Large and small extracellular vesicles released by glioma cells in vitro and in vivo
Tumour cells release diverse populations of extracellular vesicles (EVs) ranging in size, molecular cargo, and function. We sought to characterize mRNA and protein content of EV subpopulations released by human glioblastoma (GBM) cells expressing a mutant form of epidermal growth factor receptor (U8...
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doaj-8cd2c4798cf54a19a0cbbe15d8d6a0ba2020-11-25T03:37:07ZengTaylor & Francis GroupJournal of Extracellular Vesicles2001-30782020-01-019110.1080/20013078.2019.16897841689784Large and small extracellular vesicles released by glioma cells in vitro and in vivoAnudeep Yekula0Valentina R. Minciacchi1Matteo Morello2Huilin Shao3Yongil Park4Xuan Zhang5Koushik Muralidharan6Michael R. Freeman7Ralph Weissleder8Hakho Lee9Bob Carter10Xandra O. Breakefield11Dolores Di Vizio12Leonora Balaj13Massachusetts General HospitalPathology & Laboratory Medicine, Samuel Oschin Comprehensive Cancer Institute, Cedars-Sinai Medical CenterPathology & Laboratory Medicine, Samuel Oschin Comprehensive Cancer Institute, Cedars-Sinai Medical CenterMassachusetts General HospitalMassachusetts General HospitalMassachusetts General Hospital and Harvard Medical SchoolMassachusetts General HospitalPathology & Laboratory Medicine, Samuel Oschin Comprehensive Cancer Institute, Cedars-Sinai Medical CenterMassachusetts General HospitalMassachusetts General HospitalMassachusetts General HospitalMassachusetts General Hospital and Harvard Medical SchoolPathology & Laboratory Medicine, Samuel Oschin Comprehensive Cancer Institute, Cedars-Sinai Medical CenterMassachusetts General HospitalTumour cells release diverse populations of extracellular vesicles (EVs) ranging in size, molecular cargo, and function. We sought to characterize mRNA and protein content of EV subpopulations released by human glioblastoma (GBM) cells expressing a mutant form of epidermal growth factor receptor (U87EGFRvIII) in vitro and in vivo with respect to size, morphology and the presence of tumour cargo. The two EV subpopulations purified from GBM U87EGFRvIII cancer cells, non-cancer human umbilical vein endothelial cells (HUVEC; control) and serum of U87EGFRvIII glioma-bearing mice using differential centrifugation (EVs that sediment at 10,000 × g or 100,000 × g are termed large EVs and small EVs, respectively) were characterized using transmission electron microscopy (TEM), confocal microscopy, nanoparticle tracking analysis (NTA), flow cytometry, immunofluorescence (IF), quantitative-polymerase chain reaction (qPCR), droplet digital polymerase chain reaction (ddPCR) and micro-nuclear magnetic resonance (μNMR). We report that both U87EGFRvIII and HUVEC release a similar number of small EVs, but U87EGFRvIII glioma cells alone release a higher number of large EVs compared to non-cancer HUVEC. The EGFRvIII mRNA from the two EV subpopulations from U87EGFRvIII glioma cells was comparable, while the EGFR protein (wild type + vIII) levels are significantly higher in large EVs. Similarly, EGFRvIII mRNA in large and small EVs isolated from the serum of U87EGFRvIII glioma-bearing mice is comparable, while the EGFR protein (wild type + vIII) levels are significantly higher in large EVs. Here we report for the first time a direct comparison of large and small EVs released by glioma U87EGFRvIII cells and from serum of U87EGFRvIII glioma-bearing mice. Both large and small EVs contain tumour-specific EGFRvIII mRNA and proteins and combining these platforms may be beneficial in detecting rare mutant events in circulating biofluids.http://dx.doi.org/10.1080/20013078.2019.1689784extracellular vesicleslarge extracellular vesiclessmall extracellular vesiclesbiomarkersglioma |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Anudeep Yekula Valentina R. Minciacchi Matteo Morello Huilin Shao Yongil Park Xuan Zhang Koushik Muralidharan Michael R. Freeman Ralph Weissleder Hakho Lee Bob Carter Xandra O. Breakefield Dolores Di Vizio Leonora Balaj |
