Preprint Article Version 1 NOT YET PEER-REVIEWED

Characterization of Extracellular Vesicles Isolated from Plasma and Urine of Healthy Donors and
Cancer Patients

  1. Experimental Oncology, Ethics and Palliative Care in Oncology, University Hospital and Medical School, UKSH, Luebeck 23562, Germany
  2. Clinic for Rheumatology, University Hospital and Medical School, UKSH, Luebeck 23562, Germany
  3. Experimental Nephrology, University Hospital and Medical School, UKSH, Luebeck 23562, Germany
Version 1 : Received: 13 October 2016 / Approved: 14 October 2016 / Online: 14 October 2016 (11:09:13 CEST)

How to cite: Freund, A.; Plattfaut, C.; Mueller, A.; Riemekasten, G.; Haas, C.; Gieseler, F. Characterization of Extracellular Vesicles Isolated from Plasma and Urine of Healthy Donors and Cancer Patients. Preprints 2016, 2016100056 (doi: 10.20944/preprints201610.0056.v1). Freund, A.; Plattfaut, C.; Mueller, A.; Riemekasten, G.; Haas, C.; Gieseler, F. Characterization of Extracellular Vesicles Isolated from Plasma and Urine of Healthy Donors and Cancer Patients. Preprints 2016, 2016100056 (doi: 10.20944/preprints201610.0056.v1).

Abstract

Extracellular vesicles (EVs) were detected and quantified in blood and urine of healthy donors and cancer patients using serial centrifugation protocols and high- resolution flow cytometry. EVs were phenotypically characterized by measuring exposure of phosphatidylserine (PS) and activities of FX and TFPI as well as functionally analyzed for tenase activity, ERK phosphorylation and tumor cell migration. Plasmatic and urinary EVs contained mainly microparticles (MPs), accompanied by low amounts of exosomes or apoptotic bodies. In contrast to plasmatic MPs, urinary MPs from healthy donors did not expose PS, which argues for endothelial cells of the urinary tract as a source. In cancer patients, however, we found urinary MPs presenting PS. Their amount correlated positively with albumin excretion. This suggests that the glomerular membrane is leaky for plasmatic MPs as well as albumin; thus in the urine of cancer patients the EVs presenting PS might be obtained from the plasma. Positive correlations between tenase activity, ERK phosphorylation and potency to induce tumor cell migration suggest that TF-presenting EVs form PAR- activating tenase complexes with FVII and FX, activate PAR-2, induce ERK phosphorylation and trigger tumor cell migration. Our results representing proof-of-principle suggest that further studies in defined cancers and stages are warranted.

Subject Areas

extracellular vesicles; microparticles; protease-activated receptors; tumor cell migration

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