Objective: The aim of this project is to investigate post-mortem brain tissues, iPSC and iPSC-derived neurons from controls and patients with DYT1 dystonia to find transcriptomic signatures that underlie the abnormal pathways in patients.

Background: Dystonia is a disabling movement disorder characterised by involuntary muscle contractions causing abnormal posturing and twisting of one or more parts of the body. The cause of dystonia is not known and is best described as a circuitopathy without neurodegeneration. The commonest genetic form is caused by mutations in TOR1A gene (DYT1) with early-onset severe generalised dystonia.

Method: A total of 10 cell lines and 6 brain donors were used in this project. Fibroblasts from 5 controls and 5 symptomatic DYT1 patients were reprogramed using episomal plasmid transduction. The iPSC generated were differentiated to cortical and medium spiny neurons. Once the cells reached maturity (100 DIV and 80 DIV, respectively), RNA was extracted from all cell types and brain tissues (frontal cortex and basal ganglia) using TRI Reagent® solution. All samples were assessed with RNA integrity and quality controls and sent to Novogene Co for the transcriptome profiling, single nucleotide polymorphism identification, and differential gene expression analysis.

Results: The results showed a distributed sample correlation between all cell types and brain samples, indicating that the differentiated cells were representative. The differential expression between controls and patients showed a high number of up and down regulated genes in all neurons and brain tissues. The annotation and the enrichment between all cell lines and brain tissues suggested the potential of 96 genes downregulated and 73 upregulated, highlighting the pathways involved.

Conclusion: Our preliminary data identified differences in gene expression between DYT1 patients and healthy controls in different neuronal cell types and brain tissue samples. Further analyses are ongoing to refine the numbers of genes and hone in on those that show the greatest change and have biological relevance for dystonia and will be presented. The study of the candidate genes that highlight potential cellular pathways will be followed by functional analysis helping to elucidate cell specific pathways involved in dystonia for further therapeutic targets and design of biomarkers in future clinical trials.

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MDS Abstracts - https://www.mdsabstracts.org/abstract/transcriptomic-profiling-in-dyt1-dystonia-unravelling-pathogenic-pathways/