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To establish specific structural evidence that glycans are indeed blunted/altered from the glycan profiling of glycan moieties on the surface of leukocytes

ABSTRACT: We want to characterize the glycan moieties on the surface of leukocytes treated with fluorinated analogs of glucosamine. Since these metabolic inhibitors compete with natural-occuring glucosamine used in oligosaccharide chain formation, these compounds have been shown to be effective in lowering Type 2 lactosamine and sialylated Lewis antigens, which results in altered selectin ligand functions.

ORGANISM(S): Homo sapiens

PROVIDER: GSE32556 | GEO | 2011/10/03

SECONDARY ACCESSION(S): PRJNA147025

REPOSITORIES: GEO

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To establish specific structural evidence that glycans are indeed blunted/altered from the glycan profiling of glycan moieties on the surface of leukocytes

Project description:We want to characterize the glycan moieties on the surface of leukocytes treated with fluorinated analogs of glucosamine. Since these metabolic inhibitors compete with natural-occuring glucosamine used in oligosaccharide chain formation, these compounds have been shown to be effective in lowering Type 2 lactosamine and sialylated Lewis antigens, which results in altered selectin ligand functions. We need to establish specific structural evidence that glycans are indeed blunted/altered. Current research efforts include investigations on how circulating leukocytes traffic to inflamed tissues, how circulating solid tumor cells interact with endothelial linings to enhance metastatic potential, and how treatment with analogs of glucosamine disrupt leukocyte trafficking mechanisms and/or influence effector/regulatory function of T cells.

2011-10-02 | E-GEOD-32556 | biostudies-arrayexpress

E-selectin Affinity Glycoproteomics Reveals Neuroendocrine Proteins and the Secretin Receptor as a Poor Prognosis Signature in Colorectal Cancer

Project description:Colorectal cancer (CRC) cells express glycoproteins with sialylated Lewis antigens (sLe), crucial for metastasis via E-selectin binding. However, these glycoepitopes lack cancer specificity, and E-selectin-targeted glycoproteins are unknown. Here we established a framework for identifying metastasis-linked glycoproteoforms for precision oncology. We found that over 70% of colorectal tumours overexpressed sLeA/X, yet without association with metastasis or survival. This prompted deeper exploration on the sialoglycoproteome of metastatic tumours. Nearly 600 glycoproteins were identified using E-selectin affinity enrichment and mass spectrometry, significantly broadening our understanding of potential metastasis-related glycoproteins. This glycoproteome was linked to cell adhesion, active transport of molecules and ions across the plasma membrane, oncogenic pathways, angiogenesis, and, unexpectedly, neuroendocrine functions. Employing an in-house algorithm for targetability prioritization, the less studied secretin receptor (SCTR) emerged as a top-ranked glycoprotein. The screening of tumours confirmed SCTR's association with poor prognosis and metastasis. N-glycosylation carrying sLe antigens added cancer specificity to SCTR. Prognostic links were reinforced by TCGA-based investigations. In summary, SCTR, a relatively unknown CRC glycoprotein, holds potential as a biomarker of poor prognosis and as a E-selectin ligand, suggesting an unforeseen role in metastasis warranting confirmation. Future investigations should also focus on this glycoproteoforms’ biological foreseeing clinical applications.

2024-11-11 | PXD054727 | Pride

Immune cells surveil aberrantly sialylated O-glycans on megakaryocytes to regulate platelet count

Project description:Immune thrombocytopenia (ITP) is a common platelet disorder in pediatric patients. Pediatric and adult ITP have been associated with sialic acid alterations, but the pathophysiology of ITP remains elusive, and ITP is often a diagnosis of exclusion. Our analysis of pediatric ITP plasma samples showed increased anti-Thomsen-Friedenreich antigen (TF-antigen) antibody representation, suggesting increased exposure of the typically sialylated and cryptic TF-antigen in these patients. The O-glycan sialyltransferase St3gal1 add sialic acid specifically on the TF-antigen. To understand if TF-antigen exposure associates with thrombocytopenia, we generated a mouse model with targeted deletion of St3gal1 in megakaryocytes (MK) (St3gal1MK-/-). TF-antigen exposure was restricted to MKs and resulted in thrombocytopenia. Deletion of Jak3 in St3gal1MK-/- mice normalized platelet counts implicating involvement of immune cells. Interferon-producing Siglec H-positive bone marrow (BM) immune cells engaged with O-glycan sialic acid moieties to regulate type I interferon (IFN-I) secretion and platelet release (thrombopoiesis), as evidenced by partially normalized platelet count following and inhibition of interferon and Siglec H receptors. Single cell RNAseq determined that TF-antigen exposure by MKs primed St3gal1MK-/- BM immune cells to release IFN-I. Single cell RNAseq further revealed a new population of immune cells with a plasmacytoid dendritic cell (pDC)-like signature and concomitant upregulation of immunoglobulin re-arrangement gene transcripts Igkc and Ighm, suggesting additional immune regulatory mechanisms. Thus, aberrant TF-antigen moieties, often found in pathological conditions, regulate immune cells and thrombopoiesis in the BM, leading to reduced platelet count.