spellingShingle |
Anudeep Yekula Valentina R. Minciacchi Matteo Morello Huilin Shao Yongil Park Xuan Zhang Koushik Muralidharan Michael R. Freeman Ralph Weissleder Hakho Lee Bob Carter Xandra O. Breakefield Dolores Di Vizio Leonora Balaj Large and small extracellular vesicles released by glioma cells in vitro and in vivo Journal of Extracellular Vesicles extracellular vesicles large extracellular vesicles small extracellular vesicles biomarkers glioma |
author_facet |
Anudeep Yekula Valentina R. Minciacchi Matteo Morello Huilin Shao Yongil Park Xuan Zhang Koushik Muralidharan Michael R. Freeman Ralph Weissleder Hakho Lee Bob Carter Xandra O. Breakefield Dolores Di Vizio Leonora Balaj |
author_sort |
Anudeep Yekula |
title |
Large and small extracellular vesicles released by glioma cells in vitro and in vivo |
title_short |
Large and small extracellular vesicles released by glioma cells in vitro and in vivo |
title_full |
Large and small extracellular vesicles released by glioma cells in vitro and in vivo |
title_fullStr |
Large and small extracellular vesicles released by glioma cells in vitro and in vivo |
title_full_unstemmed |
Large and small extracellular vesicles released by glioma cells in vitro and in vivo |
title_sort |
large and small extracellular vesicles released by glioma cells in vitro and in vivo |
publisher |
Taylor & Francis Group |
series |
Journal of Extracellular Vesicles |
issn |
2001-3078 |
publishDate |
2020-01-01 |
description |
Tumour cells release diverse populations of extracellular vesicles (EVs) ranging in size, molecular cargo, and function. We sought to characterize mRNA and protein content of EV subpopulations released by human glioblastoma (GBM) cells expressing a mutant form of epidermal growth factor receptor (U87EGFRvIII) in vitro and in vivo with respect to size, morphology and the presence of tumour cargo. The two EV subpopulations purified from GBM U87EGFRvIII cancer cells, non-cancer human umbilical vein endothelial cells (HUVEC; control) and serum of U87EGFRvIII glioma-bearing mice using differential centrifugation (EVs that sediment at 10,000 × g or 100,000 × g are termed large EVs and small EVs, respectively) were characterized using transmission electron microscopy (TEM), confocal microscopy, nanoparticle tracking analysis (NTA), flow cytometry, immunofluorescence (IF), quantitative-polymerase chain reaction (qPCR), droplet digital polymerase chain reaction (ddPCR) and micro-nuclear magnetic resonance (μNMR). We report that both U87EGFRvIII and HUVEC release a similar number of small EVs, but U87EGFRvIII glioma cells alone release a higher number of large EVs compared to non-cancer HUVEC. The EGFRvIII mRNA from the two EV subpopulations from U87EGFRvIII glioma cells was comparable, while the EGFR protein (wild type + vIII) levels are significantly higher in large EVs. Similarly, EGFRvIII mRNA in large and small EVs isolated from the serum of U87EGFRvIII glioma-bearing mice is comparable, while the EGFR protein (wild type + vIII) levels are significantly higher in large EVs. Here we report for the first time a direct comparison of large and small EVs released by glioma U87EGFRvIII cells and from serum of U87EGFRvIII glioma-bearing mice. Both large and small EVs contain tumour-specific EGFRvIII mRNA and proteins and combining these platforms may be beneficial in detecting rare mutant events in circulating biofluids. |
topic |
extracellular vesicles large extracellular vesicles small extracellular vesicles biomarkers glioma |
url |
http://dx.doi.org/10.1080/20013078.2019.1689784 |
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