Extracellular vesicle fusion: How is cargo released into target cells.
This project aims to visualize and decipher molecular mechanisms that govern the transfer of extracellular vesicles (EV) between immune cells.
EV are produced by many if not all cells and defined as membrane enclosed cell fragments that are released into the extracellular milieu. Many body fluids contain EV and their cargo is highly pleiotropic (targeting factors, lipids, genetic material and proteins) and its packing is dependent on the status of the donor cells. This is the reason why analyzing molecular profiles of EV isolates is highly suited to monitor pathological development or therapeutic effects.
EV transfer has become recognized as a mode of communication between cells that is unique in the complexity of signals transferred within a single biological unit. Roles for EV in many biological processes have been described mainly due to functional outcome assays that are altered by adding EV isolates. Although the potential of EV function is explained by these studies, molecular regulation mechanisms remain poorly addressed and thus limit optimization for its clinical application.
This project aims to understand mechanisms that control fusion of EV with target cells to release their content. Live cell fluorescence microscopy will be used to track labeled EV in target cells and selectively highlight fusion events. Available donor cell lines with different Fluorescent marker protein fusions allows determination of EV subsets that are effectively targeted. Systematical combination of donor-target cells permits classification of compatible molecular backgrounds for EV transfer. Pharmaceutical and molecular biology studies will be performed to identify essential factors in the process(es).
(Primary) Cell culture, transfection, viral transduction, live cell fluorescence microscopy (TIRF, CLSM), FACS, molecular biology, ultracentrifugation, Super resolution microscopy (SIM/SMLM).
Please contact us via E: R.Wubbolts@uu.nl
Utrecht University, Veterinary Medicine
Department of Biochemistry and Cell Biology
3584 CL Utrecht