Gene microarray experiments on RNA samples isolated from murine B cells
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus musculus Homo sapiens
PROVIDER: GSE26480 | GEO | 2011/01/07
SECONDARY ACCESSION(S): PRJNA136517
REPOSITORIES: GEO
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Project description:This laboratory studies 1) enzymes which are involved in selectin ligand formation in leukocytes, 2) how expression of those enzymes is controlled, and 3) basic cell biology of FucT-VII. B cells vs. plasma cells. Study of primary IgG plasma cells. Plasma cells constitute the final stage of B-cell differentiation, and their migration from the lymph nodes and spleen to bone marrow is thought to be an important step in the maintenance of a long-term antibody response to a pathogen. Dr. Kansas has purified primary IgG plasma cells from E-selectin/P-selectin double knockout mice by a negative selection technique and compared their gene expression profile to that of murine B cells using Affymetrix murine oligonucleotide microarray analysis. Highly distinct gene expression profiles were revealed (see Underhill GH et al, Blood. 2003;101:4013-4021). As a follow-up to that experiment, the same probes (in triplicate) derived from primary murine IgG plasma cells and normal B cells were used to probe Glyco-gene Chips. As a follow-up to that experiment, the same probes (in triplicate) derived from primary murine IgG plasma cells and normal B cells were used to probe Glyco-gene Chips.
2011-01-07 | E-GEOD-26480 | biostudies-arrayexpress
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.
2011-10-03 | GSE32556 | GEO
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
Project description:This laboratory works on selectins and their carbohydrate ligands in lymphocyte homing and inflammation. The lab also studies heparin-degrading endosulfatases and their roles in regulating the interactions of growth factors and morphogens with proteoglycans. Gene expression profiles from null and doubly null mice for 2 sulfotransferases (HED-GlcNAc6ST and GlcNAc6ST-1) will be compared to WT mice to find impact on L-selectin ligand formation in lymph nodes. RNA was isolated from the lymph nodes of the 2 types of null mice, 1 type of double null mice, and 1 of wild type mice. 3 duplicates of each strain were analyzed by gene expression profiling. We have null mice and doubly null in two sulfotransferases (HEC-GlcNAc6T and GlcNAc6ST-1) involved in L-selectin ligand formation in lymph nodes. Our functional analysis reveal partial loss (50 and 25%)of lymph node L-selectin ligands in the singly null mice and an 80%loss in the double null mice. We want to identify candidates for the protein scaffolds and underlying posttranslational modifications responsible for the residual L-selectin ligand activity in these mice. We will perform a glycochip analysis with mRNA isolated from lymph nodes of the 3 types of null mice and wild-type mice, all on the C57BL/6 background. Three duplicates from each strain are analyzed, requiring a total of 12 chips. The mice are 8-10 weeks of age and all female.
2011-02-04 | E-GEOD-27056 | biostudies-arrayexpress
Project description:Tumor cell extravasation takes place, similar to leukocytes, in a multi-step process by which cells emigrate from the blood stream through the vascular endothelium into the tissue. Selectins are regarded as the most important molecules for the first capturing step of the cascade. Whether all tumor cells employ selectin dependent adhesion to metastasize or there is an alternative mechanism still is an open question. Comparative transcriptomic analysis of OVCAR8 and SKOV3 cultured cells and revealed that one third of genes encoding proteins involved in the selectin dependent leukocyte like adhesion cascade show lower expression levels in OVCAR8 cultured cells in comparison to that in SKOV3 cultured cells. In contrast to SKOV3 cells, c-Fos overexpression in OVCAR8 cells does not significantly influence expression of the leukocyte like adhesion cascade genes leaving them at similar levels as in control OVCAR8 cells. The intraperitoneal xenograft model of OVCAR8 cells demonstrated that aggressiveness of OVCAR8-derived tumors is not dependent of c-Fos expression level and comparable with that for SKOV3 control tumors. Based on transcriptome array data we analyzed in details expression of genes encoding proteins involved in the leukocyte like adhesion cascade, E- and P-selectin ligands, as well as glycosylation enzymes in both type of tumors. Our results suggest that selectin-dependent mechanism is not the only for adhesion of OVCAR8 ovarian cancer cells. There is at least one additional mechanism of extravasation that does not rely on selectins.
2017-03-31 | GSE97226 | GEO
Project description:This laboratory studies the structures, biosynthesis, and functions of mucin glycans The low-(<33) and high-(>80)passage human prostate LNCaP cells developed by Dr. Ming-Fong Lin (Lin et al, Urology 166:1943, 2001) have been considered as the best in vitro model that reflects the progression of prostate cancer from androgen-dependent to androgen-independent stage (Parajuli et al, Cancer Res. 61:8227, 2001; Demeade et al, The Prostate 54:249, 2003; Unni et al, Cancer Res. 64:7156, 2004). The high-passage cells are metastatic while the low-passage cells are not. RT-PCR analysis of these two cell clones for 25 glycogenes among 8 different groups of glycogenes showed that only high-passage cells expressed two (isozymes 1 and 5) of the five known GlcNAc6ST genes. In addition, P-selectin blot analysis showed qualitative and quantitative differences between these clones. They included 3 higher intensity bands and 3 lower intensity bands in high-passage cells as compared to low-passage cells. The difference in P-selectin-specific glycoconjugates between low and high-passage LNCaP cells should help identify the glycans that distinguish high- from low-passage LNCaP cells.
