Proteomics of neuroinflammatory signaling: Illuminating cellular mechanisms driving neurodegenerative diseases

Alzheimers (AD) and Parkinsons disease (PD) are prevalent neurodegenerative diseases, but their underlying mechanisms remain poorly understood.

Emerging evidence suggests that neuroinflammation, characterized by the release of pro-inflammatory cytokines like IL-1, TNF-, and INF-, plays a pivotal role in these diseases, challenging the conventional focus on protein aggregates (A, Syn).

In AD, it has been shown that elevated IL-1 levels induce the translocation of NEDD8 from the nucleus to the cytoplasm, where it co-localizes with the E3-ligase Parkin.

Together with the kinase PINK1, Parkin is crucial for the removal of defective mitochondria, with loss-of-function mutations in these genes being the primary cause of early-onset PD.

Intriguingly, phosphorylated NEDD8 can activate Parkin, implicating a more complex interconnection between neuroinflammation, NEDD8-and Parkin-regulated phosphorylation-dependent signaling in brain cells.

Yet, a mechanistic understanding of cellular processes triggered by neuroinflammation remain elusive.This proposal aims to investigate spatiotemporal (phospho-)proteome changes in response to neuroinflammatory stress elicited by pro-inflammatory cytokines and Syn fibrils.

To achieve this, I will combine spatial-proteomics using subcellular fractionation and hybrid-DIA in combination with the cutting-edge Orbitrap Astral mass spectrometer.

Initial analyses of neuronal cell line models will be extended in a second step to hPSC derived neurons, astrocytes, microglia, and co-cultures thereof.

This enhances the translational relevance by mimicking neuroinflammation in the complex tissue architecture of the brain.

Finally, bioinformatically shortlisted neuroinflammation-regulated proteins and phospho-sites will be validated in cell-based models.

With this project, I aspire to unravel the cellular mechanisms underlying neuroinflammation, potentially unlocking new avenues for therapeutic strategies against neurodegenerative diseases.