Related Experiment Video
Updated: Sep 6, 2025
Glycomics-Guided Glycoproteomics Facilitates Comprehensive Profiling of the Glycoproteome in Complex Tumor Microenvironments
Published on: February 7, 2025
An Integrated Mass Spectrometry-Based Glycomics-Driven Glycoproteomics Analytical Platform to Functionally
Michael Russelle S Alvarez1,2, Qingwen Zhou2, Sheryl Joyce B Grijaldo1
1Institute of Chemistry, College of Arts and Sciences, University of the Philippines Los Baños, Los Baños 4031, Philippines.
Abstract:
Cancer progression is linked to aberrant protein glycosylation due to the overexpression of several glycosylation enzymes. These enzymes are underexploited as potential anticancer drug targets and the development of rapid-screening methods and identification of glycosylation inhibitors are highly sought. An integrated bioinformatics and mass spectrometry-based glycomics-driven glycoproteomics analysis pipeline was performed to identify an N-glycan inhibitor against lung cancer cells. Combined network pharmacology and in silico screening approaches were used to identify a potential inhibitor, pictilisib, against several glycosylation-related proteins, such as Alpha1-6FucT, GlcNAcT-V, and Alpha2,6-ST-I. A glycomics assay of lung cancer cells treated with pictilisib showed a significant reduction in the fucosylation and sialylation of N-glycans, with an increase in high mannose-type glycans. Proteomics analysis and in vitro assays also showed significant upregulation of the proteins involved in apoptosis and cell adhesion, and the downregulation of proteins involved in cell cycle regulation, mRNA processing, and protein translation. Site-specific glycoproteomics analysis further showed that glycoproteins with reduced fucosylation and sialylation were involved in apoptosis, cell adhesion, DNA damage repair, and chemical response processes. To determine how the alterations in N-glycosylation impact glycoprotein dynamics, modeling of changes in glycan interactions of the ITGA5-ITGB1 (Integrin alpha 5-Integrin beta-1) complex revealed specific glycosites at the interface of these proteins that, when highly fucosylated and sialylated, such as in untreated A549 cells, form greater hydrogen bonding interactions compared to the high mannose-types in pictilisib-treated A549 cells. This study highlights the use of mass spectrometry to identify a potential glycosylation inhibitor and assessment of its impact on cell surface glycoprotein abundance and protein-protein interaction.
Insights
Researchers identified pictilisib as a potential lung cancer drug by targeting aberrant protein glycosylation. This inhibitor alters N-glycans, impacting cell adhesion and apoptosis pathways, offering a new strategy against cancer progression.
Area of Science:
- Biochemistry
- Glycobiology
- Cancer Research
Background:
- Aberrant protein glycosylation is a hallmark of cancer progression, driven by overexpressed glycosylation enzymes.
- Glycosylation enzymes represent underexploited anticancer drug targets, necessitating rapid screening and inhibitor identification.
Purpose of the Study:
- To identify an N-glycan inhibitor for lung cancer cells using an integrated bioinformatics and mass spectrometry-based glycomics-driven glycoproteomics approach.
- To investigate the impact of a potential inhibitor on cellular glycosylation, protein expression, and protein-protein interactions.
Main Methods:
- Integrated bioinformatics, network pharmacology, and in silico screening to identify potential glycosylation inhibitors.
- Mass spectrometry-based glycomics and glycoproteomics to analyze N-glycan changes and site-specific glycosylation alterations.
- In vitro assays and protein-protein interaction modeling (ITGA5-ITGB1 complex) to assess functional impacts.
Main Results:
- Pictilisib identified as a potential inhibitor targeting Alpha1-6FucT, GlcNAcT-V, and Alpha2,6-ST-I.
- Pictilisib treatment significantly reduced N-glycan fucosylation and sialylation, increasing high mannose-type glycans in lung cancer cells.
- Altered glycosylation impacted apoptosis, cell adhesion, DNA repair, and cell cycle regulation pathways, with specific effects on the ITGA5-ITGB1 complex interactions.
Conclusions:
- Mass spectrometry-based glycomics and glycoproteomics can effectively identify glycosylation inhibitors and assess their impact on cancer cells.
- Pictilisib demonstrates potential as an anticancer agent by modulating N-glycosylation and downstream cellular processes.
- Understanding glycosylation dynamics is crucial for developing targeted cancer therapies and predicting drug efficacy.
