(P03-013-23) Apple and Orange-Derived Extracellular Vesicles As Nanocarriers of Extracellular RNAs

Objectives: Although extracellular RNA is now known to travel outside cells to tissues around the body, when unprotected it undergoes rapid degradation. As extracellular RNAs can be naturally transported in extracellular vesicles (EVs), the latter have been proposed as low-immunogenic shipping means for drug delivery, granting stability and less susceptibility to degradation by RNases. However, a standard, reproducible, scalable, and cost-effective protocol for EV isolation from natural sources is missing. Here, the goal was to survey the potential of apple- and orange-derived EVs as therapeutic vehicles for extracellular RNA (drug) delivery.

Methods: EVs were isolated from apple and orange, combining ultracentrifugation and size exclusion chromatography. EVs were characterized by means of Nanoparticle Tracking Analysis and electron microscopy. Isolated EVs were loaded with exogenous miRNAs and then assayed for RNase degradation. Cellular uptake of EV-ferried miRNA was tested in the intestinal Caco-2 cell line.

Results: Apple- and orange-derived EVs were efficiently isolated, characterized and loaded with exogenous miRNAs. Fruit-derived EVs further protected extracellular miRNAs from RNase degradation compared to unloaded miRNAs. Extracellular RNAs packed in EVs were efficiently taken up by target cells.

Conclusions: EVs recovered from natural sources such as edible fruits (i.e., apple and orange) are capable of transporting and delivering exogenous miRNAs to target cells and are proposed as potentially important players in the therapeutic arena.

Funding Sources: Spanish Agencia Estatal de Investigación, European FEDER Funds, and Spanish Foundation of Arteriosclerosis (“Manuel de Oya” Nutrition Grant 2021).