title: Functional Delivery of Lipid-Conjugated siRNA by Extracellular Vesicles.
abstract: 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.
authors.affiliation: Department of Physiology, Anatomy and Genetics, University of Oxford, OX1 3QX Oxford, UK.
authors.forename: Aisling J
authors.initials: AJ
authors.lastname: O'Loughlin
authors.affiliation: Department of Physiology, Anatomy and Genetics, University of Oxford, OX1 3QX Oxford, UK; Institute of Technology, University of Tartu, Tartu 50411, Estonia.
authors.forename: Imre
authors.initials: I
authors.lastname: Mäger
authors.affiliation: Department of Physiology, Anatomy and Genetics, University of Oxford, OX1 3QX Oxford, UK.
authors.forename: Olivier G
authors.initials: OG
authors.lastname: de Jong
authors.affiliation: Department of Physiology, Anatomy and Genetics, University of Oxford, OX1 3QX Oxford, UK.
authors.forename: Miguel A
authors.initials: MA
authors.lastname: Varela
authors.affiliation: Department of Clinical Chemistry and Haematology, University Medical Center Utrecht, 3584 CX Utrecht, the Netherlands.
authors.forename: Raymond M
authors.initials: RM
authors.lastname: Schiffelers
authors.affiliation: Department of Physiology, Anatomy and Genetics, University of Oxford, OX1 3QX Oxford, UK; Department of Laboratory Medicine, Clinical Research Center, Karolinska Institutet, SE-141 57 Stockholm, Sweden.
authors.forename: Samir
authors.initials: S
authors.lastname: El Andaloussi
authors.affiliation: Department of Physiology, Anatomy and Genetics, University of Oxford, OX1 3QX Oxford, UK. Electronic address: matthew.wood@dpag.ox.ac.uk.
authors.forename: Matthew J A
authors.initials: MJA
authors.lastname: Wood
authors.affiliation: Department of Physiology, Anatomy and Genetics, University of Oxford, OX1 3QX Oxford, UK; Department of Clinical Chemistry and Haematology, University Medical Center Utrecht, 3584 CX Utrecht, the Netherlands. Electronic address: pvader@umcutrecht.nl.
authors.forename: Pieter
authors.initials: P
authors.lastname: Vader
mesh_terms: D000818:Animals; D001483:Base Sequence; D001692:Biological Transport; D002460:Cell Line; D045744:Cell Line, Tumor; D002784:Cholesterol / Q000737:chemistry* / Q000378:metabolism; D016503:Drug Delivery Systems; D000067780:ELAV-Like Protein 1 / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D000067128:Extracellular Vesicles / Q000737:chemistry / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D015870:Gene Expression; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D009363:Neoplasm Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D009474:Neurons / Q000378:metabolism / Q000473:pathology; D010539:Permeability; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism*; D013696:Temperature
chemical_list: D000067780:ELAV-Like Protein 1; C098770:ELAVL1 protein, human; D009363:Neoplasm Proteins; D034741:RNA, Small Interfering; D002784:Cholesterol
keywords: RNA interference; delivery; exosomes; extracellular vesicles; siRNA