HETLEWY · The Heterogeneity of Lewy Bodies in Single Cells

Parkinson’s Disease (PD) is the second most common neurodegenerative disease affecting millions world-wide. PD pathophysiology is associated with protein aggregates called Lewy Bodies (LBs), and aggregation is impacted by cellular environment, post-translational modifications and genetic predispositions. Heterogeneity in the composition of LBs across the brain has consequences for neuronal cytotoxicity and although LBs are a main focus in PD research, only a limited number of projects have explored their heterogeneity. I will investigate LB heterogeneity at the single cell level for the first time -- enabled by technological developments of my host lab, the Mann group, namely Deep Visual Proteomics (DVP). DVP combines high-resolution microscopy, AI-aided image analysis and ultra-sensitive MS-based proteomics to analyse the proteome in a spatially-resolved manner. I aim to identify proteins whose levels correlate with LB heterogeneity, thereby classifying different LB phenotypes and finding ‘tipping points’ in LB maturation that drive cell death. I will first apply DVP to cell culture models overexpressing alpha-synuclein, a main component of LBs, to investigate the composition of LBs and determine candidates promoting various types of LB phenotypes. Such candidates, e.g., the already known tubulin polymerisation promoting protein (TPPP/p25), will be expressed in cells to confirm effects on the LB phenotype. In parallel, I will apply DVP to human brain tissue to identify differences in aggregates across stages of the disease and to distinguish between healthy and sick cells within the same tissue.This analysis will provide unique insights into the progression of the disease and may yield new therapeutic candidates.