Dr Emmanouil Metzakopian
The Wellcome Trust Sanger Institute
United Kingdom
CRISPR-Cas9 genetic screens for genes conferring survival to oxidative stress in dopamine neurons
Parkinson's UK
294,293
01/11/2015
3
There are currently no disease halting treatments for Parkinson’s disease (PD). With the recent failure of numerous neuroprotective therapies for PD at the clinical testing stage (Phytopharm’s Cogane, Ceregene’s AAV-Neurturin, etc), there is a critical need for identifying novel neuroprotective pathways. Large scale whole-genome, random mutation analysis (using the CRISPR-Cas9 system) represents one of the most efficient methods by which unidentified molecular mechanisms can be discovered. Working in Prof Allen Bradley’s lab at the Wellcome Trust Sanger Institute, I have utilised this approach to identify interesting new targets in specific cancers – highlighting novel protective functions in specific genes. I would now like to apply this technology to midbrain dopamine (DA) neurons with the goal of determining novel neuroprotective genes. By stressing cultured DA neurons with hydrogen-peroxide and rotenone, and then sequencing DNA extracted from the surviving cells, I hope to identify the mutations that infer either vulnerability or strength in the cells. Upon validating all of the newly identified targets, I will generate mutant mice for behavioural analysis and PD modelling in collaboration with Prof Roger Barker’s lab at the University of Cambridge. In parallel with the behavioural testing, I will replicate the oxidative stress screens in human ES cell-derived DA neuron, allowing me to identify conserved neuroprotective mechanisms. The goal of this work will be to identify a range novel pathways which can be further investigated and hopefully lead to neuroprotective therapies.