Dataset Information

CLDN3 in prostate cancer exosomes

ABSTRACT: Exosomes are an emerging source for biomarkers in various diseases. Prostate cancer is the most frequent cancer in men in industrialized countries. The only biomarker widely used is prostate specific antigen with its clinical relevance suffering from low specificity. To identify novel biomarker candidates for prostate cancer we isolated exosomes from the supernatant of malignant PC3 and benign PNT1A cells, grown under standard culture conditions, using sequential centrifugation and ultracentrifugation. Not to alter the cellular growth conditions an FBS-containing medium was used. As control this medium was also subjected to exosomen isolation. Exosome and control samples were analyzed on an LTQ-Orbitrap XL mass spectrometer and subsequently between 441 and 843 proteins per sample were identified by searching against the SwissProt database. 64 proteins exclusively found in PC3 exosomes were subjected to a scoring system, integrating data from five publicly accessible databases and literature search, to identify potential biomarker candidates. Among these the epithelial tight junction protein claudin-3 achieved the highest score. Retests under serum-free conditions using immunoblotting and immunogold-labeling confirmed the expression of claudin-3 on PC3 exosomes. Levels of circulating claudin-3 directly determined from blood plasma, without prior isolation of exosomes, using ELISA were significantly higher (p=0,038) in metastatic prostate cancer patients (n=15) compared to controls with histologically proven benign prostate hyperplasia (n=15). Our results for the first time describe claudin-3 as a potential biomarker candidate in prostate cancer, easily detectably using liquid biopsies, without need for laborious isolation of exosomes. Further studies including larger patient cohorts in different clinical states of prostate cancer are needed to validate our findings.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Homo Sapiens (human) Bos Taurus (bovine)

TISSUE(S): Prostate Adenocarcinoma Cell Line, Prostate Epithelium Cell Line, Epithelial Cell, Blood Serum

DISEASE(S): Prostate Adenocarcinoma

SUBMITTER: Thomas Worst

LAB HEAD: Michael Boutros

PROVIDER: PXD003651 | Pride | 2017-04-13

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	PC3_exo_rep1_high_mol_weight.RAW	Raw
	PC3_exo_rep1_high_mol_weight.RAW.mgf	Mgf
	PC3_exo_rep1_high_mol_weight.dat-pride.pride.mgf.gz	Mgf
	PC3_exo_rep1_high_mol_weight.dat-pride.pride.mztab.gz	Mztab
	PC3_exo_rep1_high_mol_weight.dat-pride.xml.gz	Xml

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Publications

Database-augmented Mass Spectrometry Analysis of Exosomes Identifies Claudin 3 as a Putative Prostate Cancer Biomarker.

Worst Thomas Stefan TS von Hardenberg Jost J Gross Julia Christina JC Erben Philipp P Schnölzer Martina M Hausser Ingrid I Bugert Peter P Michel Maurice Stephan MS Boutros Michael M

Molecular & cellular proteomics : MCP 20170409 6

In prostate cancer and other malignancies sensitive and robust biomarkers are lacking or have relevant limitations. Prostate specific antigen (PSA), the only biomarker widely used in prostate cancer, is suffering from low specificity. Exosomes offer new perspectives in the discovery of blood-based biomarkers. Here we present a proof-of principle study for a proteomics-based identification pipeline, implementing existing data sources, to exemplarily identify exosome-based biomarker candidates in ...[more]

PMID: 28396511

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