Tetraspanins are unevenly distributed across single extracellular vesicles and bias sensitivity to multiplexed cancer biomarkers.
ExoView
Exosomes
Ovarian cancer
Placental mesenchymal stem cells
Single-particle characterization
Journal
Journal of nanobiotechnology
ISSN: 1477-3155
Titre abrégé: J Nanobiotechnology
Pays: England
ID NLM: 101152208
Informations de publication
Date de publication:
21 Aug 2021
21 Aug 2021
Historique:
received:
28
05
2021
accepted:
04
08
2021
entrez:
22
8
2021
pubmed:
23
8
2021
medline:
19
1
2022
Statut:
epublish
Résumé
Tetraspanin expression of extracellular vesicles (EVs) is often used as a surrogate for their detection and classification, a practice that typically assumes their consistent expression across EV sources. Here we demonstrate that there are distinct patterns in colocalization of tetraspanin expression of EVs enriched from a variety of in vitro and in vivo sources. We report an optimized method for the use of single particle antibody-capture and fluorescence detection to identify subpopulations according to tetraspanin expression and compare our findings with nanoscale flow cytometry. We found that tetraspanin profile is consistent from a given EV source regardless of isolation method, but that tetraspanin profiles are distinct across various sources. Tetraspanin profiles measured by flow cytometry do not totally agree, suggesting that limitations in subpopulation detection significantly impact apparent protein expression. We further analyzed tetraspanin expression of single EVs captured non-specifically, revealing that tetraspanin capture can bias the apparent multiplexed tetraspanin profile. Finally, we demonstrate that this bias can have significant impact on diagnostic sensitivity for tumor-associated EV surface markers. Our findings may reveal key insights into protein expression heterogeneity of EVs that better inform EV capture and detection platforms for diagnostic or other downstream use.
Sections du résumé
BACKGROUND
BACKGROUND
Tetraspanin expression of extracellular vesicles (EVs) is often used as a surrogate for their detection and classification, a practice that typically assumes their consistent expression across EV sources.
RESULTS
RESULTS
Here we demonstrate that there are distinct patterns in colocalization of tetraspanin expression of EVs enriched from a variety of in vitro and in vivo sources. We report an optimized method for the use of single particle antibody-capture and fluorescence detection to identify subpopulations according to tetraspanin expression and compare our findings with nanoscale flow cytometry. We found that tetraspanin profile is consistent from a given EV source regardless of isolation method, but that tetraspanin profiles are distinct across various sources. Tetraspanin profiles measured by flow cytometry do not totally agree, suggesting that limitations in subpopulation detection significantly impact apparent protein expression. We further analyzed tetraspanin expression of single EVs captured non-specifically, revealing that tetraspanin capture can bias the apparent multiplexed tetraspanin profile. Finally, we demonstrate that this bias can have significant impact on diagnostic sensitivity for tumor-associated EV surface markers.
CONCLUSION
CONCLUSIONS
Our findings may reveal key insights into protein expression heterogeneity of EVs that better inform EV capture and detection platforms for diagnostic or other downstream use.
Identifiants
pubmed: 34419056
doi: 10.1186/s12951-021-00987-1
pii: 10.1186/s12951-021-00987-1
pmc: PMC8379740
doi:
Substances chimiques
Biomarkers, Tumor
0
Tetraspanins
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
250Subventions
Organisme : NINDS NIH HHS
ID : R01 NS115860
Pays : United States
Organisme : NINDS NIH HHS
ID : R01 NS100761
Pays : United States
Organisme : NIH HHS
ID : T32 OD017863
Pays : United States
Organisme : NCI NIH HHS
ID : P30CA093373
Pays : United States
Organisme : NCI NIH HHS
ID : 1R01CA241666
Pays : United States
Organisme : NCI NIH HHS
ID : R01 CA241666
Pays : United States
Organisme : NCI NIH HHS
ID : P30 CA093373
Pays : United States
Informations de copyright
© 2021. The Author(s).
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