Proteomics analysis of serum small extracellular vesicles for the longitudinal study of a glioblastoma multiforme mouse model

Abstract Longitudinal analysis of disease models enables the molecular changes due to disease progression or therapeutic intervention to be better resolved. Approximately 75 µl of serum can be drawn from a mouse every 14 days. To date no methods have been reported that are able to analyze the proteo...

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Autores principales:	Federica Anastasi, Francesco Greco, Marialaura Dilillo, Eleonora Vannini, Valentina Cappello, Laura Baroncelli, Mario Costa, Mauro Gemmi, Matteo Caleo, Liam A. McDonnell
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/ce1a6757329b4d20a2270c703db00157
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spelling	oai:doaj.org-article:ce1a6757329b4d20a2270c703db001572021-12-02T11:40:49ZProteomics analysis of serum small extracellular vesicles for the longitudinal study of a glioblastoma multiforme mouse model10.1038/s41598-020-77535-82045-2322https://doaj.org/article/ce1a6757329b4d20a2270c703db001572020-11-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-77535-8https://doaj.org/toc/2045-2322Abstract Longitudinal analysis of disease models enables the molecular changes due to disease progression or therapeutic intervention to be better resolved. Approximately 75 µl of serum can be drawn from a mouse every 14 days. To date no methods have been reported that are able to analyze the proteome of small extracellular vesicles (sEV’s) from such low serum volumes. Here we report a method for the proteomics analysis of sEV's from 50 µl of serum. Two sEV isolation procedures were first compared; precipitation based purification (PPT) and size exclusion chromatography (SEC). The methodological comparison confirmed that SEC led to purer sEV’s both in terms of size and identified proteins. The procedure was then scaled down and the proteolytic digestion further optimized. The method was then applied to a longitudinal study of serum-sEV proteome changes in a glioblastoma multiforme (GBM) mouse model. Serum was collected at multiple time points, sEV’s isolated and their proteins analyzed. The protocol enabled 274 protein groups to be identified and quantified. The longitudinal analysis revealed 25 deregulated proteins in GBM serum sEV's including proteins previously shown to be associated with GBM progression and metastasis (Myh9, Tln-1, Angpt1, Thbs1).Federica AnastasiFrancesco GrecoMarialaura DililloEleonora VanniniValentina CappelloLaura BaroncelliMario CostaMauro GemmiMatteo CaleoLiam A. McDonnellNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-11 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Federica Anastasi Francesco Greco Marialaura Dilillo Eleonora Vannini Valentina Cappello Laura Baroncelli Mario Costa Mauro Gemmi Matteo Caleo Liam A. McDonnell Proteomics analysis of serum small extracellular vesicles for the longitudinal study of a glioblastoma multiforme mouse model
description	Abstract Longitudinal analysis of disease models enables the molecular changes due to disease progression or therapeutic intervention to be better resolved. Approximately 75 µl of serum can be drawn from a mouse every 14 days. To date no methods have been reported that are able to analyze the proteome of small extracellular vesicles (sEV’s) from such low serum volumes. Here we report a method for the proteomics analysis of sEV's from 50 µl of serum. Two sEV isolation procedures were first compared; precipitation based purification (PPT) and size exclusion chromatography (SEC). The methodological comparison confirmed that SEC led to purer sEV’s both in terms of size and identified proteins. The procedure was then scaled down and the proteolytic digestion further optimized. The method was then applied to a longitudinal study of serum-sEV proteome changes in a glioblastoma multiforme (GBM) mouse model. Serum was collected at multiple time points, sEV’s isolated and their proteins analyzed. The protocol enabled 274 protein groups to be identified and quantified. The longitudinal analysis revealed 25 deregulated proteins in GBM serum sEV's including proteins previously shown to be associated with GBM progression and metastasis (Myh9, Tln-1, Angpt1, Thbs1).
format	article
author	Federica Anastasi Francesco Greco Marialaura Dilillo Eleonora Vannini Valentina Cappello Laura Baroncelli Mario Costa Mauro Gemmi Matteo Caleo Liam A. McDonnell
author_facet	Federica Anastasi Francesco Greco Marialaura Dilillo Eleonora Vannini Valentina Cappello Laura Baroncelli Mario Costa Mauro Gemmi Matteo Caleo Liam A. McDonnell
author_sort	Federica Anastasi
title	Proteomics analysis of serum small extracellular vesicles for the longitudinal study of a glioblastoma multiforme mouse model
title_short	Proteomics analysis of serum small extracellular vesicles for the longitudinal study of a glioblastoma multiforme mouse model
title_full	Proteomics analysis of serum small extracellular vesicles for the longitudinal study of a glioblastoma multiforme mouse model
title_fullStr	Proteomics analysis of serum small extracellular vesicles for the longitudinal study of a glioblastoma multiforme mouse model
title_full_unstemmed	Proteomics analysis of serum small extracellular vesicles for the longitudinal study of a glioblastoma multiforme mouse model
title_sort	proteomics analysis of serum small extracellular vesicles for the longitudinal study of a glioblastoma multiforme mouse model
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/ce1a6757329b4d20a2270c703db00157
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Proteomics analysis of serum small extracellular vesicles for the longitudinal study of a glioblastoma multiforme mouse model

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