Interlaboratory evaluation of plasma N-glycan antennary fucosylation as a clinical biomarker for HNF1A-MODY using liquid chromatography methods

Abstract

Antennary fucosylation alterations in plasma glycoproteins have been previously proposed and tested as a biomarker for differentiation of maturity onset diabetes of the young (MODY) patients carrying a functional mutation in the HNF1A gene. Here, we developed a novel LC-based workflow to analyze blood plasma N-glycan fucosylation in 320 diabetes cases with clinical features matching those at risk of HNF1A-MODY. Fucosylation levels measured in two independent research centers by using similar LC-based methods were correlated to evaluate the interlaboratory performance of the biomarker. The interlaboratory study showed good correlation between fucosylation levels measured for the 320 cases in the two centers with the correlation coefficient (r) of up to 0.88 for a single trait A3FG3S2. The improved chromatographic separation allowed the identification of six single glycan traits and a derived antennary fucosylation trait that were able to differentiate individuals carrying pathogenic mutations from benign or no HNF1A mutation cases, as determined by the area under the curve (AUC) of up to 0.94. The excellent (r = 0.88) interlaboratory performance of the glycan biomarker for HNF1A-MODY further supports the development of a clinically relevant diagnostic test measuring antennary fucosylation levels.

Development of an exoglycosidase plate-based assay for detecting α1-3,4 fucosylation biomarker in individuals with HNF1A-MODY

“… Correlations between N -glycan antennary fucosylation levels measured as indexes by LC method in the previous study ( Demus et al. 2021 ) and α1-3,4 fucosylation levels measured by the exoglycosidase plate-based assay for 31 HNF1A-MODY positive cases (A), and 320 diabetes cases including 31 HNF1A-MODY positive and 289 negative cases (B).…”

“…The exoglycosidase plate-based assay developed in this work is based on the use of a novel α1-3,4 specific fucosidase (E1_10125), which is characterized by the ability to remove fucose residues from glycoconjugates present in blood plasma ( Wu et al. 2020 ; Demus et al. 2021 ).…”

“…Previously, we reported significant correlations between C-reactive protein (CRP) levels and antennary fucosylation levels, which were measured using antennary fucosylated N -glycan traits by an LC-based method ( Demus et al. 2021 ).…”

“…We next compared the fucosylation indexes previously measured using an LC-based method ( Demus et al. 2021 ), which reflect the levels of antennary fucosylation in the full plasma N -glycome, with the α1-3,4 fucosylation levels in blood plasma samples measured by the exoglycosidase plate-based assay.…”

“…2013 ; Juszczak et al. 2019 ) and we recently showed the excellent interlaboratory performance of the N -glycan biomarker using liquid chromatography (LC) methods ( Demus et al. 2021 ).…”