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Abstract
Extracellular vesicles (EVs) are nanosized (40-1,000nm in diameter) lipid bound vesicles released from most cell types responsible for delivery of functional biologic material to mediate intercellular communication. Physiologically, EVs deliver cargo including RNA, DNA, proteins, and lipids that can functionally modulate recipient cells. Due to their innate characteristics, EV research is a burgeoning field, as therapeutics and delivery vectors of exogenous biomolecules including small interfering RNA, antisense oligonucleotides, and stapled peptides have shown immense promise. EVs possess advantages including low immunogenicity, low toxicity, increased bioavailability, and ability to cross physiological barriers. However, there is limited understanding of EV uptake kinetics and specificity, which may slow therapeutic development. We optimized an imaging flow cytometry platform to assess dose, time, and recipient cell specificity effects on EV internalization. We found that EV uptake is dose and time dependent and that EVs exhibit an inherent specificity to cells of similar origin. To advance the utility of EVs, a novel therapeutic peptide to decrease dopaminergic cell death in Parkinson’s Disease was loaded into EVs via a passive and scalable technique. Herein, EVs can be efficiently loaded with peptides to enhance the peptide uptake kinetics which enabled a more efficient inhibition of the death pathway in Parkinson’s Disease. Lastly, a highly translational porcine stroke model was characterized by identifying lesion topology and assessing its effect on functional outcome post stroke. We found that lesion location impacted stroke recovery as it highly correlated with acute functional outcome. The characterization of lesion topology will advance the understanding of EV therapeutics in the porcine stroke model as a novel mechanism to assess EV biodistribution and function as an alternative biomarker to lesion volume. Together, these studies quantified EV uptake kinetics and specificity, showed that EVs can be harnessed to load with and deliver synthetic peptides, and characterized a translational porcine stroke model to advance the translational potential of extracellular vesicles.