Chief among the EV subtypes that have captured the interest of clinical researchers are exosomes, which are small (< 200 nm) EVs that begin as the intraluminal vesicles of the late stage endosome, where they are loaded with active biological molecules such as microRNAs (miRNA), mRNA, and proteins ( 3). Our method relies upon established laboratory techniques, can be tailored to a variety of biological questions regarding the pharmacokinetic disposition of extracellular vesicles, and will provide a complementary approach for the of study EV ligand-receptor interactions in the context of EV uptake and targeted therapeutics.Įxtracellular vesicles (EV) can be used to improve medical treatments if properly understood ( 1, 2). Covariate and bootstrap analyses identified cell type having an influence on peripheral volumes (V2 and V3) and clearance (Cl3). We performed an observation-based simulated posterior predictive evaluation with prediction-corrected visual predictive check. Data were best described by a three-compartment model with one elimination from the central compartment. Incorporated oligonucleotide was stable in blood and detectable over five half-lives. ResultsĨ6.5% ± 1.5% (mean ± S.E.) of EV particles were in the 45–195 nm size range and demonstrated protein and lipid markers of endosomal origin. Non-linear mixed effects analysis with first order conditional estimation – extended least squares (FOCE ELS) was used to estimate population-level parameters with associated intra-animal variability. Digital PCR was leveraged to allow for quantification over a wide dynamic range. Jugular vein catheters were used to introduce EVs to conscious rats (n = 30) and to collect blood samples. MethodsĬrude sEVs were labeled with a non-homologous oligonucleotide and isolated from cell culture media using a commercial reagent. Using labeled sEVs administered to conscious rats, we developed a multiple compartment pharmacokinetic model to identify potential differences in the disposition of sEVs from three different cell types. We developed an accessible method for labeling small extracellular vesicles (sEVs) without disrupting endogenous ligands.
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