To better understand the molecular basis for modulation of glycosaminoglycans (GAGs) and proteoglycans post-UV exposure
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE32552 | GEO | 2011/10/03
SECONDARY ACCESSION(S): PRJNA147017
REPOSITORIES: GEO
Similar Datasets
Project description:UV irradiation is a major environmental effector of skin damage and aging. Elevated levels of glycosaminoglycans (GAGs), as measured by Hale’s stain, are seen following cutaneous photodamage. Preliminary data from our lab indicates that this is a complex response involving differential regulation of both GAGs and proteoglycans. Recently, different GAG species have been shown to have distinct effects on the recruitment and activation of immune cells and stimulation of cytokine production (Taylor and Gallo, FASEB, 2006; 20: 9-22). We speculate that the elevated GAGs and proteoglycans observed after ultraviolet B (UVB) irradiation are involved in the inflammatory and healing responses to photodamage. Chondroitin sulfate synthases (CSSs) are not increased by UVB in mice or in cultured human fibroblasts. To determine whether genomic upregulation of CSSs is responsible for the post-UVB CS increase, we measured the dermal expression of CSS1 and CSS3 mRNA in C57Bl6 mice after 5 days of UV-B exposure. Irradiation caused no change in either CSS1 or CSS3 mRNA expression. We also studied CSS RNA expression in cultured human fibroblasts. We compared control cells to cells treated with 30 mJ/cm2 UVB, cells treated with 1 ng/mL IL-1α, and cells co-stimulated with UVB and IL-1α. In vivo, UV-B induces IL-1α production by keratinocytes and inflammatory cells, and this IL-1α interacts with fibroblasts. Co-stimulation with IL-1α + UVB induces TNF-α production by the fibroblasts, mirroring the in vivo interaction. CSS1 mRNA was suppressed 60% and CSS3 mRNA expression dropped 87% relative to sham at 24 hours post-treatment (p<0.001, Dunnet q’). Since CS is not upregulated by its synthases, we postulated an alternative mode of induction whereby one or more CS proteoglycans are transcriptionally increased and bind more CS in the dermis. We used the N-13 goat monoclonal anti-serglycin antibody to visualize changes in cutaneous serglycin content following acute UV-B exposure. Serglycin is one example of CS-binding dermal proteoglycans that is induced by UVB, and there are likely others. Diffuse upper-dermal serglycin staining, like upper-dermal CS, was induced continuously at 24, 48, and 72 hours after irradiation. Serglycin-expressing inflammatory cells are recruited to the dermis following irradiation, peaking at 48 hours post-exposure. A statistically significant 1.70 fold increase in serglycin mRNA was measured in cultured human fibroblasts 6 hours after co-stimulation with UVB and IL-1α. Realtime PCR also revealed a significant 2.04 fold upregulation at 24 hours after co-stimulation, and serglycin was increased 4.63 times 6 hours after Il-1α treatment alone.
2011-10-02 | E-GEOD-32552 | biostudies-arrayexpress
Project description:Cutaneous lupus erythematosus (CLE) is an autoimmune disease that localizes to the skin and is known to contain elevated glycosaminoglycans (GAGs) on Hale’s stain of skin biopsy specimens. Recently, different GAG species have been shown to have distinct effects on the recruitment and activation of immune cells and stimulation of cytokine production (Taylor and Gallo, FASEB, 2006; 20: 9-22). Thus, we speculate that the elevated GAGs observed in CLE play a role in the local inflammatory process that produces skin lesions in these patients.
2011-06-14 | GSE29938 | GEO
Project description:Cutaneous lupus erythematosus (CLE) is an autoimmune disease that localizes to the skin and is known to contain elevated glycosaminoglycans (GAGs) on Hale’s stain of skin biopsy specimens. Recently, different GAG species have been shown to have distinct effects on the recruitment and activation of immune cells and stimulation of cytokine production (Taylor and Gallo, FASEB, 2006; 20: 9-22). Thus, we speculate that the elevated GAGs observed in CLE play a role in the local inflammatory process that produces skin lesions in these patients. In order to further investigate a molecular basis for the elevated expression of these GAGs in CLE skin lesions, we would like to determine the gene expression profiles of GAG synthesis, degradation, and modifier genes in lesional and non-lesional skin samples from CLE patients and compare to those from healthy controls. A microarray approach will give us a broader understanding of the genetic regulation of the expression of various GAG species in CLE skin. We will then be able to target future quantitative gene expression experiments by real-time RT-PCR to the genes that are shown to be involved in CLE. In order to accomplish our goal, we would like to examine the GAG gene expression profiles of DLE, TLE, and SCLE subtypes due to the differences in CS and HA staining that we found among these subtypes. Since HA and CS are elevated in DLE and HA in TLE, but not in SCLE, the SCLE samples will also serve as an internal control. We would like to examine both lesional and non-lesional skin biopsies to determine if CLE skin prior to developing a lesion is different at the genetic level from healthy control skin and how it changes once a lesion does develop. We will separate the dermis from the epidermis of the skin biopsies and extract RNA just from the dermis to enrich for dermal fibroblast RNA. We aim to submit four patient biopsies per subtype as well as four samples from healthy control skin for comparison. This number is necessary in order to account for the biologic variability among different patients. We would submit more samples per subtype but are limited by availability of patients in clinic. Thus, we will have a total of 28 samples to submit for microarray. This study design will allow us to analyze the GAG gene expression profiles among different CLE subtypes and enable us to identify which GAc
