Dataset Information

Proteomic analysis of serum small extracellular vesicles of individual mice for longitudinal investigations

ABSTRACT: Characterization of small extracellular vesicles (sEV) from individual mice for the longitudinal investigation of proteome changes in a mouse model of Glioblastoma Multiforme (GBM). We first compared precipitation and size exclusion chromatography (SEC) for sEV purification from 100ul of mouse serum. We then optimized SEC-based procedure for the isolation of sEV’s from 50ul of mouse serum. The optimized procedure was then applied to compare the sEV proteomes for the longitudinal analysis of the GBM-mouse model.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Blood Serum

DISEASE(S): Glioblastoma

SUBMITTER: Federica Anastasi

LAB HEAD: Liam A. McDonnell

PROVIDER: PXD016473 | Pride | 2020-11-30

REPOSITORIES: Pride

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Publications

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

Anastasi Federica F Greco Francesco F Dilillo Marialaura M Vannini Eleonora E Cappello Valentina V Baroncelli Laura L Costa Mario M Gemmi Mauro M Caleo Matteo M McDonnell Liam A LA

Scientific reports 20201124 1

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; preci ...[more]

PMID: 33235327

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Project description:In patients with severe kidney disease AVF are surgically placed in order to create good access for hemodialysis. In these dialysis patients the failure rates of AVF can be as high as 24% within 6 months after surgery, causing ineffective dialysis and necessitating additional clinical interventions. The pathological processes known to lead to AVF failure are beginning to be unravelled include the formation of venous neointimal hyperplasia (VNIH), thrombosis (Chang et al. PMID 16105066), and venous stenosis (Kanterman et al. PMID7892454, Tang et al. 1998), resulting in a reduced blood flow through the fistula. We established a rat model for AVF failure in human kidney dialysis patients. The characterization of this model has been previously described (Globerman et al. 2011, PubmedID:22002501). In this model the AVFs are surgically constructed in the right leg by connecting the superficial epigastric vein SEV to the common femoral artery (CFA), resulting in exposure of the SEV to arterial pressure with pulsatile and low resistant flow patterns (Globerman et al. 2011, PMID22002501). In the present study we utilized this AVF model in order to assess the effects of arterialized flow, with consequent pathological changes of the vessel wall due to surgical AVF instalment, on the transcriptome of endothelium from the SEV. Within the SEV these pathologies of the vessel wall include the formation of NIH in the main branch, and stenosis in the side branches. By employing this rat model we assessed the changes of the endothelial transcriptome in relation to these pathologies in order to gain mechanistic understanding of the potential roles of venous endothelium in AVF failure, as well as to identify potential biomarkers preceding AVF failure. AVF surgery was performed on N=6 rats, and 13-14 days after surgery the rats were sacrificed. AVFs were extracted from the rats, and microarray transcriptome analyses were performed on luminal endothelial cells that isolated from four different sites of the AVF by employing Laser Capture Microdissection (LCM). These sites included (i) N=5 AVF sections of the superficial epigastric vein (SEV) with neointimal hyperplasia (NIH), (ii) N=3 AVF sections of SEV side branches with luminal stenosis, (iii), N=6 sections of the SEV located distally from a ligation thereby omitting exposure to arterialized flow and consequently preventing the development of NIH or stenosis, and (iv) N=3 sections of the AVF common femoral artery without signs of NIH or stenosis.

Dataset Information

Proteomic analysis of serum small extracellular vesicles of individual mice for longitudinal investigations

Publications

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

Similar Datasets

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