Supervisor: Andrew Devitt
Project Overview: Cell communication is central to the development of multicellular organisms and their effective physiological functioning. One novel mechanism is mediated by extracellular vesicles (EV). These EV are released from cells when viable, stressed or undergoing cell death and they are taken up by recipient cells where they mediate a broad range of effects including the induction of cell death, cell survival and inflammatory responses. Consequently, these EV are now established as contributing to a range of pathological conditions, including those associated with traumatic brain injury, dementia and diseases of the central nervous system (CNS). It is therefore essential that we understand the composition of these EV released from different cells under different conditions. This will enable us to define how these EV are taken up and how they exert their effects (both desirable and undesirable).
This project will characterise EV release from cells of the neurovascular unit and subsequently identify and characterise key factors associated with EVs that mediate their inflammation-controlling function. Using flow cytometry we will define molecules present on the surface of EV as these are the surface molecules that will facilitate EV uptake by recipient cells and may propagate pathology, as seen in dementia. By defining these molecules, it will be possible to modulate EV uptake for therapeutic gain (e.g. via antibody or small molecule inhibition) and will also highlight potential biomarkers for disease.
- To characterise EV release from cells of the neurovascular unit, in isolation or co-culture.
- To identify key proteins, present on EV that mediate EV function in modulating the innate immune response. Lead proteins for study may be selected from our existing large dataset of mass-spectrometry results of the EV proteome.
- To define the function of selected proteins in EV activity. Using a variety of cell biological and molecular approaches, this will assess proteins as key ligands for EV binding, uptake or active function.
Core Techniques: tissue culture for the harvesting of EV; single particle analyses to characterise EV (number and size); live cell imaging to identify the capacity of EV to recruit macrophages; flow cytometry to reveal the EV surface proteome and flow cytometry of macrophages to define their phenotype after treatment with EV.