSM2948727 |
LUHMES differentiated RNA-seq Sample 5 |
| GSM2948728 |
LUHMES differentiated RNA-seq Sample 6 |
| GSM2948729 |
LUHMES undifferentiated ChIP-seq H3k27ac Sample 1 |
| GSM2948730 |
LUHMES undifferentiated ChIP-seq H3k27ac Sample 2 |
| GSM2948731 |
LUHMES differentiated ChIP-seq H3k27ac Sample 1 |
| GSM2948732 |
LUHMES differentiated ChIP-seq H3k27ac Sample 2 |
| GSM2948733 |
LUHMES undifferentiated ChIP-seq CTCF Sample 1 |
| GSM2948734 |
LUHMES undifferentiated ChIP-seq CTCF Sample 2 |
| GSM2948735 |
LUHMES differentiated ChIP-seq CTCF Sample 1 |
| GSM2948736 |
LUHMES differentiated ChIP-seq CTCF Sample 2 |
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| Relations |
| BioProject |
PRJNA431771 |
| SRA |
SRP131485 |
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