Characterization of neurogenesis by multidimensional proteomics to develop pure neuronal progenitor cells

Stem cell therapies were suggested to treat incurable neurodegenerative disorders and several clinical trials and/or preclinical studies have been conducted and are ongoing for Parkinson’s disease (PD), amyotrophic lateral sclerosis (ALS), and Huntington’s disease (HD).

However, clinical trials have yielded dispersed outcomes, mostly because human degenerative diseases and neurogenesis are poorly understood.

In this project, we will exploit the high throughput of Proteome Integral Solubility Alteration coupled to expression proteomics (PISA-Express), that I have helped to develop, and couple it with data-independent (DIA) spatial proteomics, DIA phosphoproteomics (Olsen lab, University of Copenhagen) and single-cell proteomics (SCP) to study neurogenesis using four differentiation protocols producing various neuron subtypes.

During years 1 and 2, we will differentiate pluripotent stem cells into the different neuron subtypes, and record protein abundance, thermal stability, subcellular localization, and phosphorylation levels at each key step of the differentiation process. We will use SCP analysis in parallel to evaluate cell populations heterogeneity and study cell fate.

During years 2 and 3 we will compile the data into a publicly available web-based interface that will shed light on protein behavior during neurogenesis.

This will allow us to pinpoint and target key proteins to improve existing differentiation protocols and create purer neuron progenitor cells.