Determination of mRNA expression profile for Flp-In T-REx 293 cells inducibly expressing ΔE302/3 Torsin1a for 72 hours

Dystonia is a centrally driven movement disorder characterized by often painful twisting motions and postures. The majority of dystonia cases are idiopathic and sporadic, although a significant number of rare inherited forms have defined genetic causes. However, regardless of etiology, the biological mechanisms for nearly all forms of dystonia are still largely unknown and there are currently no disease-modifying treatments. Here we show a common role for impaired functioning of the eIF2α stress-response pathway in rare inherited and common sporadic forms of dystonia. We first identified a potential role for the eIF2α pathway in dystonia through a genome-wide RNAi screen aimed at correcting a DYT1 dystonia-related cellular phenotype. Mechanistically, we find that the corrective effect is mediated by activating the eIF2α pathway through an eIF2α-phosphorylation and ATF4 transcription factor-dependent mechanism. We further find evidence that this pathway is impaired in DYT1 patient-derived cell lines, suggesting a role in pathogenesis. Notably, pathogenicity of reduced eIF2α pathway signaling is supported by another known cause of dystonia, DYT16, which is due to loss-of-function mutations in the PRKRA gene that encodes an eIF2α kinase activator. Here, we demonstrate that sporadic cervical dystonia patients share a rare, conserved, loss-of-function ATF4 mutation. Finally, we show that pharmacologically enhancing eIF2α signaling improves DYT1-related phenotypes in both a cultured cell system and a mouse model. Our results identify eIF2α signaling as a novel pathway in the pathogenesis of multiple forms of dystonia. Our work also shows that the eIF2α pathway constitutes an attractive biological target for the development of dystonia therapeutics. 3 independent biological samples were plated and induced with tetracyline (5µg/mL) for 72hr prior to cell harvest and total RNA extraction.