N-Glycan profiling of lung adenocarcinoma in patients at different stages of disease

Abstract

Lung adenocarcinoma (LAC) is the most common form of lung cancer that increases in non-smokers at younger age. Altered protein glycosylation is one of the hallmarks of malignancy, its role in cancer progression is still poorly understood. In this study, we report mass spectrometric (MS) analysis of N-glycans released from fresh or defrosted tissue specimens from 24 patients with LAC. Comparison of cancerous versus adjacent healthy tissues revealed substantial differences in N-glycan profiles associated with disease. The significant increase in paucimannose and high-mannose glycans with 6-9 mannose residues and decline in the sialylated complex biantenary core fucosylated glycan with composition NeuAcGal 2 GlcNAc 2-Man 3 GlcNAc 2 Fuc were general features of tumors. In addition, 42 new N-glycan compositions were detected in cancerous tissues. The prominent changes in advanced disease stages were mostly observed in core fucosylated N-glycans with additional fucose (Fuc) residue/s and enhanced branching with non-galactosylated N-acetyl-glucosamine (GlcNAc) units. Both of these monosaccharide types were linked preferably on the 6-antenna. Importantly, as compared with noncancerous tissues, a number of these significant changes were clearly detectable early on in stage I. Application of N-glycan data obtained from tissues was next assessed and validated for evaluation of small sized biopsies obtained via bronchoscopy. In summary, observed alterations and data of newly detected N-glycans expand knowledge about the glycosylation in LAC and may contribute to research in more tailored therapies. Moreover, the results demonstrate effectiveness of the presented approach for utility in rapid discrimination of cancerous from healthy lung tissues.

Comprehensive N-glycosylation mapping of envelope glycoprotein from tick-borne encephalitis virus grown in human and tick cells

“…From the first appearance of eluate from the cartridge, the eluates were collected and analyzed. When using the digest from whole cell lysates, the XC-eluates were immediately loaded onto graphitized carbon cartridges (1 ml tubes, Supelco) and the same protocol was followed as described previously 29 . Glycan fractions were combined, evaporated and analyzed by MALDI-MS.…”

In vivo and in vitro cell based model of lung adenocarcinoma from patients with pleural effusion

“…Frequently, observations obtained from in vitro studies are subsequently validated in vivo. We have recently reported an effective procedure for N-glycan profiling of tissue specimens which was first developed using in vitro established tumor cell lines and then was successfully applied for fresh tumor specimens obtained from patients with LAC [50,52].…”

Mass Spectrometry Imaging of N-Glycans Reveals Racial Discrepancies in Low Grade Prostate Tumors

“…Specifically, prostate tumors exhibit accumulation of highmannose glycans which increase with tumor grade group (Figure 3A-C), a common feature of human cancers that correlates with more aggressive cancer phenotypes (20,21). Accumulation of high mannose N-glycans in prostate tumors suggests a lack of N-glycan trimming reactions and a decrease in mannosidase activity, or aberrant mannose metabolism (50)(51)(52). Additionally, we found prostate tumors accumulate tri-and tetra-antennary complex glycans containing a core fucose moiety ( Figure 3D-F), suggesting prostate tumors have enhanced GlcNAc metabolism.…”