Effects of Inhibiting VPS4 Support a General Role for ESCRTs in Extracellular Vesicle Biogenesis
Open accessAbstract
Extracellular vesicles (EVs) are proposed to play important roles in intercellular communication. Two classes of EVs can be distinguished based on their intracellular origin. Exosomes are generated within endosomes and released when these fuse with the plasma membrane, whereas ectosomes bud directly from the plasma membrane. Studies of EV function have been hindered by limited understanding of their biogenesis. Components of the endosomal sorting complex required for transport (ESCRT) machinery play essential roles in topologically equivalent processes at both the endosome and the plasma membrane and are consistently recovered in EVs, but whether they are generally required to produce EVs is still debated. Here, we study the effects of inhibiting the ESCRT-associated AAA+ ATPase VPS4 on EV release from cultured cells using two methods for EV recovery, differential centrifugation and polyethylene glycol precipitation followed by lectin affinity chromatography. We find that inhibiting VPS4 in HEK293 cells decreases release of EV-associated proteins and miRNA as well as the overall number of EV particles. The tetraspanins CD63 and CD9 are among the most frequently monitored EV proteins, but they differ in their subcellular localization, with CD63 primarily in endosomes and CD9 on the plasma membrane. We find that CD63 and CD9 are enriched in separable populations of EVs that are both sensitive to VPS4 inhibition. Serum stimulation increases release of both types of EVs and is also reduced by inhibiting VPS4. Taken together, our data indicate that VPS4 activity is important for generating exosomes and ectosomes, thereby generally implicating the ESCRT machinery in EV biogenesis.
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Vps4 and ALIX participate in cargo sorting and exosome formation.
“In addition, Vps4 and ALIX also participate in cargo sorting and exosome formation [15,16].”
From The Therapeutic Potential and Clinical Significance of Exosomes as Carriers of Drug Delivery System (2022) · cites this paper as [15]
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- Authors
- Jackson, C. E., Scruggs, B. S., Schaffer, J. E., & Hanson, P. I.
- Affiliations
- Washington University in St. Louis (US)
- Corresponding author
- Hanson, Phyllis I.
- Venue
- Biophysical Journal
- Publisher
- Elsevier BV
- Published
- 2017-09
- Volume
- 113
- Issue
- 6
- Pages
- 1342-1352
- ISSN
- 0006-3495 (ISSN-L)
- PMC
- PMC5607042
- OpenAlex
- W2624735470
- Cited by
- 118
- References
- 72
- Citations by year
- 2026: 14 · 2025: 11 · 2024: 16 · 2023: 17 · 2022: 12 · 2021: 14 · 2020: 12 · 2019: 11 · 2018: 9 · 2017: 2
- FWCI
- 3.91
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Biophysical Journal (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Caveolin-1 and cellular processes
- Keywords
- extracellular vesicles, EV biogenesis, ESCRT machinery, VPS4, AAA+ ATPase, exosomes, ectosome, CD63, CD9, tetraspanins, serum stimulation, endosomal sorting
- MeSH terms
- Adenosine Triphosphatases, Humans, Endosomes, Protein Transport, Endosomal Sorting Complexes Required for Transport, HEK293 Cells
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