Understanding the role of astrocyte-derived extracellular vesicles in health and disease

We aim to construct a comprehensive, multi-omics astrocyte secretome (including RNA/proteins/metabolites) with the focus on the characterisation of extracellular vesicles (EVs). The content of astrocyte EVs will be compared to that of non-EV secreted fraction (i.e. intracellular analytes in cell lysates). We will then determine the mechanism by which astrocyte-derived EVs regulate astrocyte reactivity and neuron-astrocyte communication in health and disease. Broadly relevant to neurodegeneration, we will focus on proteins associated to amyotrophic lateral sclerosis (ALS) (e.g. RNA binding, ubiquitin-binding proteins) and determine to what extend ALS mutations can result in dysfunction of the loading and secretion of EVs. To this end, the effect of ALS will be mimicked using genetic/pharmacological manipulation of autophagy or oxidative stress. Molecular and cellular assessments of neuron and astrocytes will be performed to further scrutinise the effect of dysfunction EVs 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 represent 'metabolically functional units' with the ability to function independently of donor cells, as potential for biomarkers of phenotypic changes and therapeutic target.