Extended coverage of human serum glycosphingolipidome by 4D-RP-LC TIMS-PASEF unravels association with Parkinson’s disease

• Published in: Nature communications Vol. 16; no. 1; pp. 4567 - 22
• Main Authors: Vo, Huong Giang, Gonzalez-Escamilla, Gabriel, Mirzac, Daniela, Rotaru, Lilia, Herz, Damian, Groppa, Sergiu, Bindila, Laura
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 16.05.2025; Nature Publishing Group; Nature Portfolio
• Subjects: 101/58; 631/1647/296; 631/45/608; 692/53; 692/617/375/1718; Adult; Aged; Annotations; Chromatography; Chromatography, Liquid - methods; Chromatography, Reverse-Phase - methods; Disease; Female; Fractionation; Glycosphingolipids; Glycosphingolipids - blood; Glycosphingolipids - chemistry; Heterogeneity; Humanities and Social Sciences; Humans; Lipidomics - methods; Lysosomal storage diseases; Male; Mass Spectrometry - methods; Middle Aged; Monosaccharides; Movement disorders; multidisciplinary; Neurodegenerative diseases; Parkinson Disease - blood; Parkinson Disease - diagnosis; Parkinson's disease; Phenotyping; Science; Science (multidisciplinary); Scientific imaging; Subgroups
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-025-59755-6

Glycosphingolipids (GSLs) are important targets in immune, infectious, lysosomal storage diseases, cancer, and neurodegenerative diseases. Circulatory GSLs profiling in clinical samples is restricted by the lack of mid- and high-throughput analytical methods and deep coverage of long-chain sialylated glycosphingolipidome. We present a 4-dimensional (4D)-glycosphingolipidomics platform for routine glycosphingolipidome profiling encompassing: extraction and fractionation of sialylated GSLs with 3 to 15 monosaccharides, neutral GSLs and sulfatides; µL-flow reversed-phase LC-TIMS-PASEF MS analysis; semi-quantification strategy adapted for fractionated glycosphingolipidome, and referential CCS, RT, and m / z values for GSLs annotation. 4D-glycosphingolipidomics of human serum reveals a high structural heterogeneity, amounting to 376 GSLs: 159 GSLs of ganglio- and neolacto-series, 145 neutral GSLs and 72 sulfatides. Here we demonstrate the platform’s utility for clinical profiling of Parkinson’s disease (PD) sera. 41 neolacto- and ganglio-species discriminate PD patients from controls and 14 GSLs differentiate sex subgroups, laying the foundation for further functional GSL studies with PD. 4D-TIMS-glycosphingolipidomics expedite clinical serum profiling of 376 GSLs covering ganglio- and neolacto-series, neutrals and sulfatides. Clinical phenotyping of Parkinson’s disease cohort (sub)groups identified up to 41 GSLs fingerprints.