Dataset Information

Recombinant extracellular vesicles as a biological reference material

ABSTRACT: Recent years have seen a tremendous increase in the study of extracellular vesicles (EV) geared towards biological understanding, diagnostics and therapy. The interpretation of EV data however remains challenging owing to the complexity of biofluids and the technical variability that are introduced during sample preparation. To understand and mitigate these errors we present the use of traceable recombinant EV (rEV) that mimic endogenous EV both physically and biochemically. The informed use of rEV ensures standardized measurements in various research and biomedical applications. In this dataset we performed mass spectrometry-based proteomics analyses to compare the protein content of endogenous EVs (eEV) and traceable recombinant EV (rEV). The analysis showed that both types of EVs are very similar and that more than 90% of proteins are equally abundant in rEV and eEV isolated from HEK293T cells.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Hek-293t Cell, Kidney Cell

SUBMITTER: Evy Timmerman

LAB HEAD: An Hendrix

PROVIDER: PXD010269 | Pride | 2019-07-11

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	E13394b_1p_6248_Dewever_EV_Mock_1.raw	Raw
	E13396b_1p_6248_Dewever_EV_Mock_2.raw	Raw
	E13398b_1p_6248_Dewever_EV_Mock_3.raw	Raw
	E13400b_1p_6248_Dewever_VLP_1.raw	Raw
	E13402b_1p_6248_Dewever_VLP_2.raw	Raw

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Publications

The generation and use of recombinant extracellular vesicles as biological reference material.

Geeurickx Edward E Tulkens Joeri J Dhondt Bert B Van Deun Jan J Lippens Lien L Vergauwen Glenn G Heyrman Elisa E De Sutter Delphine D Gevaert Kris K Impens Francis F Miinalainen Ilkka I Van Bockstal Pieter-Jan PJ De Beer Thomas T Wauben Marca H M MHM Nolte-'t-Hoen Esther N M ENM Bloch Katarzyna K Swinnen Johannes V JV van der Pol Edwin E Nieuwland Rienk R Braems Geert G Callewaert Nico N Mestdagh Pieter P Vandesompele Jo J Denys Hannelore H Eyckerman Sven S De Wever Olivier O Hendrix An A

Nature communications 20190723 1

Recent years have seen an increase of extracellular vesicle (EV) research geared towards biological understanding, diagnostics and therapy. However, EV data interpretation remains challenging owing to complexity of biofluids and technical variation introduced during sample preparation and analysis. To understand and mitigate these limitations, we generated trackable recombinant EV (rEV) as a biological reference material. Employing complementary characterization methods, we demonstrate that rEV ...[more]

PMID: 31337761

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Project description:Background & Aims: Extracellular vesicles (EVs) play a pivotal role in connecting tumor cells and their local and distant microenvironment. Here, we aimed to understand the role and molecular basis of patient-derived EVs in modulating cancer stemness and tumorigenesis in the context of hepatocellular carcinoma (HCC). Methods: EVs were isolated, quantified and characterized from patients’ sera. EVs were tested vigorously, both in vitro and in vivo, by various functional assays. Proteomic analysis was performed to identify the functional components of EVs. The expression level of polymeric immunoglobulin receptor (pIgR) in circulating EVs, tumor and non-tumorous tissues of HCC patients was determined by ELISA, immunoblotting, immunohistochemistry and quantitative PCR. The functional role and underlying mechanism of EVs with an enhanced pIgR expression was elucidated. Blockage of EV-pIgR with neutralizing antibody was performed in nude mice implanted with patient-derived tumor xenograft (PDTX). Results: Circulating EVs of late-stage HCC (L-HCC) patient had significantly elevated pIgR expression when compared to the EVs released by control individuals. The augmenting effect of L-HCC patient in cancer stemness and tumorigenesis was hindered by anti-pIgR antibody. EVs enriched with pIgR consistently promoted cancer stemness and cancerous phenotypes in the recipient cells. Mechanistically, EV-pIgR-induced cancer aggressiveness was abrogated by Akt and β-catenin inhibitors, ascertaining the role of EV-pIgR through the activation of PDK1/Akt/GSK3β/β-catenin signaling axis. Furthermore, anti-pIgR neutralizing antibody attenuated the tumor growth in mice implanted with PDTX. Conclusion: The study illustrates an unrevealed role of EV-pIgR in regulating cancer stemness and aggressiveness, in which EV-pIgR activates PDK1/Akt/GSK3β/β-catenin signaling cascades. The blockage of intercellular communications mediated by EV-pIgR in the tumor microenvironment may provide a new therapeutic strategy for cancer patients.