Applications of cell resealing to reconstitute microRNA loading to extracellular vesicles

Abstract MicroRNAs (miRNAs) are cargo carried by extracellular vesicles (EVs) and are associated with cell–cell interactions. The response to the cellular environment, such as disease states, genetic/metabolic changes, or differences in cell type, highly regulates cargo sorting to EVs. However, morp...

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Autores principales:	Yuki Sonoda, Fumi Kano, Masayuki Murata
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/da424b01a22c446c86f5444ea5fd1cbf
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spelling	oai:doaj.org-article:da424b01a22c446c86f5444ea5fd1cbf2021-12-02T10:44:14ZApplications of cell resealing to reconstitute microRNA loading to extracellular vesicles10.1038/s41598-021-82452-52045-2322https://doaj.org/article/da424b01a22c446c86f5444ea5fd1cbf2021-02-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-82452-5https://doaj.org/toc/2045-2322Abstract MicroRNAs (miRNAs) are cargo carried by extracellular vesicles (EVs) and are associated with cell–cell interactions. The response to the cellular environment, such as disease states, genetic/metabolic changes, or differences in cell type, highly regulates cargo sorting to EVs. However, morphological features during EV formation and secretion involving miRNA loading are unknown. This study developed a new method of EV loading using cell resealing and reconstituted the elementary miRNA-loading processes. Morphology, secretory response, and cellular uptake ability of EVs obtained from intact and resealed HeLa cells were comparable. Exogenously added soluble factors were introduced into multivesicular endosomes (MVEs) and their subsequent secretion to the extracellular region occurred in resealed HeLa cells. In addition, miRNA transport to MVEs and miRNA encapsulation to EVs followed a distinct pathway regulated by RNA-binding proteins, such as Argonaute and Y-box binding protein 1, depending on miRNA types. Our cell-resealing system can analyze disease-specific EVs derived from disease model cells, where pathological cytosol is introduced into cells. Thus, EV formation in resealed cells can be used not only to create a reconstitution system to give mechanistic insight into EV encapsulation but also for applications such as loading various molecules into EVs and identifying disease-specific EV markers.Yuki SonodaFumi KanoMasayuki MurataNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-18 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Yuki Sonoda Fumi Kano Masayuki Murata Applications of cell resealing to reconstitute microRNA loading to extracellular vesicles
description	Abstract MicroRNAs (miRNAs) are cargo carried by extracellular vesicles (EVs) and are associated with cell–cell interactions. The response to the cellular environment, such as disease states, genetic/metabolic changes, or differences in cell type, highly regulates cargo sorting to EVs. However, morphological features during EV formation and secretion involving miRNA loading are unknown. This study developed a new method of EV loading using cell resealing and reconstituted the elementary miRNA-loading processes. Morphology, secretory response, and cellular uptake ability of EVs obtained from intact and resealed HeLa cells were comparable. Exogenously added soluble factors were introduced into multivesicular endosomes (MVEs) and their subsequent secretion to the extracellular region occurred in resealed HeLa cells. In addition, miRNA transport to MVEs and miRNA encapsulation to EVs followed a distinct pathway regulated by RNA-binding proteins, such as Argonaute and Y-box binding protein 1, depending on miRNA types. Our cell-resealing system can analyze disease-specific EVs derived from disease model cells, where pathological cytosol is introduced into cells. Thus, EV formation in resealed cells can be used not only to create a reconstitution system to give mechanistic insight into EV encapsulation but also for applications such as loading various molecules into EVs and identifying disease-specific EV markers.
format	article
author	Yuki Sonoda Fumi Kano Masayuki Murata
author_facet	Yuki Sonoda Fumi Kano Masayuki Murata
author_sort	Yuki Sonoda
title	Applications of cell resealing to reconstitute microRNA loading to extracellular vesicles
title_short	Applications of cell resealing to reconstitute microRNA loading to extracellular vesicles
title_full	Applications of cell resealing to reconstitute microRNA loading to extracellular vesicles
title_fullStr	Applications of cell resealing to reconstitute microRNA loading to extracellular vesicles
title_full_unstemmed	Applications of cell resealing to reconstitute microRNA loading to extracellular vesicles
title_sort	applications of cell resealing to reconstitute microrna loading to extracellular vesicles
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/da424b01a22c446c86f5444ea5fd1cbf
work_keys_str_mv	AT yukisonoda applicationsofcellresealingtoreconstitutemicrornaloadingtoextracellularvesicles AT fumikano applicationsofcellresealingtoreconstitutemicrornaloadingtoextracellularvesicles AT masayukimurata applicationsofcellresealingtoreconstitutemicrornaloadingtoextracellularvesicles
_version_	1718396794396737536

Applications of cell resealing to reconstitute microRNA loading to extracellular vesicles

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