Mass spectrometry-based characterization of proteomic and glycosylation alterations in lung cancer tissues and extracellular vesicles
Balbisi Mirjam
Gyógyszertudományok és Egészségügyi Technológiák Tagozat
Dr. Zelkó Romána
SE Semmelweis Szalon
2026-09-02 11:00:00
Modern Trends in Pharmaceutical Scientific Research
Dr. Antal István
Dr. Turiák Lilla
Dr. Ozohanics Olivér
Dr. Márk László
Dr. Tábi Tamás
Dr. Dunkel Petra
Dr. Imre Tímea
Lung cancer is characterized by complex molecular alterations that go beyond changes in protein expression and also include alterations in glycosylation. Among these,
N-glycosylation and GAGs play essential roles in cell signaling, extracellular matrix organization, and tumor progression. This thesis aimed to investigate these molecular features using advanced HPLC-MS-based analytical approaches, combining proteomics with in-depth glycosylation analysis in two complementary biological systems.
The first part of the work focused on spatially resolved lung cancer tissues carrying ALK rearrangement. By applying on-surface digestion techniques, both protein composition as well as CS/DS and HS GAG disaccharide profiles were analyzed from morphologically distinct regions. The results revealed clear molecular differences between tumor and adjacent non-tumor regions, particularly in pathways related to extracellular matrix organization and protein biosynthesis. In parallel, GAG analysis showed an increased overall abundance and altered sulfation patterns in tumor regions. Between tumor subtypes, the most prominent dysregulated processes were associated with protein synthesis, and the largest differences were observed in the total amount of HS disaccharides.
The second study investigated sEVs derived from A549 lung adenocarcinoma and
BEAS-2B non-tumorigenic epithelial cell lines. Comprehensive characterization included proteomic, glycomics guided N-glycoproteomic, and CS/DS GAG analyses. The results demonstrated that all three molecular profiles clearly distinguished the two sEVs types. Differential expression of several PG core proteins was observed, and they showed mixed patterns. Several proteins, including laminins, galectin-3-binding protein, and versican, exhibited multiple N-glycosylation alterations. Additionally, GAG analysis revealed an increased total amount of CS disaccharides along with a shift toward higher 4S sulfation in A549 sEVs.
Overall, this work highlights the importance of combining proteomic and glycosylation approaches to achieve a more comprehensive understanding of lung cancer biology. These results contribute to the growing field of cancer glycobiology and support the potential application of glycosylation patterns in future biomarker discovery and disease characterization.