2021-06-29 | GSE179067 | GEO

To determine the changes in the expression of human colon cancer cells (HCT-15) untreated and treated with 5-aza-2''-deoxycytidine

Project description:Human colon cancer cells (HCT-15) treated with 5-aza-2'-deoxycytidine greatly enhanced sialyl Lewis X epitope, which correlated with increased binding of these cells to immobilized E-selectin in a parallel plate dynamic flow binding assay. Human colon cancer cells (HCT-15) treated with 5-aza-2'-deoxycytidine greatly enhanced sialyl Lewis X epitope, which correlated with increased binding of these cells to immobilized E-selectin in a parallel plate dynamic flow binding assay. The sialyl Lewis x epitope resides at a 250 Kd protein band as assessed following immunoprecipitation. We would like to know changes in the expression of glycogenes following 5-aza-2'-deoxycytidine treatment.

2011-09-05 | E-GEOD-31933 | biostudies-arrayexpress

Short-term effects of human versus bovine sialylated milk oligosaccharide microinjection on zebrafish larvae survival, locomotor behavior and gene expression

Project description:Milk oligosaccharides are a complex class of carbohydrates that act as bioactive factors in numerous defensive and physiological functions, including brain development. Early nutrition can modulate nervous system development and can lead to epigenetic imprinting. We attempted to increase the sialylated oligosaccharide content of zebrafish yolk reserves, with the aim of evaluating any short-term effects of the treatment on mortality, locomotor behavior, and gene expression. Wild-type embryos were microinjected with saline solution or solutions containing sialylated milk oligosaccharides extracted from human and bovine milk. The results suggest that burst activity and larval survival rates were unaffected by the treatments. Locomotion parameters were found to be similar during the light phase between control and treated larvae; in the dark, however, milk oligosaccharide-treated larvae showed increased test plate exploration. Thigmotaxis results did not reveal significant differences in either the light or the dark conditions. RNA-seq analysis indicated that both treatments exert an antioxidant effect in developing fish. Moreover, sialylated human milk oligosaccharides seemed to increase the expression of genes related to cell cycle control and chromosomal replication, while bovine-derived oligosaccharides caused an increase in the expression of genes involved in synaptogenesis and neuronal signaling. These data shed some light on this poorly explored research field, showing that both human and bovine oligosaccharides support brain proliferation and maturation.

2023-04-04 | GSE218857 | GEO

To determine the changes in the expression of human colon cancer cells (HCT-15) untreated and treated with 5-aza-2''-deoxycytidine

2011-09-06 | GSE31933 | GEO

Sialylation of N-linked glycans influences the immunomodulatory effects of IgM on T cells

Project description:Human serum IgM antibodies are composed of heavily glycosylated polymers with five glycosylation sites on the μ (heavy) chain and one glycosylation site on the J chain. In contrast to IgG glycans, which are vital for a number of biological functions, virtually nothing is known about structure-function relationships of IgM glycans. Natural IgM is the earliest immunoglobulin produced and recognizes multiple antigens with low affinity, whilst immune IgM is induced by antigen exposure and is characterized by a higher antigen specificity. Natural anti-lymphocyte IgM is present in the serum of healthy individuals and increases in inflammatory conditions. It is able to inhibit T cell activation, but the underlying molecular mechanism is not understood. Here we show, for the first time, that sialylated N-linked glycans induce the internalization of IgM by T cells, which in turn causes severe inhibition of T cell responses. The absence of sialic acid residues abolishes these inhibitory activities, showing a key role of sialylated N-glycans in inducing the IgM-mediated immune suppression.

2014-11-11 | E-MTAB-3030 | biostudies-arrayexpress

Transcriptomic profile of Sialyl-Lewis X (CD15s)+ basophils and Sialyl Lewis X- basophils

Project description:Sialyl Lewis X (sLexX or CD15s) is a tetra-saccharide N-acetylneuraminic acid, galactose, fucose and N-acetylglucosamine (NeuAcα2,3Galβ1,4(Fucα1,3)-GlcNAc). CD15s is present on various leukocytes usually as O-linked glycans bound to surface molecules such as PSGL-1, CD43 and CD44. On leukocytes, CD15s plays crucial role in extravasation through the interaction with E-selection expressed ion surface of the activated epithelial blood vessels. We find in approximately 10% of individuals basophils lack CD15s on their surface and these basophils fail to roll on E-selectin coated surfaces. In humans, six α(1,3)-fucosyltransferases (FT), α(1,3)-FTs: FT3/FT4/FT5/FT6/FT7/FT9) catalyses the transfer of fucose to the terminal lactosaminyl glycans giving rise to Lewis X (CD15) and/or sialyl Lewis X (CD15s). To identify which enzyme(s), we perform RNA-sequencing of CD15shigh(CD15s Pos) and CD15slow (CD15s Neg) basophils from 6 individuals.

2021-06-15 | GSE122281 | GEO

Sialylated glycans promote growth in gnotobiotic models

Project description:Sialylated milk glycans promote growth in gnotobiotic mice and pigs with a stunted Malawian infant gut microbiota

| PRJEB9488 | ENA

Growth of B. thetaiotaomicron on purified host mucosal glycans and glycan fragments

Project description:Bacteroides thetaiotaomicron was grown and transcriptionally profiled on a number of different host mucosal glycans and their component mono- and disaccharides.

2008-11-21 | GSE11962 | GEO

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