Objective/Rationale:
A-synuclein, the main component of Lewy bodies, is thought to be present mostly on nerve terminals. Although the function of a-synuclein is unclear, its presence in the nucleus of cells has also been reported. The overall objective of the project is to investigate possible effects of a-synuclein in the nucleus of cells and to assess whether it affects the expression of particular genes which may provide clues for therapeutic intervention.
Project Description:
To investigate the effect of a-synuclein on gene expression we will employ state-of-the-art genomic technologies which will enable us to determine the regions in the genome that the protein is binding to, either directly or indirectly. We will employ a technique known as chromatin immunoprecipitation, which will enable us to isolate regions of the genome associated with a-synuclein. Afterwards, sequencing of the DNA fragments will be performed to determine the genes affected. Next, we will use molecular biology techniques to validate our results and to prioritize the hits for future studies in animal models and in patient samples.
Relevance to Diagnosis/Treatment of Parkinson’s Disease:
The molecular mechanisms involved in PD are not fully understood, hindering our ability to intervene therapeutically. Successful identification of genes which are significantly affected by a-synuclein binding could be used in development of other primary assays which may be used in the process of high throughput screening by biotech and pharmaceutical companies, contributing for the development of novel strategies for therapeutic intervention in PD.
Anticipated Outcome:
With this project we expect not only to contribute for the clarification of the function of a-synuclein in the cell, but also to the understanding of the molecular mechanisms involved in its cytotoxicity. The project may provide clues for the discovery of novel targets for therapeutic intervention and may also enable the identification of novel biomarkers for earlier diagnosis.
Final Outcome
The aim of this project was to contribute to the understanding of the role of alpha-synuclein in the nucleus. To this end, we applied a series of advanced post-genomic approaches that enabled us to gain novel insight into the effects of alpha-synuclein. Our results verified the presence of the protein in the nucleus of different types of human cell lines and enabled us to confirm the ability of alpha-synuclein to interact with specific regions of the genome and, therefore, to modulate the expression of a number of genes. In order to further determine the relevance of our findings, we now intend to further study the genes and pathways we identified in order to understand their connection with the molecular mechanisms associated with Parkinson’s. Ultimately, our studies may result in the identification of novel targets for therapeutic intervention in Parkinson’s.