Dataset Information

Role of N-glycosylation in FcRIIIa interaction with IgG

ABSTRACT: Immunoglobulins G (IgG) and their Fc gamma receptors (FcγRs) play important roles in our immune system. The conserved N-glycan in the Fc region of IgG1 impacts interaction of IgG with FcγRs and the resulting effector functions, which has led to design of antibody therapeutics with greatly improved antibody-dependent cell cytotoxicity (ADCC) activities. Studies have suggested that also N-glycosylation of the FcγRIII affects receptor inteactions with IgG, but detailed studies of the interaction of IgG1 and FcγRIIIa with distinct N-glycans have been hindered by the natural heterogeneity in N-glycosylation. In this study, we employed comprehensive genetic engineering of the N-glycosylation capacities in mammalian cell lines to express IgG1 and FcγRIIIa with different N-glycan structures to more generally explore the role of N-glycosylation in IgG1:FcRIIIa binding interactions. We included FcRIIIa variants of both the 158F and 158V allotypes and investigated the key N-glycan features that affected binding affinity. Our study confirms that afucosylated IgG1 has the highest binding affinity to oligomannose FcγRIIIa, a glycan structure commonly found on FcγRIIIa expressed by NK cells but not monocytes or recombinantly expressed FcγRIIIa.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human) Mus Musculus (mouse)

TISSUE(S): Cell Culture

SUBMITTER: Sergey Vakhrushev

LAB HEAD: Sergey Vakhrushev

PROVIDER: PXD035846 | Pride | 2022-08-30

REPOSITORIES: Pride

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			Action	DRS
	200820_Lumos__LC1200__nanoFlex_Julie_Hek293_FcgR3A_F_site_specific_90min_HCD_48_Chy_GluC_2ul.raw	Raw
	200820_Lumos__LC1200__nanoFlex_Julie_Hek293_FcgR3A_F_site_specific_90min_HCD_48_PNGaseF_Chy_GluC_2ul.raw	Raw
	200820_Lumos__LC1200__nanoFlex_Julie_Hek293_FcgR3A_F_site_specific_90min_HCD_49_Chy_GluC_2ul.raw	Raw
	200820_Lumos__LC1200__nanoFlex_Julie_Hek293_FcgR3A_F_site_specific_90min_HCD_49_PNGaseF_Chy_GluC_2ul.raw	Raw
	200820_Lumos__LC1200__nanoFlex_Julie_Hek293_FcgR3A_F_site_specific_90min_HCD_50_Chy_GluC_2ul.raw	Raw

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Project description:BACKGROUND: Novel biomarkers are urgently needed to distinguish between benign and malignant thyroid nodules and detect thyroid cancer in the early stage. The associations between serum IgG N-glycosylation and thyroid cancer risk have been revealed. We aimed to explore the potential of IgG N-glycan traits as biomarkers in the differential diagnosis of thyroid cancer.METHODS: Plasma IgG N-glycome analysis was applied to a discovery cohort followed by independent validation. IgG N-glycan profiles were obtained using a robust quantitative strategy based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. IgG N-glycans were relatively quantified, and classification performance was evaluated based on directly detected and derived glycan traits.RESULTS: Four directly detected glycans were significantly changed in thyroid cancer patients compared to that in non-cancer controls. Derived glycan traits and a classification glycol-panel were generated based on the directly detected glycan traits. In the discovery cohort, derived trait BN (bisecting type neutral N-glycans) and the glyco-panel showed potential in distinguishing between thyroid cancer and non-cancer controls with AUCs of 0.920 and 0.917, respectively. The diagnostic potential was further validated. Derived trait BN and the glycol-panel displayed “accurate” performance (AUC>0.8) in discriminating thyroid cancer from benign thyroid nodules and healthy controls in the validation cohort. Meanwhile, derived trait BN and the glycol-panel also showed diagnostic potential in detecting early-stage thyroid cancer.CONCLUSIONS: IgG N-glycome analysis revealed N-glycomic differences that allow classification of thyroid cancer from non-cancer controls. Our results suggested that derived trait BN and the classification glyco-panel rather than single N-glycans may serve as candidate biomarkers for further validation.

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Role of N-glycosylation in FcRIIIa interaction with IgG

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