Design and use of EpiEditors to mitigate alpha-synuclein overload in Parkinson’s disease and related synucleinopathies

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Parkinson’s Disease (PD) represents the most common neurodegenerative movement disorder. The hallmark neuropathologic feature of this complex disease is the accumulation of α-synuclein (encoded by SNCA) protein aggregates in Lewy bodies in the PD brain. Rare SNCA point mutations and genomic multiplications are linked to familial PD cases with high penetrance and therefore constitute major genetic risk factors for PD.The key approach of this research proposal is to silence SNCA expression by epigenome editing at the SNCA locus. Thereby, PD brain pathology should be attenuated and cell and tissue homeostasis restored. We will apply advanced epigenome editing systems based on a Sun-Tag and DNMT3A/3L, which will allow very precise, strong, and stable delivery of DNA methylation in different target region in the SNCA regulatory regions. Afterwards, sophisticated cell line and animal models will enable us to investigate the effect of the SNCA silencing on PD symptoms.The specific objectives of this project are to:1. Design and generation of EpiEditors for repression of SNCA expression by targeted DNA methylation.2. Optimization of EpiEditors to maximize locus specific editing, long-term stability of epigenome editing and minimize off-target effects.3. Generation of viral vectors expressing successful EpiEditors.4. Application of human cell culture models to assess the efficacy of EpiEditing in the alleviation of PD-associated pathology at cell culture level.5. Application of EpiEditors in a BAC SNCA mouse and rat model to assess effects of the SNCa promoter methylation on SNCA gene expression, aSYN pathology and behavior.6. Development and application of advanced EpiEditor systems for region specific neuroepiediting with precise temporal control.Successful work in the project depends on the close cooperation of the Jeltsch and Schulze-Hentrich research groups exploiting their complementary and well established expertise in Epigenome editing and investigation of PD.