Functional Delivery of Lipid-Conjugated siRNA by Extracellular Vesicles

• Published in: Molecular therapy Vol. 25; no. 7; pp. 1580 - 1587
• Main Authors: O’Loughlin, Aisling J., Mäger, Imre, de Jong, Olivier G., Varela, Miguel A., Schiffelers, Raymond M., El Andaloussi, Samir, Wood, Matthew J.A., Vader, Pieter
• Format: Journal Article Conference Proceeding
• Language: English
• Published: United States Elsevier Inc 05-07-2017; Elsevier Limited; American Society of Gene & Cell Therapy
• Subjects: Animals; Base Sequence; Biological Transport; Cancer; Cell interactions; Cell Line; Cell Line, Tumor; Cell signaling; Cell- och molekylärbiologi; Cholesterol; Cholesterol - chemistry; Cholesterol - metabolism; delivery; Drug Delivery Systems; Drug development; Drug dosages; ELAV-Like Protein 1 - antagonists & inhibitors; ELAV-Like Protein 1 - genetics; ELAV-Like Protein 1 - metabolism; exosomes; Extracellular vesicles; Extracellular Vesicles - chemistry; Extracellular Vesicles - metabolism; Fibroblasts - cytology; Fibroblasts - metabolism; Gene Expression; HEK293 Cells; Humans; HuR protein; Medicin och hälsovetenskap; Medicinsk bioteknologi; Medicinsk bioteknologi (inriktn. mot cellbiologi (inkl. stamcellsbiologi), molekylärbiologi, mikrobiologi, biokemi eller biofarmaci); Medicinska och farmaceutiska grundvetenskaper; Mice; Nanoparticles; Neoplasm Proteins - antagonists & inhibitors; Neoplasm Proteins - genetics; Neoplasm Proteins - metabolism; Neurons - metabolism; Neurons - pathology; Original; Permeability; Retention; RNA interference; RNA, Small Interfering - genetics; RNA, Small Interfering - metabolism; siRNA; Temperature; Therapeutic applications; delivery; siRNA; RNA interference; extracellular vesicles; exosomes
• ISSN: 1525-0016; 1525-0024; 1525-0024
• DOI: 10.1016/j.ymthe.2017.03.021

Extracellular vesicles (EVs) are cell-derived, membranous nanoparticles that mediate intercellular communication by transferring biomolecules, including proteins and RNA, between cells. As a result of their suggested natural capability to functionally deliver RNA, EVs may be harnessed as therapeutic RNA carriers. One major limitation for their translation to therapeutic use is the lack of an efficient, robust, and scalable method to load EVs with RNA molecules of interest. Here, we evaluated and optimized methods to load EVs with cholesterol-conjugated small interfering RNAs (cc-siRNAs) by systematic evaluation of the influence of key parameters, including incubation time, volume, temperature, and EV:cc-siRNA ratio. EV loading under conditions that resulted in the highest siRNA retention percentage, incubating 15 molecules of cc-siRNA per EV at 37°C for 1 hr in 100 μL, facilitated concentration-dependent silencing of human antigen R (HuR), a therapeutic target in cancer, in EV-treated cells. These results may accelerate the development of EV-based therapeutics.
Extracellular vesicles (EVs) are cell-derived nanoparticles that mediate intercellular communication. Vader and colleagues demonstrate that EVs can be loaded with cholesterol-conjugated small interfering RNAs, which facilitated knockdown of HuR, a therapeutic target in cancer, in recipient cells. These results may accelerate the development of EV-based therapeutics.