PhD Scientific Days 2020

Molecular Sciences III. Lectures

Surface Protein Interactome of Extracellular Vesicles in Blood Plasma

Előadó neve

Dr. Tóth, Eszter Ágnes

Előadó munkahelye

Department of Genetics, Cell and Immunobiology

Előadó telefonszáma

+36206701135

Előadó e-mail címe

esztr.toth@gmail.com

Az előadás címe

Surface Protein Interactome of Extracellular Vesicles in Blood Plasma

Szerző(k) neve és munkahelye

Eszter Á Tóth; Semmelweis University, Department of Genetics, Cell- and Immunobiology; Budapest
Katalin É Szabó-Taylor; Semmelweis University, Department of Genetics, Cell- and Immunobiology; Budapest
Edit I Buzás; Semmelweis University, Department of Genetics, Cell- and Immunobiology; Budapest

Szekció

Molecular Sciences III. Lectures

Language of the presentation

English

Section, first choice

Molecular Sciences

Section, second choice

Health Sciences

Összefoglaló szövege

Introduction, aims
Extracellular vesicles (EVs) are the endogenous nanoparticles produced by the cells. Artificial nanoparticles have been found to develop a protein corona altering their biodistribution and bioavailability in biological media. Here we set the aim to study if a similar protein corona is formed at the surface of EVs in biofluids and if inflammation had an effect on the protein corona formation. Also, we tested, whether the protein corona alters the biological function of EVs.

Methods
Blood plasma depleted in both platelets and EVs was generated from blood samples of healthy subjects (n=18) and rheumatoid arthritis patients (n=16). Nascent EVs of THP1 cells and platelets were isolated and incubated in different plasma samples for 30 minutes. EVs were then re-isolated by differential centrifugation, size exclusion choramatography (SEC), or density gradient ultracentrifugation (DGUC), and were studied by mass spectrometry (MS/MS), electron microscopy (EM) and flow cytometry. Controls included i) plasma without the addition of EVs and ii) EVs incubated in buffer. The effect of the protein corona was studied by exposing dendritic cells to EVs, after which flow cytometric analysis of activation markers and ELISA of the supernatant were performed.

Results
After subtracting the proteins found by MS/MS in pure (nascent) EV samples from the list of proteins of EV samples incubated in plasma for 30 minutes (coated EVs), a high number of proteins were found, out of which several were more characteristic of rheumatoid arthritis than of healthy samples. DGUC combined with a flow cytometry revealed a significantly higher density of coated vesicles compared to the nascent ones. Interactions between several blood plasma proteins and EVs were also confirmed by flow cytometry and immune EM. Functional assays showed that coated vesicles induce a more pronounced activation response (eg. surface CD83, TNFα production) of dendritic cells.

Conclusion
Our data suggest the formation of a protein corona on EVs of blood plasma. The differences in protein coronas formed in healthy and rheumatoid arthritis plasma samples, suggest that EV surface-associated proteins may play a role in disease pathology and may serve as biomarkers.

Funding
NKFIH-OTKA 111958; ÚNKP-19-3-I-SE-45 – New National Excellence Program of the Ministry for Innovation and Technology

Additional Information

Supervisor: Edit I Buzás; Katalin É Szabó-Taylor
E-mail addresses: buzas.edit@med.semmelweis-univ.hu; szabo-taylor.katalin@med.semmelweis-univ.hu

Bemutatás módja

Szóbeli

Kind

Szabad

Status

elfogadva

Accepted presentation method

szóbeli

Előadás fájl jóváhagyás

nem rendelkezett róla

Előadó

4685

Start

10:55

End

11:10

Authors (legacy)

Eszter Á Tóth; Semmelweis University, Department of Genetics, Cell- and Immunobiology; Budapest
Katalin É Szabó-Taylor; Semmelweis University, Department of Genetics, Cell- and Immunobiology; Budapest
Edit I Buzás; Semmelweis University, Department of Genetics, Cell- and Immunobiology; Budapest