The Use of the Neoglycolipid-Based Oligosaccharide Microarray System in the Diagnosis of Endometriosis – Preliminary Study

Abstract

Objective

Endometriosis presents diagnostic challenges, and there is a need for developing novel biomarkers with satisfactory specificity and sensitivity. Glycomics, exploring glycosylation changes in glycoproteins, offers potential solutions. The aim of this study was to analyze the carbohydrate-binding properties of IgG and IgM antibodies in the plasma and peritoneal fluid samples and to identify any differences in the presence and the specificities of anti-carbohydrate antibodies in the endometriosis patient and the controls.

Methods

Multicenter study was conducted in Poland between 2018 and 2019. Plasma and peritoneal fluid samples were collected from women undergoing laparoscopic surgery. Endometriosis patients (n=8) and controls (n=8), matched for cycle phase and disease stage, were selected. The neoglycolipid-based oligosaccharide microarray system was used to investigate IgG and IgM antibody binding properties to glycan-related probes in biological materials.

Results

In peritoneal fluid samples, IgM binding to the following probes was significantly higher in endometriosis: GSC-915-4 (new), LNFP-I, NeuAcα-(6’)LNnO (F1), B-like decaosylceramide, log10(GM1-penta), and log10(GSC-915-5). In a control group higher IgG binding to log10(Orsay-5-AO) was observed. In plasma samples, endometriosis showed higher IgG binding to log10(NeuAcα-(6’)LNnO (F1)) and lower IgG binding to Gal2GlcNAc(1-3)-AO. After Benjamin–Hochberg correction, differences were not significant. Effect sizes highlighted some glycan probes in both plasma and peritoneal fluid. Strong correlations were observed among binding to certain glycan probes.

Conclusion

This preliminary study suggests glycomics’ potential contribution to endometriosis diagnosis and understanding of its pathophysiology. Neoglycolipid-based microarrays hold promise for non-invasive endometriosis diagnostic tools. Further investigations with larger cohorts are warranted to validate these findings and explore potential correlations with antibody levels in plasma and peritoneal fluid. Glycomics emerges as a valuable diagnostic asset in endometriosis research.

Keywords:

• endometriosis
• glycomics
• diagnosis
• microarray

Abbreviations

AO, aminooxy; BMI, body mass index; DHPE, 1,2-dihexadecyl-sn-glycero-3-phosphoethanolamine; EDTA, ethylenediaminetetraacetic acid; P, plasma; PF, peritoneal fluid; NGLs, neoglycolipids.

Institutional Review Board Statement

The Bioethics Committee operating at the Medical University of Warsaw approved the collection of the material in accordance with opinion No. KB 223/2017, issued on December 12, 2017. The Bioethics Committee operating at the Calisia University approved the method of analysis of the previously collected material using neoglycolipid-based oligosaccharide microarray system (opinion 04/2021, issued on December 20, 2021).

Informed Consent Statement

Informed consent was obtained from all subjects involved in the study.

Data Sharing Statement

The data presented in this study are available on request from the corresponding author.

Acknowledgments

We would like to thank our collaborators from the MZ project for their participation in the recruitment of selected samples for this project. We would also like to thank the Glycosciences Laboratory team for conducting glycomics analysis.

Author Contributions

Cezary Wojtyła was responsible for study conception, study design, acquisition, analysis, interpretation, and visualization of the data. He acquired financial support for the project leading to this publication, and he was responsible for managing and coordinating the planning and implementation of research activities, selecting patients and material for this project, and analysis. He wrote and edited the manuscript. Ignacy Tołwiński was responsible for study conception, analysis, and interpretation of the data, and he was also responsible for data visualization and writing the manuscript. Piotr Laudański was responsible for study conception and study design. He acquired financial support for the project leading to this publication. He was responsible for organizing the collection of biological material, editing the manuscript, and critical revision. All authors have agreed on the journal to which the article will be submitted. All authors have reviewed and agreed on all versions of the article. All authors have agreed to take responsibility and be accountable for the contents of the article.

Disclosure

The authors report no conflicts of interest in this work.

Additional information

Funding

This study was supported by grants from National Science Centre (Narodowe Centrum Nauki - MINIATURA 5, grant no. 2021/05/X/NZ1/00646), Polish Ministry of Health (grant no. 6/6/ 4/1/NPZ/2017/1210/1352) and MSCA-RISE-2020 project TRENDO (grant no. 101008193).