Cited by 32Extracellular vesicles in diseaseMicroRNA in disease regulationNanoparticle-Based Drug Delivery

Differential RNA packaging into small extracellular vesicles by neurons and astrocytes

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Luo, Xuan, Jean-Toussaint, Renée, Sacan, Ahmet, Ajit, Seena K.·2021·Cell Communication and Signaling

Abstract

Background: Small extracellular vesicles (sEVs) mediate intercellular communication by transferring RNA, proteins, and lipids to recipient cells. These cargo molecules are selectively loaded into sEVs and mirror the physiological state of the donor cells. Given that sEVs can cross the blood–brain barrier and their composition can change in neurological disorders, the molecular signatures of sEVs in circulation can be potential disease biomarkers. Characterizing the molecular composition of sEVs from different cell types is an important first step in determining which donor cells contribute to the circulating sEVs. Methods: Cell culture supernatants from primary mouse cortical neurons and astrocytes were used to purify sEVs by differential ultracentrifugation and sEVs were characterized using nanoparticle tracking analysis, transmission electron microscopy and western blot. RNA sequencing was used to determine differential expression and loading patterns of miRNAs in sEVs released by primary neurons and astrocytes. Motif analysis was conducted on enriched miRNAs in sEVs and their respective donor cells. Results: Sequencing total cellular RNA, and miRNAs from sEVs isolated from culture media of postnatal mouse cortical neurons and astrocytes revealed a distinct profile between sEVs and their corresponding cells. Though the total number of detected miRNAs in astrocytes was greater than neurons, neurons expressed more sEV-associated miRNAs than astrocytes. Only 20.7% of astrocytic miRNAs were loaded into sEVs, while 41.0% of neuronal miRNAs were loaded into sEVs, suggesting differences in the cellular sorting mechanisms. We identified short RNA sequence motifs, or EXOmotifs, on the miRNAs that were differentially loaded or excluded from sEVs. A sequence motif GUAC was enriched in astrocytic sEVs. miRNAs preferably retained in neurons or astrocytes had a similar RNA motif CACACA, suggesting a cell-type-independent mechanism to maintain cellular miRNAs. mRNAs of five RNA-binding proteins associated with passive or active RNA sorting into sEVs were differentially expressed between neurons and astrocytes, one of which, major vault protein was higher in astrocytes than in neurons and detected in astrocytic sEVs. Conclusions: Our studies suggest differences in RNA sorting into sEVs. These differences in miRNA signatures can be used for determining the cellular sources of sEVs altered in neurological disorders.

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Authors
Luo, X., Jean-Toussaint, R., Sacan, A., & Ajit, S. K.
Affiliations
Drexel University (US)
Corresponding author
Ajit, Seena K.
Venue
Cell Communication and Signaling
Published
2021-07-10
Volume
19
Issue
1
Pages
75-75
ISSN
1478-811X (ISSN-L)
OpenAlex
W3178242790
Cited by
32
References
78
Citations by year
2026: 7 · 2025: 4 · 2024: 7 · 2023: 10 · 2022: 4
FWCI
1.50
Type
journal article
Language
en
Open access
Locations
  • Cell Communication and Signaling (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
National Institute of Neurological Disorders and Stroke (RO1 NS102836)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › MicroRNA in disease regulation
  • Physical Sciences › Materials Science › Biomaterials › Nanoparticle-Based Drug Delivery
Keywords
small extracellular vesicles, RNA packaging, microRNA, neurons, astrocytes, major vault protein, RNA-binding proteins, intercellular communication, blood-brain barrier, disease biomarkers, cortical neurons, mice
MeSH terms
Extracellular Vesicles, Animals, Astrocytes, Cell Communication, Humans, Neurons, RNA, MicroRNAs, Mice, Nucleotide Motifs

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