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Delivery of Small Interfering RNA to Inhibit Vascular Endothelial Growth Factor in Zebrafish Using Natural Brain Endothelia Cell-Secreted Exosome Nanovesicles for the Treatment of Brain Cancer

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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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ACKNOWLEDGEMENTS

Research reported in this project was supported by the Husson University Faculty Research Funds and School of Pharmacy Seed Grants.

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Authors and Affiliations

  1. Department of Basic Pharmaceutical Sciences, School of Pharmacy, Husson University, Bangor, Maine, 04401, USA

    Tianzhi Yang, Brittany Fogarty, Bret LaForge, Salma Aziz, Thuy Pham & Shuhua Bai

  2. Department of Sociobehavioral and Administrative Pharmacy, College of Pharmacy, Nova Southeastern University, Ft. Lauderdale, Florida, 33328, USA

    Leanne Lai

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  1. Tianzhi Yang
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  2. Brittany Fogarty
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  3. Bret LaForge
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Correspondence to Shuhua Bai.

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Yang, T., Fogarty, B., LaForge, B. et al. Delivery of Small Interfering RNA to Inhibit Vascular Endothelial Growth Factor in Zebrafish Using Natural Brain Endothelia Cell-Secreted Exosome Nanovesicles for the Treatment of Brain Cancer. AAPS J 19, 475–486 (2017). https://doi.org/10.1208/s12248-016-0015-y

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