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Profiling of Extracellular Small RNAs Highlights a Strong Bias towards Non-Vesicular Secretion
The extracellular environment consists of a plethora of molecules, including extracellular miRNA that can be secreted in association with extracellular vesicles (EVs) or soluble protein complexes (non-EVs). Yet, interest in therapeutic short RNA carriers lies mainly in EVs, the vehicles conveying th...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8235078/ https://www.ncbi.nlm.nih.gov/pubmed/34207405 http://dx.doi.org/10.3390/cells10061543 |
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author | Sork, Helena Conceicao, Mariana Corso, Giulia Nordin, Joel Lee, Yi Xin Fiona Krjutskov, Kaarel Orzechowski Westholm, Jakub Vader, Pieter Pauwels, Marie Vandenbroucke, Roosmarijn E. Wood, Matthew JA EL Andaloussi, Samir Mäger, Imre |
author_facet | Sork, Helena Conceicao, Mariana Corso, Giulia Nordin, Joel Lee, Yi Xin Fiona Krjutskov, Kaarel Orzechowski Westholm, Jakub Vader, Pieter Pauwels, Marie Vandenbroucke, Roosmarijn E. Wood, Matthew JA EL Andaloussi, Samir Mäger, Imre |
author_sort | Sork, Helena |
collection | PubMed |
description | The extracellular environment consists of a plethora of molecules, including extracellular miRNA that can be secreted in association with extracellular vesicles (EVs) or soluble protein complexes (non-EVs). Yet, interest in therapeutic short RNA carriers lies mainly in EVs, the vehicles conveying the great majority of the biological activity. Here, by overexpressing miRNA and shRNA sequences in parent cells and using size exclusion liquid chromatography (SEC) to separate the secretome into EV and non-EV fractions, we saw that >98% of overexpressed miRNA was secreted within the non-EV fraction. Furthermore, small RNA sequencing studies of native miRNA transcripts revealed that although the abundance of miRNAs in EVs, non-EVs and parent cells correlated well (R(2) = 0.69–0.87), quantitatively an outstanding 96.2–99.9% of total miRNA was secreted in the non-EV fraction. Nevertheless, though EVs contained only a fraction of secreted miRNAs, these molecules were stable at 37 °C in a serum-containing environment, indicating that if sufficient miRNA loading is achieved, EVs can remain delivery-competent for a prolonged period of time. This study suggests that the passive endogenous EV loading strategy might be a relatively wasteful way of loading miRNA to EVs, and active miRNA loading approaches are needed for developing advanced EV miRNA therapies in the future. |
format | Online Article Text |
id | pubmed-8235078 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-82350782021-06-27 Profiling of Extracellular Small RNAs Highlights a Strong Bias towards Non-Vesicular Secretion Sork, Helena Conceicao, Mariana Corso, Giulia Nordin, Joel Lee, Yi Xin Fiona Krjutskov, Kaarel Orzechowski Westholm, Jakub Vader, Pieter Pauwels, Marie Vandenbroucke, Roosmarijn E. Wood, Matthew JA EL Andaloussi, Samir Mäger, Imre Cells Article The extracellular environment consists of a plethora of molecules, including extracellular miRNA that can be secreted in association with extracellular vesicles (EVs) or soluble protein complexes (non-EVs). Yet, interest in therapeutic short RNA carriers lies mainly in EVs, the vehicles conveying the great majority of the biological activity. Here, by overexpressing miRNA and shRNA sequences in parent cells and using size exclusion liquid chromatography (SEC) to separate the secretome into EV and non-EV fractions, we saw that >98% of overexpressed miRNA was secreted within the non-EV fraction. Furthermore, small RNA sequencing studies of native miRNA transcripts revealed that although the abundance of miRNAs in EVs, non-EVs and parent cells correlated well (R(2) = 0.69–0.87), quantitatively an outstanding 96.2–99.9% of total miRNA was secreted in the non-EV fraction. Nevertheless, though EVs contained only a fraction of secreted miRNAs, these molecules were stable at 37 °C in a serum-containing environment, indicating that if sufficient miRNA loading is achieved, EVs can remain delivery-competent for a prolonged period of time. This study suggests that the passive endogenous EV loading strategy might be a relatively wasteful way of loading miRNA to EVs, and active miRNA loading approaches are needed for developing advanced EV miRNA therapies in the future. MDPI 2021-06-18 /pmc/articles/PMC8235078/ /pubmed/34207405 http://dx.doi.org/10.3390/cells10061543 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Sork, Helena Conceicao, Mariana Corso, Giulia Nordin, Joel Lee, Yi Xin Fiona Krjutskov, Kaarel Orzechowski Westholm, Jakub Vader, Pieter Pauwels, Marie Vandenbroucke, Roosmarijn E. Wood, Matthew JA EL Andaloussi, Samir Mäger, Imre Profiling of Extracellular Small RNAs Highlights a Strong Bias towards Non-Vesicular Secretion |
title | Profiling of Extracellular Small RNAs Highlights a Strong Bias towards Non-Vesicular Secretion |
title_full | Profiling of Extracellular Small RNAs Highlights a Strong Bias towards Non-Vesicular Secretion |
title_fullStr | Profiling of Extracellular Small RNAs Highlights a Strong Bias towards Non-Vesicular Secretion |
title_full_unstemmed | Profiling of Extracellular Small RNAs Highlights a Strong Bias towards Non-Vesicular Secretion |
title_short | Profiling of Extracellular Small RNAs Highlights a Strong Bias towards Non-Vesicular Secretion |
title_sort | profiling of extracellular small rnas highlights a strong bias towards non-vesicular secretion |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8235078/ https://www.ncbi.nlm.nih.gov/pubmed/34207405 http://dx.doi.org/10.3390/cells10061543 |
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