Synthetic extracellular vesicles: a selective tool for miRNome profiling in a neuronal environment

MicroRNAs play a pivotal role in the development and function of the central nervous system. They are involved in neurogenesis, neural differentiation, dendritic and axonal growth, synapses development and synaptic plasticity, acting as post-transcriptional regulators of gene expression. Dysregulation of miRNAs function can lead to a pathological behaviour and various neurological disorders. Despite their importance, current technologies for dendritic miRNAs isolation display various technical limitations. Recent literature has demonstrated that the single expression of GAG, one of the three polyproteins constitutive of wild type HIV-1 virus, can generate virus-like particles (VLPs) that are able to encapsulate RNA species from host-cells and that can be further engineered with novel functional units. Our group has designed and generated a panel of lentiviral vectors based on GAG for extracellular vesicles production. The aim of my thesis is to validate these novel tools in living cells in a multicellular environment and to isolate and identify post-synaptic miRNAs in healthy and Alzheimer’s disease cellular models.