PhD Scientific Days 2023

Molecular Sciences III.

Deciphering the molecular landscape of the intraluminal or plasma membrane origin of extracellular vesicles and exomere.

Előadó neve

Ghosal, Sayam

Neptun code

ZK30IU

Előadó munkahelye

HCEMM EV Research Group, Department of Genetics, Cell and Immunobiology, Semmelweis University

Előadó telefonszáma

06300164496

Előadó e-mail címe

ghosal.sayam@gmail.com

Az előadás címe

Deciphering the molecular landscape of the intraluminal or plasma membrane origin of extracellular vesicles and exomere.

Szerző(k) neve és munkahelye

Sayam Ghosal, HCEMM-SU Extracellular Vesicles Research Group, Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest.

Brachyahu Meir Kestecher, HCEMM-SU Extracellular Vesicles Research Group, Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest.

Bernadett Réka Bodnár, HCEMM-SU Extracellular Vesicles Research Group, Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest.

Adrienn Szabó, HCEMM-SU Extracellular Vesicles Research Group, Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest.

Edit Buzás, HCEMM-SU Extracellular Vesicles Research Group, Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest.

Xabier Osteikoetxea, HCEMM-SU Extracellular Vesicles Research Group, Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest.

Bemutatás módja

Szóbeli

Szekció

Molecular Sciences III.

Language of the presentation

English

Preferred session

Molecular Sciences

Összefoglaló szövege

Introduction:
Extracellular vesicles (EVs) are a heterogeneous group of membrane-bound structures released by various cell types that carry diverse cargo, such as proteins, lipids, and nucleic acids. Over the past few decades, EVs have attracted significant attention in the field of cell biology due to their essential role in intercellular communication and their potential diagnostic and therapeutic applications. Recent studies have revealed the heterogeneity of EVs, which vary in size, biogenesis, and cargo. Moreover, a novel subpopulation of extracellular nanoparticles, called exomeres, has been identified, but their biogenesis and release mechanisms remain unclear. Understanding the precise molecular mechanisms underlying the formation and release of different EV subpopulations is crucial to harness the full potential of EVs for biomedical applications.
Aim:
In this study, we are aiming to find definitive molecular markers of the different biogenetic routes of EVs formation, such as large EVs (diameter ≥ 1 µm), medium-sized EVs (200-800 nm), small EVs (50-150 nm) and recently discovered extracellular nanoparticles, exomere (30-70 nm).
Method:
We used molecular cloning to design secreting cells to express recognized molecular markers of the plasma membrane, EV, lysosome, and autophagosomes to get a better understanding of the cellular origin of diverse subpopulations of EVs and exomere. We have isolated EVs and exomere from Hek293T cells using the differential ultracentrifugation and filtration method following MISEV2018 guidelines. We characterized these EVs with nanoparticle tracking analysis (NTA), protein assay, lipid assay, super-resolution flow cytometry, and immune gold transmission electron microscopy (TEM). Here, direct stochastic optical reconstruction microscopy (dSTORM) was employed to visualize EVs in three dimensions and to localize molecule clusters.
Results:
The mean size of the mEVs and sEVs is 210 nm and 120 nm, and the size distribution ranges from 100-500 nm and 110-160 nm, respectively. The concentration of sEVs expressing Syntenin1, Arf6 and TSPAN14 was significantly higher compared to sEVs expressing LC3b. It was evident that the CD63 gave higher eGFP positive events in both mEVs and exomere. Interestingly, the protein-to-lipid ratio was considerably higher among the exomeres compared to EV sub-populations.
Conclusion:
Together, these traits indicate potentially significant variations in the cellular origin of different EV subpopulations, particularly for exomere. Overall, research into the biogenesis of exomere is still ongoing, and further investigations are required to completely comprehend the underlying processes.
Funding
European Union’s Horizon 2020 Research and Innovation Programme under grant agreement No 739593, 2019-2.1.7-ERA-NET-2021-00015, STIA-KFI-2022, RRF-2.3.1-21-2022-00003, TKP2021-EGA-23, ELKH-SE Extracellular Vesicle Research Group grant.

University and Doctoral School

Semmelweis University, Doctoral School of Molecular Medicine

Supervisor

Xabier Osteikoetxea

Publication of my abstract

I do not give consent to the publication of my abstract on the website of the congress.

Kind

Szabad

Status

elfogadva

Accepted presentation method

szóbeli

Előadás fájl jóváhagyás

nem rendelkezett róla

Előadó

7476

Start

12:30

End

12:45

Authors (legacy)

Sayam Ghosal, HCEMM-SU Extracellular Vesicles Research Group, Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest.

Brachyahu Meir Kestecher, HCEMM-SU Extracellular Vesicles Research Group, Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest.

Bernadett Réka Bodnár, HCEMM-SU Extracellular Vesicles Research Group, Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest.

Adrienn Szabó, HCEMM-SU Extracellular Vesicles Research Group, Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest.

Edit Buzás, HCEMM-SU Extracellular Vesicles Research Group, Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest.

Xabier Osteikoetxea, HCEMM-SU Extracellular Vesicles Research Group, Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest.