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Identification of protein targets in cerebral endothelial cells for brain arteriovenous malformation (AVMs) molecular therapies

BACKGROUND: To develop a new molecular targeted treatment for brain (AVMs), identification of membrane proteins that are localised on the AVM endothelium is crucial. Current treatment methods are surgery and radiosurgery. However, complete occlusion post radiosurgery are achieved within 3 years, whi...

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Autores principales: Simonian, Margaret, Loo, Rachel R. Ogorzalek, Rannulu, Nalaka, Loo, Joseph A., Molloy, Mark P., Stoodley, Marcus A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5434588/
https://www.ncbi.nlm.nih.gov/pubmed/28522939
http://dx.doi.org/10.1186/s12014-017-9151-3
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author Simonian, Margaret
Loo, Rachel R. Ogorzalek
Rannulu, Nalaka
Loo, Joseph A.
Molloy, Mark P.
Stoodley, Marcus A.
author_facet Simonian, Margaret
Loo, Rachel R. Ogorzalek
Rannulu, Nalaka
Loo, Joseph A.
Molloy, Mark P.
Stoodley, Marcus A.
author_sort Simonian, Margaret
collection PubMed
description BACKGROUND: To develop a new molecular targeted treatment for brain (AVMs), identification of membrane proteins that are localised on the AVM endothelium is crucial. Current treatment methods are surgery and radiosurgery. However, complete occlusion post radiosurgery are achieved within 3 years, while patient remain at risk of haemorrhage. This study aims to identify potential protein targets in AVM endothelial cells that discriminate these vessels from normal vessels; these proteins targets will be investigated for the molecular therapy of brain AVMs to promote rapid thrombosis after radiosurgery. METHODS: We employed in vitro biotinylation that we developed, and mass spectrometry to detect cell surface-exposed proteins in cultures of murine cerebral endothelial cells (bEnd.3). Two forms of mass spectrometry were applied (iTRAQ-MS and MS(E)) to identify and quantify membrane protein expression at various time-points following irradiation which simulates a radiosurgical treatment approach. Immunocytochemistry was used to confirm the expression of selected membrane proteins. ProteinPilot V4.0 software was used to analyse the iTRAQ-MS data and the MS(E) data was analysed using ProteinLynx Global Server version 2.5 software. RESULTS: The proteomics data revealed several differentially expressed membrane proteins between irradiated and non-irradiated cells at specific time points, e.g. PECAM-1, cadherin-5, PDI, EPCR and integrins. Immunocytochemistry data confirmed the expression of these proteins. CONCLUSION: Cell surface protein biotinylation and proteomics analysis successfully identified membrane proteins from murine brain endothelial cells in response to irradiation. This work suggests potential target protein molecules for evaluation in animal models of brain-AVM. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12014-017-9151-3) contains supplementary material, which is available to authorized users.
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spelling pubmed-54345882017-05-18 Identification of protein targets in cerebral endothelial cells for brain arteriovenous malformation (AVMs) molecular therapies Simonian, Margaret Loo, Rachel R. Ogorzalek Rannulu, Nalaka Loo, Joseph A. Molloy, Mark P. Stoodley, Marcus A. Clin Proteomics Research BACKGROUND: To develop a new molecular targeted treatment for brain (AVMs), identification of membrane proteins that are localised on the AVM endothelium is crucial. Current treatment methods are surgery and radiosurgery. However, complete occlusion post radiosurgery are achieved within 3 years, while patient remain at risk of haemorrhage. This study aims to identify potential protein targets in AVM endothelial cells that discriminate these vessels from normal vessels; these proteins targets will be investigated for the molecular therapy of brain AVMs to promote rapid thrombosis after radiosurgery. METHODS: We employed in vitro biotinylation that we developed, and mass spectrometry to detect cell surface-exposed proteins in cultures of murine cerebral endothelial cells (bEnd.3). Two forms of mass spectrometry were applied (iTRAQ-MS and MS(E)) to identify and quantify membrane protein expression at various time-points following irradiation which simulates a radiosurgical treatment approach. Immunocytochemistry was used to confirm the expression of selected membrane proteins. ProteinPilot V4.0 software was used to analyse the iTRAQ-MS data and the MS(E) data was analysed using ProteinLynx Global Server version 2.5 software. RESULTS: The proteomics data revealed several differentially expressed membrane proteins between irradiated and non-irradiated cells at specific time points, e.g. PECAM-1, cadherin-5, PDI, EPCR and integrins. Immunocytochemistry data confirmed the expression of these proteins. CONCLUSION: Cell surface protein biotinylation and proteomics analysis successfully identified membrane proteins from murine brain endothelial cells in response to irradiation. This work suggests potential target protein molecules for evaluation in animal models of brain-AVM. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12014-017-9151-3) contains supplementary material, which is available to authorized users. BioMed Central 2017-05-16 /pmc/articles/PMC5434588/ /pubmed/28522939 http://dx.doi.org/10.1186/s12014-017-9151-3 Text en © The Author(s) 2017 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Simonian, Margaret
Loo, Rachel R. Ogorzalek
Rannulu, Nalaka
Loo, Joseph A.
Molloy, Mark P.
Stoodley, Marcus A.
Identification of protein targets in cerebral endothelial cells for brain arteriovenous malformation (AVMs) molecular therapies
title Identification of protein targets in cerebral endothelial cells for brain arteriovenous malformation (AVMs) molecular therapies
title_full Identification of protein targets in cerebral endothelial cells for brain arteriovenous malformation (AVMs) molecular therapies
title_fullStr Identification of protein targets in cerebral endothelial cells for brain arteriovenous malformation (AVMs) molecular therapies
title_full_unstemmed Identification of protein targets in cerebral endothelial cells for brain arteriovenous malformation (AVMs) molecular therapies
title_short Identification of protein targets in cerebral endothelial cells for brain arteriovenous malformation (AVMs) molecular therapies
title_sort identification of protein targets in cerebral endothelial cells for brain arteriovenous malformation (avms) molecular therapies
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5434588/
https://www.ncbi.nlm.nih.gov/pubmed/28522939
http://dx.doi.org/10.1186/s12014-017-9151-3
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