Cellular uptake of extracellular vesicles is mediated by clathrin-independent endocytosis and macropinocytosis

Journal of Controlled Release, Journal Year: 2017, Volume and Issue: 266, P. 100 - 108

Published: Sept. 14, 2017

Recent evidence has established that extracellular vesicles (EVs), including exosomes and microvesicles, form an endogenous transport system through which biomolecules, proteins RNA, are exchanged between cells. This endows EVs with immense potential for drug delivery regenerative medicine applications. Understanding the biology underlying EV-based intercellular transfer of cargo is great importance development therapeutics. Here, we sought to characterize cellular mechanisms involved in EV uptake. Internalization fluorescently-labeled was evaluated HeLa cells, 2D (monolayer) cell culture as well 3D spheroids. Uptake assessed using flow cytometry confocal microscopy, chemical RNA interference-based inhibition key individual endocytic pathways. Experiments inhibitors revealed uptake depends on cholesterol tyrosine kinase activity, implicated clathrin-independent endocytosis, Na+/H+ exchange phosphoinositide 3-kinase important macropinocytosis. Furthermore, internalization inhibited by siRNA-mediated knockdown caveolin-1, flotillin-1, RhoA, Rac1 PAK1, but not clathrin heavy chain. Together, these results suggest enter cells predominantly via endocytosis Identification components promote their pathways lead functional might allow more efficient therapeutics EV-inspired engineering.

Language: Английский

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RNA in extracellular vesicles

Wiley Interdisciplinary Reviews - RNA, Journal Year: 2017, Volume and Issue: 8(4)

Published: Jan. 28, 2017

Cells release a range of membrane‐enclosed extracellular vesicles ( EVs ) into the environment. Among them, exosomes and microvesicles (collectively measuring 40–1000 nm in diameter) carry proteins, signaling lipids, nucleic acids from donor cells to recipient cells, thus have been proposed serve as intercellular mediators communication. transport cellular materials many physiologic processes, including differentiation, stem cell homeostasis, immune responses, neuronal signaling. are also increasingly recognized having direct role pathologies such cancer neurodegeneration. Accordingly, focus intense investigation biomarkers disease, prognostic indicators, even therapeutic tools. Here, we review classes RNAs present , both coding (messenger RNAs) noncoding (long microRNAs circular ). The rising attention EV ‐resident stems fact that can be detected at extremely low quantities using number methods. To illustrate interest biology, discuss neurodegeneration, two major age‐associated disease processes. WIREs RNA 2017, 8:e1413. doi: 10.1002/wrna.1413 This article is categorized under: Export Localization >

Language: Английский

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Asymmetric-flow field-flow fractionation technology for exomere and small extracellular vesicle separation and characterization

Nature Protocols, Journal Year: 2019, Volume and Issue: 14(4), P. 1027 - 1053

Published: March 4, 2019

Language: Английский

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Exosomes in cancer development and clinical applications

Cancer Science, Journal Year: 2018, Volume and Issue: 109(8), P. 2364 - 2374

Published: June 16, 2018

Exosomes participate in cancer progression and metastasis by transferring bioactive molecules between various cells the local distant microenvironments. Such intercellular cross-talk results changes multiple cellular biological functions recipient cells. Several hallmarks of have reportedly been impacted this exosome-mediated cell-to-cell communication, including modulating immune responses, reprogramming stromal cells, remodeling architecture extracellular matrix, or even endowing with characteristics drug resistance. Selectively, loading specific oncogenic into exosomes highlights as potential diagnostic biomarkers well therapeutic targets. In addition, exosome-based delivery strategies preclinical clinical trials shown to dramatically decrease development. present review, we summarize significant aspects development that can provide novel for applications.

Language: Английский

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Isolation and characterization of extracellular vesicle subpopulations from tissues

Nature Protocols, Journal Year: 2021, Volume and Issue: 16(3), P. 1548 - 1580

Published: Jan. 25, 2021

Language: Английский

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