TY - JOUR
T1 - Identification of distinct nanoparticles and subsets of extracellular vesicles by asymmetric flow field-flow fractionation
AU - Zhang, Haiying
AU - Freitas, Daniela
AU - Kim, Han Sang
AU - Fabijanic, Kristina
AU - Li, Zhong
AU - Chen, Haiyan
AU - Mark, Milica Tesic
AU - Molina, Henrik
AU - Martin, Alberto Benito
AU - Bojmar, Linda
AU - Fang, Justin
AU - Rampersaud, Sham
AU - Hoshino, Ayuko
AU - Matei, Irina
AU - Kenific, Candia M.
AU - Nakajima, Miho
AU - Mutvei, Anders Peter
AU - Sansone, Pasquale
AU - Buehring, Weston
AU - Wang, Huajuan
AU - Jimenez, Juan Pablo
AU - Cohen-Gould, Leona
AU - Paknejad, Navid
AU - Brendel, Matthew
AU - Manova-Todorova, Katia
AU - Magalhães, Ana
AU - Ferreira, José Alexandre
AU - Osório, Hugo
AU - Silva, André M.
AU - Massey, Ashish
AU - Cubillos-Ruiz, Juan R.
AU - Galletti, Giuseppe
AU - Giannakakou, Paraskevi
AU - Cuervo, Ana Maria
AU - Blenis, John
AU - Schwartz, Robert
AU - Brady, Mary Sue
AU - Peinado, Héctor
AU - Bromberg, Jacqueline
AU - Matsui, Hiroshi
AU - Reis, Celso A.
AU - Lyden, David
N1 - Publisher Copyright:
© 2018 The Author(s).
PY - 2018/3/1
Y1 - 2018/3/1
N2 - 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.
AB - 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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U2 - 10.1038/s41556-018-0040-4
DO - 10.1038/s41556-018-0040-4
M3 - Article
C2 - 29459780
AN - SCOPUS:85042193942
SN - 1465-7392
VL - 20
SP - 332
EP - 343
JO - Nature Cell Biology
JF - Nature Cell Biology
IS - 3
ER -