Subpopulations of extracellular vesicles from human metastatic melanoma tissue identified by quantitative proteomics after optimized isolation

Journal of Extracellular Vesicles
Rossella CrescitelliJan Lötvall

Abstract

The majority of extracellular vesicle (EV) studies conducted to date have been performed on cell lines with little knowledge on how well these represent the characteristics of EVs in vivo. The aim of this study was to establish a method to isolate and categorize subpopulations of EVs isolated directly from tumour tissue. First we established an isolation protocol for subpopulations of EVs from metastatic melanoma tissue, which included enzymatic treatment (collagenase D and DNase). Small and large EVs were isolated with differential ultracentrifugation, and these were further separated into high and low-density (HD and LD) fractions. All EV subpopulations were then analysed in depth using electron microscopy, Bioanalyzer®, nanoparticle tracking analysis, and quantitative mass spectrometry analysis. Subpopulations of EVs with distinct size, morphology, and RNA and protein cargo could be isolated from the metastatic melanoma tissue. LD EVs showed an RNA profile with the presence of 18S and 28S ribosomal subunits. In contrast, HD EVs had RNA profiles with small or no peaks for ribosomal RNA subunits. Quantitative proteomics showed that several proteins such as flotillin-1 were enriched in both large and small LD EVs, while ADAM10 ...Continue Reading

References

Jan 1, 1996·Journal of Structural Biology·J R KremerJ R McIntosh
Nov 23, 2000·The Journal of Biological Chemistry·C S StippM E Hemler
Mar 30, 2001·The Journal of Biological Chemistry·S CharrinE Rubinstein
Sep 24, 2005·The Journal of Biological Chemistry·Martine GirardPeter S McPherson
Oct 13, 2007·Cell Death and Differentiation·M IeroL Rivoltini
Feb 8, 2008·Cell Structure and Function·Akemi FujibayashiKiyotoshi Sekiguchi
May 29, 2009·Digestive and Liver Disease : Official Journal of the Italian Society of Gastroenterology and the Italian Association for the Study of the Liver·J-L QuY-P Liu
Jul 6, 2010·Journal of Proteomics·Suresh MathivananRichard J Simpson
Jan 18, 2011·Journal of Translational Medicine·Cecilia LässerHadi Valadi
Mar 9, 2011·Proceedings of the National Academy of Sciences of the United States of America·Jason D ArroyoMuneesh Tewari
May 10, 2011·The American Journal of Dermatopathology·Charay Jennings, Jinah Kim
Jun 10, 2011·Journal of Translational Medicine·Sascha KellerPeter Altevogt
Feb 20, 2013·The Journal of Cell Biology·Graça Raposo, Willem Stoorvogel
Jul 11, 2013·Nature Communications·Silvia DomckeNikolaus Schultz
Sep 7, 2013·Journal of Extracellular Vesicles·Dae-Kyum KimYong Song Gho
Nov 14, 2013·Journal of Extracellular Vesicles·Rossella CrescitelliJan Lötvall
Jan 1, 2014·Proceedings of the National Academy of Sciences of the United States of America·Safinur AtayAndrew K Godwin
Jun 1, 2012·European Journal of Microbiology & Immunology·A AutengruberD Bruder
Oct 15, 2014·Cell Communication and Signaling : CCS·Hui XiaoJan Lötvall
Oct 16, 2014·Journal of Extracellular Vesicles·Jan Van DeunAn Hendrix
Oct 6, 2015·Journal of Molecular Biology·Shivakumar KeerthikumarSuresh Mathivanan
Feb 10, 2016·Proceedings of the National Academy of Sciences of the United States of America·Joanna KowalClotilde Théry
Aug 10, 2016·Scientific Reports·Yong Hyun ParkJi Youl Lee
Sep 2, 2016·Science Translational Medicine·Marie AllenBengt Westermark
Oct 21, 2016·The American Journal of Dermatopathology·Oliver Chang, Zsolt Argenyi
Mar 1, 2017·Nature Methods·UNKNOWN EV-TRACK ConsortiumAn Hendrix
Jul 19, 2017·Journal of Extracellular Vesicles·Davide ZabeoJohanna L Höög
Aug 15, 2017·Journal of Extracellular Vesicles·Laura J VellaAndrew F Hill
Dec 14, 2017·Scientific Reports·Kristina SvennerholmJan Lötvall
Feb 13, 2018·Molecular Aspects of Medicine·Cecilia LässerJan Lötvall
Apr 6, 2019·Cell·Dennis K JeppesenRobert J Coffey

❮ Previous
Next ❯

Citations

Aug 1, 2020·International Journal of Molecular Sciences·Kerstin MenckClaudia Binder
Jul 1, 2020·Journal of Extracellular Vesicles·Shiv Ram KrishnLucia R Languino
Jul 29, 2020·Cell Death & Disease·Ethan Z Malkin, Scott V Bratman
Dec 19, 2020·Translational Oncology·Zarema AlbakovaAlexander M Sapozhnikov
Jan 27, 2021·Nature Protocols·Rossella CrescitelliJan Lötvall
Nov 5, 2020·Journal of Extracellular Vesicles·Carolina SoekmadjiLei Zheng
Feb 9, 2021·Frontiers in Cell and Developmental Biology·Giulia MarosticaRoberto Furlan
Nov 19, 2020·International Journal of Molecular Sciences·Leona ChitoiuMarieta Costache
Apr 26, 2020·International Journal of Molecular Sciences·Magdalena SurmanMałgorzata Przybyło
Mar 27, 2021·Analytical Chemistry·Dario BrambillaMarcella Chiari
Aug 21, 2020·Journal for Immunotherapy of Cancer·Jacob J OrmeJeffrey L Winters
Apr 8, 2021·Cell Reports·Tim BeltraminelliMichele De Palma
May 14, 2021·Translational Oncology·Yunjie WuMargaret M Mc Gee
Jul 16, 2021·Journal of Extracellular Vesicles·Kyong-Su ParkJan Lötvall
Sep 15, 2021·Journal of Extracellular Vesicles·Eszter Á TóthEdit I Buzás
Oct 5, 2021·Expert Review of Proteomics·Niraj BabuAditi Chatterjee

❮ Previous
Next ❯

Methods Mentioned

BETA
ELISA
flow cytometry
electron microscopy
Protein Assay
chip
Fusion
chemical fixation
dissection
electrophoresis
transmission electron microscopy

Software Mentioned

TRACK
GraphPad Prism
IMOD
EVpedia
Proteome Discoverer
GraphPad
EV
Percolator
ZetaView®
ExoView

Related Concepts

Related Feeds

Basal Ganglia

Basal Ganglia are a group of subcortical nuclei in the brain associated with control of voluntary motor movements, procedural and habit learning, emotion, and cognition. Here is the latest research.