Project - Toll-like receptor 2-mediated α-synuclein release in Parkinson’s disease

Disease area:

PD

Research Project Objectives:

• To determine if activation of neuronal Toll-like receptor 2 (TLR2) cause the release of α-synuclein monomers/aggregates
• To determine if α-synuclein monomers/aggregates are secreted by cells through extracellular vesicles (EVs) after the activation of neuronal TLR2
• To determine if promoting autophagy or pharmacological inhibition of neuronal TLR2 can prevent from the TLR2-mediated α-synuclein pathology/release

Research Project Description

This project is expected to take 3 years (2020-2023).

Background

Parkinson’s disease (PD) is a debilitating and currently incurable neurodegenerative disease, characterized by a loss of dopamine producing neural cells and accumulation of alpha-synuclein protein in the brain. Despite decades of research, exactly how the alpha-synuclein protein accumulates and propagates in PD brain remains unclear. Recently, work from the Dzamko group has shown that activation of toll-like receptor 2 (TLR2) causes alpha-synuclein to accumulate in neurons. TLRs are major regulators of the innate immune inflammatory response, which is normally mediated by specialist immune cells. That TLR2 was expressed and active on neurons was unexpected, but this discovery now opens up new areas of investigation into the role of TLR2 in PD pathogenesis. My project will continue this area of investigation and aims to answer a number of questions that will provide better understanding of the association between neuronal TLR2 activation and the alpha-synuclein pathology that underlies PD.

Methodology

I propose to activate TLR2 in wild type and alpha-synuclein knockout SHSY5Y neuroblastoma cells. I will then collect tissue culture media and detect the presence of alpha-synuclein by immunoblotting/ELISA/dot blot. I will also isolate the EVs in the media using ultra-centrifugation. The isolated EVs will then be analyzed using nanoparticle tracking analysis and immunoblotting to confirm the number and size of EVs isolated. After that, I will use immunoblotting/ELISA/dot blot to measure alpha-synuclein in the EVs.