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

Margaret Simonian; Rachel R.Ogorzalek Loo; Nalaka Rannulu; Joseph A. Loo; Mark P. Molloy; Marcus A. Stoodley

doi:10.1186/s12014-017-9151-3

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

Margaret Simonian^*, Rachel R.Ogorzalek Loo, Nalaka Rannulu, Joseph A. Loo, Mark P. Molloy, Marcus A. Stoodley

^*Corresponding author for this work

Australian Proteome Analysis Facility (APAF)

Research output: Contribution to journal › Article › peer-review

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Abstract

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.

Original language	English
Article number	17
Pages (from-to)	1-12
Number of pages	12
Journal	Clinical Proteomics
Volume	14
DOIs	https://doi.org/10.1186/s12014-017-9151-3
Publication status	Published - 2017

Bibliographical note

Copyright the Author(s) 2017. Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.

Keywords

Endothelial cells
Biotinylation
Membrane proteins
Irradiation
Arteriovenous malformations

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@article{f18f1dcc531b48ef9c462c05c6898d37,

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

abstract = "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 MSE) 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 MSE 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.",

keywords = "Endothelial cells, Biotinylation, Membrane proteins, Irradiation, Arteriovenous malformations",

author = "Margaret Simonian and Loo, {Rachel R.Ogorzalek} and Nalaka Rannulu and Loo, {Joseph A.} and Molloy, {Mark P.} and Stoodley, {Marcus A.}",

note = "Copyright the Author(s) 2017. Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.",

year = "2017",

doi = "10.1186/s12014-017-9151-3",

language = "English",

volume = "14",

pages = "1--12",

journal = "Clinical Proteomics",

issn = "1542-6416",

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TY - JOUR

T1 - Identification of protein targets in cerebral endothelial cells for brain arteriovenous malformation (AVMs) molecular therapies

AU - Simonian, Margaret

AU - Loo, Rachel R.Ogorzalek

AU - Rannulu, Nalaka

AU - Loo, Joseph A.

AU - Molloy, Mark P.

AU - Stoodley, Marcus A.

N1 - Copyright the Author(s) 2017. Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.

PY - 2017

Y1 - 2017

N2 - 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 MSE) 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 MSE 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.

AB - 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 MSE) 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 MSE 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.

KW - Endothelial cells

KW - Biotinylation

KW - Membrane proteins

KW - Irradiation

KW - Arteriovenous malformations

UR - http://www.scopus.com/inward/record.url?scp=85019487962&partnerID=8YFLogxK

U2 - 10.1186/s12014-017-9151-3

DO - 10.1186/s12014-017-9151-3

M3 - Article

C2 - 28522939

AN - SCOPUS:85019487962

SN - 1542-6416

VL - 14

SP - 1

EP - 12

JO - Clinical Proteomics

JF - Clinical Proteomics

M1 - 17

ER -

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

Abstract

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