PhD Scientific Days 2025

Poster Session III. - H: Pharmaceutical Sciences and Health Technologies

Proteomic, N-glycoproteomic and Chondroitin-sulfate Profiling of A549 and BEAS-2B Cell Line-derived Small Extracellular Vesicles

Előadó neve

Balbisi Mirjam, MSc

Neptun code

IG36TI

Előadó munkahelye

MTA-HUN-REN TTK Lendület (Momentum) Glycan Biomarker Research Group, HUN-REN Research Centre for Natural Sciences

Előadó telefonszáma

+36702822216

Előadó e-mail címe

balbisi.mirjam@phd.semmelweis.hu

Az előadás címe

Proteomic, N-glycoproteomic and Chondroitin-sulfate Profiling of A549 and BEAS-2B Cell Line-derived Small Extracellular Vesicles

Szerző(k) neve és munkahelye

Mirjam Balbisi¹, Tamás Langó², Virág Nikolett Horváth¹, Domonkos Pál¹, Gitta Schlosser³, Gábor Kecskeméti⁴, Zoltán Szabó⁴, Lilla Turiák¹

1: MTA-HUN-REN TTK Lendület (Momentum) Glycan Biomarker Research Group, HUN-REN Research Centre for Natural Sciences
2: Institute of Molecular Life Sciences, HUN-REN Research Centre for Natural Sciences
3: MTA-ELTE Lendület (Momentum) Ion Mobility Mass Spectrometry Research Group, ELTE Eötvös Loránd University
4: Department of Medical Chemistry, Albert Szent-Györgyi Medical School, University of Szeged

Bemutatás módja

Poszter

Szekció

Poster Session III. - H: Pharmaceutical Sciences and Health Technologies

Language of the presentation

English

Preferred session

Pharmaceutical Sciences and Health Technologies

Összefoglaló szövege

Introduction: Small extracellular vesicles (sEVs) are <200 nm particles secreted by cells that mediate intercellular communication and carry diverse biomolecules, including proteins. While protein content has been widely studied, post-translational modifications such as glycosylation remain less explored. Among glycosylated proteins, proteoglycans - composed of a core protein and glycosaminoglycan (GAG) chains like chondroitin sulfate (CS) - form a distinct class.
Aims: This study aimed to compare the proteomic, N-glycoproteomic, and CS GAG profiles of sEVs from A549 lung adenocarcinoma and BEAS-2B non-tumorigenic epithelial cell lines.
Methods: A549 and BEAS-2B cells were cultured for 3 days in F12 and BEGM media, respectively. sEVs were isolated by size-exclusion chromatography (n=6 per group), followed by solvent exchange and lysis. One part of each sample underwent tryptic digestion and N-glycopeptide enrichment via acetone precipitation. The other part was treated with chondroitinase ABC to release CS disaccharides. Peptides, N-glycopeptides, and CS disaccharides were analyzed by nanoUHPLC-MS(/MS).
Results: Of 945 proteins analyzed, 408 showed differential expression, including five CS proteoglycan core proteins. Versican and testican-1 were upregulated in A549, while aggrecan, syndecan-4 and CSPG4 were downregulated. Among the 301 N-glycoforms tested, 176 differed significantly between the sample groups. Dysregulated glycoforms included those of EV markers (e.g., galectin-3-binding protein) and proteoglycans (e.g., versican). CS disaccharide content was 3.4-fold higher in A549 sEVs, and the 6S/4S monosulfated disaccharide ratio decreased, suggesting altered sulfotransferase activity. Principal component analysis clearly separated tumor and non-tumor sEVs in all datasets, emphasizing that tumor and non-tumor sEVs not only differ at the protein level, but also have substantially different glycosylation patterns.
Conclusion: We demonstrated that A549 and BEAS-2B sEVs show distinct proteomic, N-glycoproteomic, and CS profiles, which calls for further research to understand the role of EV glycosylation in cancer.
Funding: Lendület (Momentum) Program of the Hungarian Academy of Sciences, Semmelweis 250+ Excellence PhD Scholarship

University

Semmelweis University

Supervisor

Lilla Turiák

Publication of my abstract

I give consent to the publication of my abstract on the website of the congress.

phd.section.field

in doctoral studies after complex exam (PhD)

Kind

Szabad

Status

elfogadva

Accepted presentation method

poszter

Előadás fájl jóváhagyás

nem rendelkezett róla

Előadó

7468

Start

14:42

End

14:48