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| Status |
Public on Aug 17, 2022 |
| Title |
Downstream Effects of Mutations in SOD1 and TARDBP Converge on Gene Expression Impairment in Patient-Derived Motor Neurons |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
Non-coding RNA profiling by high throughput sequencing
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| Summary |
We established iPSCs from healthy donors, SOD1-ALS and TDP43-ALS patients. Using our differentiation protocol originally developed by Reinhardt et al.,2013, we diferentiated these iPSCs toward spinal motor neurons (MNs) and reproduce ALS pathology in a dish. To extend our understanding of finding different molecular mechanisms and pathways related to SOD1- and TDP43 mutations in ALS disease, we have performed a comprehensive gene expression profiling study using RNA-Seq of the iPSC-derived MN models from control individuals and carefully compared with those from SOD1-ALS and TDP43-ALS patients.
To generate novel hypotheses of putative underlying molecular mechanisms in ALS, we used human induced pluripotent stem cell (hiPSCs)-derived motor neurons (MNs) from SOD1- and TARDBP (TDP-43 protein)-mutant-ALS patients and healthy controls to perform high-throughput RNA-sequencing (RNA-Seq). An integrated bioinformatics approach was employed to identify differentially expressed genes (DEGs) and key pathways underlying these familial forms of the disease (fALS). In TDP43-ALS, we found dysregulation of transcripts encoding components of the transcriptional machinery and transcripts involved in splicing regulation were particularly affected. In contrast, less is known about the role of SOD1 in RNA metabolism in motor neurons. Here we found that many transcripts relevant for mitochondrial function were specifically altered in SOD1-ALS, indicating transcriptional signatures and expression patterns can vary significantly depending on the causal gene that is mutated. Surprisingly, however, we identified a clear downregulation of genes involved in protein translation in SOD1-ALS suggesting that ALS-causing SOD1 mutations shift cellular RNA abundance profiles to cause neural dysfunction. Altogether, we provided here an extensive profiling of mRNA expression in two ALS models at the cellular level, corroborating the major role of RNA metabolism and protein translation as a common pathomechanism in ALS
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| Overall design |
We isolated total RNA from iPSC-derived MNs (healthy donor: 3 samples, SOD1-ALS: 2 samples, TDP43-ALS: 3 samples. Total 8 samples) for the gene expression RNA-Seqanalysis (Illumina HiSeq 2500). All of the iPSC lines we used were low passage number (less than 20) and analysis were performed at DIV 30. We isolated total RNA from iPSC-derived MNs (healthy donor: 3 samples, SOD1-ALS: 2 samples, TDP43-ALS: 3 samples. Total 8 samples) and performed small RNA-seq (Illumina HiSeq 2500). All of the iPSC lines we used were low passage number (less than 20) and analysis were performed at DIV 30-45.
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| Contributor(s) |
Hermann A, Freischmidt A, Dash BP |
| Citation(s) |
36077049, 38676626 |
| |
| Submission date |
Aug 10, 2022 |
| Last update date |
May 31, 2024 |
| Contact name |
Andreas Hermann |
| E-mail(s) |
Andreas.Hermann@med.uni-rostock.de
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| Organization name |
University Medical Center Rostock
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| Department |
Neurology
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| Lab |
Neurodegeneration
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| Street address |
Gehlsheimer Str.20
|
| City |
Rostock |
| State/province |
Mecklenburg-Western Pomerania |
| ZIP/Postal code |
18147 |
| Country |
Germany |
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| Platforms (1) |
| GPL16791 |
Illumina HiSeq 2500 (Homo sapiens) |
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| Samples (16)
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| Relations |
| BioProject |
PRJNA868391 |
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