2011-04-29 | GSE28927 | GEO
Project description:Background. The regenerative and immunomodulatory properties of human mesenchymal stem cells (hMSC) have raised great hope for their use in cell therapy. However, when intravenously infused, hMSCs fail to reach sites of tissue injury. Fucose addition in α-(1,3)-linkage to terminal sialyllactosamines on CD44 creates the molecule known as hematopoietic cell E-/L-selectin ligand (HCELL), programming hMSC binding to E-selectin that is expressed on microvascular endothelial cells of bone marrow (BM), skin, and at all sites of inflammation. Here we describe how this modification on BM-derived hMSC (BM-hMSCs) can be adapted to good manufacturing practice (GMP) standards. Methods. BM-hMSC were expanded using xenogenic-free media and exofucosylated using α-(1,3)-fucosyltransferases VI (FTVI) or VII (FTVII). Enforced fucosylation converted CD44 into HCELL, and HCELL formation was assessed by western blot, flow cytometry, and cell binding assays. Untreated (unfucosylated), buffer-treated and exofucosylated BM-hMSC were each characterized by immunophenotype , differentiation potential and cell viability and fucosylation stability were assessed at room temperature and at 4°C. Safety was assessed by microbiological testing, karyotype, and c-Myc mRNA expression, and potential effects on genetic reprogramming and in cell signaling were analyzed by gene expression microarrays and receptor tyrosine kinase (RTK) phosphorylation arrays. Results. Our protocol efficiently generates HCELL on clinical-scale batches of BM-hMSC. Exofucosylation yields stable HCELL expression for 48 hours at 4°C. Exofucosylation preserved full cell viability and identity, without changing gene expression or RTK phosphorylation. Discussion. The described exofucosylation protocol using xenogenic-free reagents enforces HCELL expression on hMSC endowing potent E-selectin binding without affecting cell viability or native phenotype. This described protocol is readily scalable for GMP-compliant clinical production.
2018-09-05 | GSE90131 | GEO
Project description:Dr. Paulson's group investigates the roles of carbohydrate-binding proteins involved in immune regulation and human disease, through their interaction with carbohydrate groups expressed on cell surface glycoconjugates. This laboratory is currently focused on how sialoside ligands modulate Siglec functions and on understanding the molecular basis of glycosylation changes following differentiation and activation of leukocytes. Three RNA preparations from IL-5 transgenic C567/B1 adult mouse eosinophils were sent to Microarray Core (E). The RNA was amplified, labeled, and hybridized to the GLYCOv3 microarrays. Data was analyzed to determine glycosyltransferase expression on mouse eosinophils, with specific interest in a cis-ligand of Siglec-F on eosinophils
2011-04-29 | E-GEOD-28923 | biostudies-arrayexpress
Project description:How disseminated tumor cells (DTCs) engage specific stromal components in distant organs for survival and outgrowth is a critical but poorly understood step of the metastatic cascade. Previous studies have demonstrated the importance of the epithelial-mesenchymal transition (EMT) in promoting the cancer stem cell properties needed for metastasis initiation, while the reverse process of mesenchymal-epithelial transition (MET) is required for metastatic outgrowth. Here we report that this paradoxical requirement for simultaneous induction of both MET and cancer stem cell traits in DTCs is provided by bone vascular niche E-selectin. Using cell surface alkoxyamine-biotinylation and label-free LC-MS/MS, Glg1 was identified as a top candidate Fut3/Fut6-dependent E-selectin ligand. We functionally validated their involvement in the formation of bone metastasis. These findings provide unique insights into the functional role of E-selectin as a component of the vascular niche criticalfor metastatic colonization in bone.
2019-11-13 | PXD012942 | Pride
Project description:This laboratory studies the structures, biosynthesis, and functions of mucin glycans The low-(<33) and high-(>80)passage human prostate LNCaP cells developed by Dr. Ming-Fong Lin (Lin et al, Urology 166:1943, 2001) have been considered as the best in vitro model that reflects the progression of prostate cancer from androgen-dependent to androgen-independent stage (Parajuli et al, Cancer Res. 61:8227, 2001; Demeade et al, The Prostate 54:249, 2003; Unni et al, Cancer Res. 64:7156, 2004). The high-passage cells are metastatic while the low-passage cells are not. RT-PCR analysis of these two cell clones for 25 glycogenes among 8 different groups of glycogenes showed that only high-passage cells expressed two (isozymes 1 and 5) of the five known GlcNAc6ST genes. In addition, P-selectin blot analysis showed qualitative and quantitative differences between these clones. They included 3 higher intensity bands and 3 lower intensity bands in high-passage cells as compared to low-passage cells. The difference in P-selectin-specific glycoconjugates between low and high-passage LNCaP cells should help identify the glycans that distinguish high- from low-passage LNCaP cells. RNA preparations from low-, intermediate-, and high-passage human prostate LNCaP cells were sent to Microarray Core (E). *Concurrent analysis of glycan profiles in these cells was performed in Core C*. Three replicate samples from each condition were used in the study. The RNA was amplified, labeled, and hybridized to the GLYCOv3 microarrays. Data was analyzed to identify the glycogenes, glycans, and/or carbohydrate binding proteins responsible for the difference in metastatic property between low- and high-passage LNCaP cells. The intermediate-passage cells were used for identifying changes in glycogene, glycan, and/or carbohydrate binding protein expression that may be responsible for the earliest change of the metastatic property.
2011-04-29 | E-GEOD-28927 | biostudies-arrayexpress