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DNA analysis of low- and high-density fractions defines heterogeneous subpopulations of small extracellular vesicles based on their DNA cargo and topology

Extracellular vesicles have the capacity to transfer lipids, proteins, and nucleic acids between cells, thereby influencing the recipient cell’s phenotype. While the role of RNAs in EVs has been extensively studied, the function of DNA remains elusive. Here, we distinguished novel heterogeneous subp...

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Autores principales: Lázaro-Ibáñez, Elisa, Lässer, Cecilia, Shelke, Ganesh Vilas, Crescitelli, Rossella, Jang, Su Chul, Cvjetkovic, Aleksander, García-Rodríguez, Anaís, Lötvall, Jan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Taylor & Francis 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6719264/
https://www.ncbi.nlm.nih.gov/pubmed/31497265
http://dx.doi.org/10.1080/20013078.2019.1656993
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author Lázaro-Ibáñez, Elisa
Lässer, Cecilia
Shelke, Ganesh Vilas
Crescitelli, Rossella
Jang, Su Chul
Cvjetkovic, Aleksander
García-Rodríguez, Anaís
Lötvall, Jan
author_facet Lázaro-Ibáñez, Elisa
Lässer, Cecilia
Shelke, Ganesh Vilas
Crescitelli, Rossella
Jang, Su Chul
Cvjetkovic, Aleksander
García-Rodríguez, Anaís
Lötvall, Jan
author_sort Lázaro-Ibáñez, Elisa
collection PubMed
description Extracellular vesicles have the capacity to transfer lipids, proteins, and nucleic acids between cells, thereby influencing the recipient cell’s phenotype. While the role of RNAs in EVs has been extensively studied, the function of DNA remains elusive. Here, we distinguished novel heterogeneous subpopulations of small extracellular vesicles (sEVs) based on their DNA content and topology. Low- and high-density sEV subsets from a human mast cell line (HMC-1) and an erythroleukemic cell line (TF-1) were separated using high-resolution iodixanol density gradients to discriminate the nature of the DNA cargo of the sEVs. Paired comparisons of the sEV-associated DNA and RNA molecules showed that RNA was more abundant than DNA and that most of the DNA was present in the high-density fractions, demonstrating that sEV subpopulations have different DNA content. DNA was predominately localised on the outside or surface of sEVs, with only a small portion being protected from enzymatic degradation. Whole-genome sequencing identified DNA fragments spanning all chromosomes and mitochondrial DNA when sEVs were analysed in bulk. Our work contributes to the understanding of how DNA is associated with sEVs and thus provides direction for distinguishing subtypes of EVs based on their DNA cargo and topology.
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spelling pubmed-67192642019-09-06 DNA analysis of low- and high-density fractions defines heterogeneous subpopulations of small extracellular vesicles based on their DNA cargo and topology Lázaro-Ibáñez, Elisa Lässer, Cecilia Shelke, Ganesh Vilas Crescitelli, Rossella Jang, Su Chul Cvjetkovic, Aleksander García-Rodríguez, Anaís Lötvall, Jan J Extracell Vesicles Research Article Extracellular vesicles have the capacity to transfer lipids, proteins, and nucleic acids between cells, thereby influencing the recipient cell’s phenotype. While the role of RNAs in EVs has been extensively studied, the function of DNA remains elusive. Here, we distinguished novel heterogeneous subpopulations of small extracellular vesicles (sEVs) based on their DNA content and topology. Low- and high-density sEV subsets from a human mast cell line (HMC-1) and an erythroleukemic cell line (TF-1) were separated using high-resolution iodixanol density gradients to discriminate the nature of the DNA cargo of the sEVs. Paired comparisons of the sEV-associated DNA and RNA molecules showed that RNA was more abundant than DNA and that most of the DNA was present in the high-density fractions, demonstrating that sEV subpopulations have different DNA content. DNA was predominately localised on the outside or surface of sEVs, with only a small portion being protected from enzymatic degradation. Whole-genome sequencing identified DNA fragments spanning all chromosomes and mitochondrial DNA when sEVs were analysed in bulk. Our work contributes to the understanding of how DNA is associated with sEVs and thus provides direction for distinguishing subtypes of EVs based on their DNA cargo and topology. Taylor & Francis 2019-08-27 /pmc/articles/PMC6719264/ /pubmed/31497265 http://dx.doi.org/10.1080/20013078.2019.1656993 Text en © 2019 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group on behalf of The International Society for Extracellular Vesicles. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Lázaro-Ibáñez, Elisa
Lässer, Cecilia
Shelke, Ganesh Vilas
Crescitelli, Rossella
Jang, Su Chul
Cvjetkovic, Aleksander
García-Rodríguez, Anaís
Lötvall, Jan
DNA analysis of low- and high-density fractions defines heterogeneous subpopulations of small extracellular vesicles based on their DNA cargo and topology
title DNA analysis of low- and high-density fractions defines heterogeneous subpopulations of small extracellular vesicles based on their DNA cargo and topology
title_full DNA analysis of low- and high-density fractions defines heterogeneous subpopulations of small extracellular vesicles based on their DNA cargo and topology
title_fullStr DNA analysis of low- and high-density fractions defines heterogeneous subpopulations of small extracellular vesicles based on their DNA cargo and topology
title_full_unstemmed DNA analysis of low- and high-density fractions defines heterogeneous subpopulations of small extracellular vesicles based on their DNA cargo and topology
title_short DNA analysis of low- and high-density fractions defines heterogeneous subpopulations of small extracellular vesicles based on their DNA cargo and topology
title_sort dna analysis of low- and high-density fractions defines heterogeneous subpopulations of small extracellular vesicles based on their dna cargo and topology
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6719264/
https://www.ncbi.nlm.nih.gov/pubmed/31497265
http://dx.doi.org/10.1080/20013078.2019.1656993
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