Identification of distinct nanoparticles and subsets of extracellular vesicles by asymmetric flow field-flow fractionation.

TitleIdentification of distinct nanoparticles and subsets of extracellular vesicles by asymmetric flow field-flow fractionation.
Publication TypeJournal Article
Year of Publication2018
AuthorsZhang H, Freitas D, Kim HSang, Fabijanic K, Li Z, Chen H, Mark MTesic, Molina H, Martin ABenito, Bojmar L, Fang J, Rampersaud S, Hoshino A, Matei IR, Kenific CM, Nakajima M, Mutvei APeter, Sansone P, Buehring W, Wang H, Jimenez JPablo, Cohen-Gould L, Paknejad N, Brendel M, Manova-Todorova K, Magalhães A, Ferreira JAlexandre, Osório H, Silva AM, Massey A, Cubillos-Ruiz JR, Galletti G, Giannakakou P, Cuervo AMaria, Blenis J, Schwartz R, Brady MSue, Peinado H, Bromberg JF, Matsui H, Reis CA, Lyden DC
JournalNat Cell Biol
Date Published2018 03
KeywordsAnimals, Biomarkers, Cell Fractionation, DNA, Energy Metabolism, Exosomes, Female, Glycomics, Glycosylation, HCT116 Cells, Humans, Melanoma, Experimental, Metabolomics, Mice, Mice, Inbred C57BL, Nanoparticles, Neoplasms, NIH 3T3 Cells, PC-3 Cells, Phenotype, Proteins, Proteomics, RNA, Signal Transduction, Tissue Distribution

The heterogeneity of exosomal populations has hindered our understanding of their biogenesis, molecular composition, biodistribution and functions. By employing asymmetric flow field-flow fractionation (AF4), we identified two exosome subpopulations (large exosome vesicles, Exo-L, 90-120 nm; small exosome vesicles, Exo-S, 60-80 nm) and discovered an abundant population of non-membranous nanoparticles termed 'exomeres' (~35 nm). Exomere proteomic profiling revealed an enrichment in metabolic enzymes and hypoxia, microtubule and coagulation proteins as well as specific pathways, such as glycolysis and mTOR signalling. Exo-S and Exo-L contained proteins involved in endosomal function and secretion pathways, and mitotic spindle and IL-2/STAT5 signalling pathways, respectively. Exo-S, Exo-L and exomeres each had unique N-glycosylation, protein, lipid, DNA and RNA profiles and biophysical properties. These three nanoparticle subsets demonstrated diverse organ biodistribution patterns, suggesting distinct biological functions. This study demonstrates that AF4 can serve as an improved analytical tool for isolating extracellular vesicles and addressing the complexities of heterogeneous nanoparticle subpopulations.

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Alternate JournalNat. Cell Biol.
PubMed ID29459780
PubMed Central IDPMC5931706
Grant ListT32 CA062948 / CA / NCI NIH HHS / United States
R01 CA218513 / CA / NCI NIH HHS / United States
G12 MD007599 / MD / NIMHD NIH HHS / United States
R01 CA169416 / CA / NCI NIH HHS / United States
U01 CA169538 / CA / NCI NIH HHS / United States
UL1 TR002384 / TR / NCATS NIH HHS / United States
U01 CA210240 / CA / NCI NIH HHS / United States
UL1 TR000457 / TR / NCATS NIH HHS / United States

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