Single Particle Characterization and Isolation of Extracellular Vesicles

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Extracellular vesicles (EVs) are submicron-sized biological particles secreted by all forms of life such as cells, bacteria and yeast. EVs carry as cargo molecules from the cells from which they were released (e.g., proteins, lipids, nucleic acids). These can in turn exert biological effects in cells that uptake them such as inducing growth, proliferation, migration and even the secretion of soluble bioactive factors. EVs are released in instances of stress, damage and disease, but can also be released as a means of general communication between biological entities to accomplish a specific task. Proteins and markers at the surface of individual EVs guide them towards their intended target cells that will then engulf them. In complex environments such as bacterial biofilms and biological fluids, EVs from all the cells in the system are released together into the extracellular environment. Because EVs share nearly identical biophysical properties, current purification methods isolate EVs as a bulk. It is therefore nearly impossible to study EV diversity or isolate EVs emanating from specific cells in the system. As such, very little is known about EV diversity, subpopulations and how they impact the behavior of recipient cells in complex and heterogeneous systems. ******My research program aims to understand the function of EVs and how they alter cells in complex environments. To achieve this goal, I propose to:****** i) Establish a new technological analysis platform called NanoFlow that enables single particle characterization and isolation. *** i) Profile surface proteins and lipids, and characterize the diversity of EVs secreted in various complex environments and biological fluids. *** iii) Isolate specific EV populations by NanoFlow sorting, analyze their protein, lipid and nucleic acid content, and determine their biological effects on recipient cells.******The insight provided by detailed EV characterizations will help to better understand how EVs are used to communicate information between cells and coordinate biological effects. Furthermore, the specialized nature of the technology that will be developed by my research program will provide trainees with a unique set of translatable and marketable skills that will no doubt be useful for their future careers in academic research and industry.