Investigating the effect of age-related stress on neuron-astrocyte interactions

In this PhD project, we aim to set up a brain co-culture model that would test how astrocyte-to-neuron signalling respond to proteasome deficiency and oxidative stress using primary rodent brain and/or spinal cord, as well as i-PSC derived human cells. Firstly, we aim to characterise neuron-astrocyte interactions using this brain co-culture model and construct a comprehensive secretome analysis with the focus on the characterisation of extracellular vesicles (EVs) and cytokines. Global interactions between neurons and astrocytes will be quantitatively measured using RNA sequencing, cytokine profiling, proteomic mass spectrometry and metabolomic MR spectroscopy. These data will be correlated to normal neuron-astrocyte functions and help us to define biomarkers of healthy neuron-astrocyte interactions. Molecular and cellular assessments of neurons and astrocytes will be performed to further scrutinise the effect of proteasome deficiency in neurons on relevant pathways in reactive astrocytes (e.g., transcriptional regulation, metabolic stress, etc...) and neuron-astrocyte communication (e.g., neuronal death, axon length and glutamate uptake in astrocytes). Finally, we will investigate whether EVs or cytokines released by neurons under age-related stress can control astrocyte reactivity and whether this be targeted to understand fundamental biochemical processes and how they are impacted by ageing and age-associated diseases.