In vitro evaluation of endothelial exosomes as carriers for small interfering ribonucleic acid delivery

In vitro evaluation of endothelial exosomes as carriers for small interfering ribonucleic acid delivery

Anna B Banizs,1 Tao Huang,1 Kelly Dryden,2 Stuart S Berr,1 James R Stone,1 Robert K Nakamoto,2 Weibin Shi,1 Jiang He1 1Department of Radiology and Medical Imaging, 2Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville, VA, USA Abstract: Exoso...

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Autores principales: Banizs AB, Huang T, Dryden K, Berr SS, Stone JR, Nakamoto RK, Shi W, He J
Formato: article
Lenguaje:EN
Publicado: Dove Medical Press 2014
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Medicine (General)
R5-920
Acceso en línea:https://doaj.org/article/4fe899ffaed940dea1a28bb45b36bbaa
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spelling oai:doaj.org-article:4fe899ffaed940dea1a28bb45b36bbaa2021-12-02T00:38:53ZIn vitro evaluation of endothelial exosomes as carriers for small interfering ribonucleic acid delivery1178-2013https://doaj.org/article/4fe899ffaed940dea1a28bb45b36bbaa2014-09-01T00:00:00Zhttp://www.dovepress.com/in-vitro-evaluation-of-endothelial-exosomes-asnbspcarriers-for-small-i-peer-reviewed-article-IJNhttps://doaj.org/toc/1178-2013 Anna B Banizs,1 Tao Huang,1 Kelly Dryden,2 Stuart S Berr,1 James R Stone,1 Robert K Nakamoto,2 Weibin Shi,1 Jiang He1 1Department of Radiology and Medical Imaging, 2Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville, VA, USA Abstract: Exosomes, one subpopulation of nanosize extracellular vesicles derived from multivesicular bodies, ranging from 30 to 150 nm in size, emerged as promising carriers for small interfering ribonucleic acid (siRNA) delivery, as they are capable of transmitting molecular messages between cells through carried small noncoding RNAs, messenger RNAs, deoxyribonucleic acids, and proteins. Endothelial cells are involved in a number of important biological processes, and are a major source of circulating exosomes. In this study, we prepared exosomes from endothelial cells and evaluated their capacity to deliver siRNA into primary endothelial cells. Exosomes were isolated and purified by sequential centrifugation and ultracentrifugation from cultured mouse aortic endothelial cells. Similar to exosome particles from other cell sources, endothelial exosomes are nanometer-size vesicles, examined by both the NanoSight instrument and transmission electron microscopy. Enzyme-linked immunosorbent assay analysis confirmed the expression of two exosome markers: CD9 and CD63. Flow cytometry and fluorescence microscopy studies demonstrated that endothelial exosomes were heterogeneously distributed within cells. In a gene-silencing study with luciferase-expressing endothelial cells, exosomes loaded with siRNA inhibited luciferase expression by more than 40%. In contrast, siRNA alone and control siRNA only suppressed luciferase expression by less than 15%. In conclusion, we demonstrated that endothelial exosomes have the capability to accommodate and deliver short foreign nucleic acids into endothelial cells. Keywords: extracellular vesicles, exosomes, gene delivery, siRNA, endotheliumBanizs ABHuang TDryden KBerr SSStone JRNakamoto RKShi WHe JDove Medical PressarticleMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol 2014, Iss Issue 1, Pp 4223-4230 (2014)
institution DOAJ
collection DOAJ
language EN
topic Medicine (General)
R5-920
spellingShingle Medicine (General)
R5-920
Banizs AB
Huang T
Dryden K
Berr SS
Stone JR
Nakamoto RK
Shi W
He J
In vitro evaluation of endothelial exosomes as carriers for small interfering ribonucleic acid delivery
description Anna B Banizs,1 Tao Huang,1 Kelly Dryden,2 Stuart S Berr,1 James R Stone,1 Robert K Nakamoto,2 Weibin Shi,1 Jiang He1 1Department of Radiology and Medical Imaging, 2Department of Molecular Physiology and Biological Physics, University of Virginia, Charlottesville, VA, USA Abstract: Exosomes, one subpopulation of nanosize extracellular vesicles derived from multivesicular bodies, ranging from 30 to 150 nm in size, emerged as promising carriers for small interfering ribonucleic acid (siRNA) delivery, as they are capable of transmitting molecular messages between cells through carried small noncoding RNAs, messenger RNAs, deoxyribonucleic acids, and proteins. Endothelial cells are involved in a number of important biological processes, and are a major source of circulating exosomes. In this study, we prepared exosomes from endothelial cells and evaluated their capacity to deliver siRNA into primary endothelial cells. Exosomes were isolated and purified by sequential centrifugation and ultracentrifugation from cultured mouse aortic endothelial cells. Similar to exosome particles from other cell sources, endothelial exosomes are nanometer-size vesicles, examined by both the NanoSight instrument and transmission electron microscopy. Enzyme-linked immunosorbent assay analysis confirmed the expression of two exosome markers: CD9 and CD63. Flow cytometry and fluorescence microscopy studies demonstrated that endothelial exosomes were heterogeneously distributed within cells. In a gene-silencing study with luciferase-expressing endothelial cells, exosomes loaded with siRNA inhibited luciferase expression by more than 40%. In contrast, siRNA alone and control siRNA only suppressed luciferase expression by less than 15%. In conclusion, we demonstrated that endothelial exosomes have the capability to accommodate and deliver short foreign nucleic acids into endothelial cells. Keywords: extracellular vesicles, exosomes, gene delivery, siRNA, endothelium
format article
author Banizs AB
Huang T
Dryden K
Berr SS
Stone JR
Nakamoto RK
Shi W
He J
author_facet Banizs AB
Huang T
Dryden K
Berr SS
Stone JR
Nakamoto RK
Shi W
He J
author_sort Banizs AB
title In vitro evaluation of endothelial exosomes as carriers for small interfering ribonucleic acid delivery
title_short In vitro evaluation of endothelial exosomes as carriers for small interfering ribonucleic acid delivery
title_full In vitro evaluation of endothelial exosomes as carriers for small interfering ribonucleic acid delivery
title_fullStr In vitro evaluation of endothelial exosomes as carriers for small interfering ribonucleic acid delivery
title_full_unstemmed In vitro evaluation of endothelial exosomes as carriers for small interfering ribonucleic acid delivery
title_sort in vitro evaluation of endothelial exosomes as carriers for small interfering ribonucleic acid delivery
publisher Dove Medical Press
publishDate 2014
url https://doaj.org/article/4fe899ffaed940dea1a28bb45b36bbaa
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