Abstract |
Although small interfering RNA ( siRNA) holds great therapeutic promise, its delivery to the disease site remains a paramount obstacle. In this study, we tested whether brain endothelial cell-derived exosomes could deliver siRNA across the blood-brain barrier (BBB) in zebrafish. Natural exosomes were isolated from brain endothelial bEND.3 cell culture media and vascular endothelial growth factor ( VEGF) siRNA was loaded in exosomes with the assistance of a transfection reagent. While fluorescence-activated cell flow cytometry and immunocytochemistry staining studies indicated that wild-type exosomes significantly increased the uptake of fluorescence-labeled siRNA in the autologous brain endothelial cells, decreased fluorescence intensity was observed in the cells treated with the tetraspanin CD63 antibody-blocked exosome-delivered formulation (p < 0.05). In the transport study, exosomes also enhanced the permeability of rhodamine 123 in a co-cultured monolayer of brain endothelial bEND.3 cell and astrocyte. Inhibition at the expression of VEGF RNA and protein levels was observed in glioblastoma- astrocytoma U-87 MG cells treated with exosome-delivered siRNAs. Imaging results showed that exosome delivered more siRNAs across the BBB in Tg(fli1:GFP) zebrafish. In a xenotransplanted brain tumor model, exosome-delivered VEGF siRNAs decreased the fluorescence intensity of labeled cancer cells in the brain of zebrafish. Brain endothelial cell-derived exosomes could be potentially used as a natural carrier for the brain delivery of exogenous siRNA.
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Authors | Tianzhi Yang, Brittany Fogarty, Bret LaForge, Salma Aziz, Thuy Pham, Leanne Lai, Shuhua Bai |
Journal | The AAPS journal
(AAPS J)
Vol. 19
Issue 2
Pg. 475-486
(03 2017)
ISSN: 1550-7416 [Electronic] United States |
PMID | 27882487
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- RNA, Small Interfering
- Vascular Endothelial Growth Factor A
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Topics |
- Animals
- Astrocytes
(metabolism)
- Blood-Brain Barrier
(metabolism)
- Brain
(metabolism, pathology)
- Brain Neoplasms
(genetics, pathology, therapy)
- Cell Line, Tumor
- Coculture Techniques
- Endothelial Cells
(metabolism)
- Exosomes
(metabolism)
- Gene Transfer Techniques
- Humans
- RNA, Small Interfering
(administration & dosage)
- Transfection
- Vascular Endothelial Growth Factor A
(antagonists & inhibitors, genetics)
- Zebrafish
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