Diagnostic Potential of Plasma IgA1 O-Glycans in Discriminating IgA Nephropathy From Other Glomerular Diseases and Healthy Participants

Background: Aberrant O-glycosylation of IgA1 plays an important role in IgA nephropathy pathogenesis. Previous proteomic studies analyzed O-glycans of the circulating IgA1 hinge region and found that the N-acetylgalactosamine (GalNAc) and galactose numbers in the hinge region of IgA1 of patients with IgA nephropathy were lower than those in healthy participants. However, the diagnostic performance of the O-glycosylation traits in the hinge region of plasma IgA1 for IgA nephropathy remains unelucidated. The present study aimed to determine the difference in plasma IgA1 hinge region O-glycoforms among IgA nephropathy, non-IgA nephropathy disease controls, and healthy participants, and to further evaluate the diagnostic performance of plasma IgA1 glycosylation traits. Methods: Sixty-two patients with biopsy-proven primary IgA nephropathy, 30 age- and sex-matched non-IgA nephropathy disease controls (10 patients with membranous nephropathy, 10 with focal segmental glomerulosclerosis, and 10 with minimal change disease), and 30 healthy participants were prospectively recruited. Plasma galactose deficient-IgA1 levels were measured using a KM55 kit. Plasma IgA was extracted using IgA immunoaffinity beads. After de-N-glycosylation, reduction, alkylation, trypsin digestion, and O-glycopeptide enrichment via hydrophilic interaction liquid chromatography, liquid chromatography tandem mass spectrometry (LC-MS/MS) was applied to analyze the IgA1 O-glycosylation patterns and we derived the plasma IgA1 O-glycosylation traits. Results: Plasma IgA1 O-glycosylation patterns were significantly changed in IgA nephropathy patients compared to those with non-IgA nephropathy disease controls and healthy participants. The GalNAc number was lowest in IgA nephropathy patients. In addition, a similar result was observed for the galactose number in the IgA1 hinge region. These values showed moderate potential for discriminating between IgA nephropathy and the controls. When these values were combined, the area under the curve increased compared to when they were considered individually. When further adding a clinical indicator, the area under the curve of the GalNAc-galactose-IgA panel exceed 0.9 in discriminating IgA nephropathy from the controls. Conclusion: The amount of GalNAc and galactose in plasma IgA1 hinge region identified by glycoproteomics could be used as a diagnostic biomarker for IgA nephropathy. The panel containing GalNAc, galactose, and circulating IgA displayed excellent diagnostic performance and is promising for practical clinical applications.

Automated summary

This study demonstrated the potential diagnostic value of O-glycoforms in the hinge region of plasma IgA1 for IgA nephropathy. The levels of GalNAc and galactose were significantly different in IgA nephropathy patients compared to controls, and combining these values improved the diagnostic performance. A panel containing GalNAc, galactose, and circulating IgA showed excellent diagnostic potential.