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Global extracellular vesicle proteomic signature defines U87-MG glioma cell hypoxic status with potential implications for non-invasive diagnostics

Indira Chandran, Vineesh LU ; Welinder, Charlotte LU ; Gonçalves de Oliveira, Kelin LU ; Cerezo-Magaña, Myriam LU ; Månsson, Ann Sofie LU ; Johansson, Maria C. LU ; Marko-Varga, Gyorgy LU and Belting, Mattias LU (2019) In Journal of Neuro-Oncology 144(3). p.477-488
Abstract

Purpose: Glioblastoma multiforme (GBM) is the most common and lethal of primary malignant brain tumors. Hypoxia constitutes a major determining factor for the poor prognosis of high-grade glioma patients, and is known to contribute to the development of treatment resistance. Therefore, new strategies to comprehensively profile and monitor the hypoxic status of gliomas are of high clinical relevance. Here, we have explored how the proteome of secreted extracellular vesicles (EVs) at the global level may reflect hypoxic glioma cells. Methods: We have employed shotgun proteomics and label free quantification to profile EVs isolated from human high-grade glioma U87-MG cells cultured at normoxia or hypoxia. Parallel reaction monitoring was... (More)

Purpose: Glioblastoma multiforme (GBM) is the most common and lethal of primary malignant brain tumors. Hypoxia constitutes a major determining factor for the poor prognosis of high-grade glioma patients, and is known to contribute to the development of treatment resistance. Therefore, new strategies to comprehensively profile and monitor the hypoxic status of gliomas are of high clinical relevance. Here, we have explored how the proteome of secreted extracellular vesicles (EVs) at the global level may reflect hypoxic glioma cells. Methods: We have employed shotgun proteomics and label free quantification to profile EVs isolated from human high-grade glioma U87-MG cells cultured at normoxia or hypoxia. Parallel reaction monitoring was used to quantify the identified, hypoxia-associated EV proteins. To determine the potential biological significance of hypoxia-associated proteins, the cumulative Z score of identified EV proteins was compared with GBM subtypes from HGCC and TCGA databases. Results: In total, 2928 proteins were identified in EVs, out of which 1654 proteins overlapped with the ExoCarta EV-specific database. We found 1034 proteins in EVs that were unique to the hypoxic status of U87-MG cells. We subsequently identified an EV protein signature, “HYPSIGNATURE”, encompassing nine proteins that strongly represented the hypoxic situation and exhibited close proximity to the mesenchymal GBM subtype. Conclusions: We propose, for the first time, an EV protein signature that could comprehensively reflect the hypoxic status of high-grade glioma cells. The presented data provide proof-of-concept for targeted proteomic profiling of glioma derived EVs, which should motivate future studies exploring its utility in non-invasive diagnosis and monitoring of brain tumor patients.

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author
; ; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Extracellular vesicles, Glioblastoma, Hypoxia, Label free quantification, Mass spectrometry
in
Journal of Neuro-Oncology
volume
144
issue
3
pages
477 - 488
publisher
Springer
external identifiers
  • pmid:31414377
  • scopus:85070778611
ISSN
0167-594X
DOI
10.1007/s11060-019-03262-4
language
English
LU publication?
yes
id
ca3b0c58-0729-4b3c-a11b-a0c95b3cbb15
date added to LUP
2019-09-04 12:54:15
date last changed
2021-06-23 02:56:48
@article{ca3b0c58-0729-4b3c-a11b-a0c95b3cbb15,
  abstract     = {<p>Purpose: Glioblastoma multiforme (GBM) is the most common and lethal of primary malignant brain tumors. Hypoxia constitutes a major determining factor for the poor prognosis of high-grade glioma patients, and is known to contribute to the development of treatment resistance. Therefore, new strategies to comprehensively profile and monitor the hypoxic status of gliomas are of high clinical relevance. Here, we have explored how the proteome of secreted extracellular vesicles (EVs) at the global level may reflect hypoxic glioma cells. Methods: We have employed shotgun proteomics and label free quantification to profile EVs isolated from human high-grade glioma U87-MG cells cultured at normoxia or hypoxia. Parallel reaction monitoring was used to quantify the identified, hypoxia-associated EV proteins. To determine the potential biological significance of hypoxia-associated proteins, the cumulative Z score of identified EV proteins was compared with GBM subtypes from HGCC and TCGA databases. Results: In total, 2928 proteins were identified in EVs, out of which 1654 proteins overlapped with the ExoCarta EV-specific database. We found 1034 proteins in EVs that were unique to the hypoxic status of U87-MG cells. We subsequently identified an EV protein signature, “HYP<sub>SIGNATURE</sub>”, encompassing nine proteins that strongly represented the hypoxic situation and exhibited close proximity to the mesenchymal GBM subtype. Conclusions: We propose, for the first time, an EV protein signature that could comprehensively reflect the hypoxic status of high-grade glioma cells. The presented data provide proof-of-concept for targeted proteomic profiling of glioma derived EVs, which should motivate future studies exploring its utility in non-invasive diagnosis and monitoring of brain tumor patients.</p>},
  author       = {Indira Chandran, Vineesh and Welinder, Charlotte and Gonçalves de Oliveira, Kelin and Cerezo-Magaña, Myriam and Månsson, Ann Sofie and Johansson, Maria C. and Marko-Varga, Gyorgy and Belting, Mattias},
  issn         = {0167-594X},
  language     = {eng},
  month        = {08},
  number       = {3},
  pages        = {477--488},
  publisher    = {Springer},
  series       = {Journal of Neuro-Oncology},
  title        = {Global extracellular vesicle proteomic signature defines U87-MG glioma cell hypoxic status with potential implications for non-invasive diagnostics},
  url          = {http://dx.doi.org/10.1007/s11060-019-03262-4},
  doi          = {10.1007/s11060-019-03262-4},
  volume       = {144},
  year         = {2019},
}