2011-06-14 | E-GEOD-29938 | biostudies-arrayexpress
Project description:To better understand the molecular basis for modulation of glycosaminoglycans (GAGs) and proteoglycans post-UV exposure
| PRJNA147017 | ENA
Project description:Breast cancer mortality results from incurable recurrent tumors, putatively seeded by dormant, therapy-refractory residual tumor cells (RTCs). Understanding the mechanisms enabling RTC survival is therefore essential for improving patient outcomes. We derived a dormancy-associated RTC signature that mirrors the transcriptional response to neoadjuvant chemotherapy in patients and is enriched for extracellular matrix-related pathways. In vivo CRISPR-Cas9 screening of dormancy-associated candidate genes identified the galactosyltransferase B3GALT6 as a functional regulator of RTC fitness. B3GALT6 is required for the linkage of glycosaminoglycans (GAGs) to proteins to generate proteoglycans and its germline loss-of-function causes skeletal dysplasias. We determined that B3GALT6-mediated biosynthesis of heparan sulfate GAGs predicts poor patient outcomes, promotes tumor recurrence by enhancing dormant RTC survival in multiple contexts, and does so via a B3GALT6-heparan sulfate/HS6ST1-heparan 6-O-sulfation/FGF1-FGFR2 signaling axis. These findings implicate B3GALT6 in cancer and suggest targeting of FGFR2 signaling as a novel approach to eradicate dormant RTCs, thereby preventing recurrence.
2024-01-01 | GSE241252 | GEO
Project description:Glioblastoma (GBM) is the most common and malignant primary brain tumor. The extracellular matrix (ECM), also known as the matrisome, helps determine glioma invasion, adhesion, and growth. Little attention, however, has been paid to glycosylation of the ECM components that constitute the majority of glycosylated protein mass and presumed biological properties. To acquire a comprehensive understanding of the biological functions of the matrisome and its components, including proteoglycans and glycosaminoglycans (GAGs), in GBM tumorigenesis, and to identify potential biomarker candidates, we studied the alterations of GAGs, including heparan sulfate (HS) and chondroitin sulfate (CS), the core proteins of proteoglycans, and other glycosylated matrisomal proteins in GBM subtypes vs. control human brain tissue samples. We scrutinized the proteomics data to acquire in-depth site-specific glycoproteomic profiles of the GBM subtypes that will assist in identifying specific glycosylation changes in GBM. We observed an increase in CS 6-O sulfation and a decrease in HS 6-O sulfation, accompanied by an increase in unsulfated CS and HS disaccharides in GBM vs. control samples. Several core matrisome proteins, including proteoglycans (decorin, biglycan, agrin, prolargin, glypican-1, CSPG4), tenascin, fibronectin, hyaluronan link protein 1 and 2, laminins, and collagens, were differentially regulated in GBM vs. controls. Interestingly, a higher degree of collagen hydroxyprolination was also observed for GBM vs. controls. Further, two proteoglycans, CSPG4 and agrin, were significantly lower, about 6–fold for IDH-mutant compared to the WT GBM samples. Differential regulation of O-glycopeptides for proteoglycans, including brevican, neurocan, and versican, was observed for GBM subtypes vs. controls. Moreover, an increase in levels of glycosyltransferase and glycosidase enzymes was observed for GBM when compared to control samples. We also report distinct protein, peptide, and glycopeptide features for GBM subtypes comparisons. Taken together, our study informs understanding of the alterations to key matrisomal molecules that occur during GBM development.
2022-04-04 | PXD028931 | Pride
Project description:We have prepared tissue engineered self assembled dermal equivalents with high similarity to the human dermis (skin). We have manipulated the culture conditions to prepare dermal equivalents with varying amounts of hyaluronic acid and potentially other glycosaminoglycans (GAGs). We are interested in using array technology to determine which genes responsible for carbohydrate and GAG (and if available protein/glycoprotein) synthesis are being either up-or down-regulated in these constructs as compared to a Normal base construct.
2011-10-02 | E-GEOD-32555 | biostudies-arrayexpress
Project description:Here we report a simple and versatile approach for domain mapping of complex mixtures of glycosaminoglycans (GAGs), GAGDoMa. The approach is based on orthogonal enzymatic depolymerization of the GAGs, generating internal oligosaccharide, non-reducing end, and linkage region GAG domains, nanoflow reversed-phase dibutylamine ion-pairing chromatography and negative mode higher-energy collision dissociation (HCD) MS/MS. GAGDoMa provides a detailed insight into the GAGome, and will be an important tool for the understanding of GAGs in cellular physiology and pathology.
2020-02-18 | PXD014504 | Pride
Project description:Advancement in mass spectrometry has revolutionised the field of proteomics. However, there remains a gap in the analysis of protein post-translational modifications (PTMs), particularly for glycosylation. Glycosylation, the most common form of PTM, is involved in most biological processes; thus, analysis of glycans along with proteins is crucial to answering important biologically relevant questions. Of particular interest is the brain extracellular matrix (ECM), which has been called the “final frontier” in neuroscience that consists of highly glycosylated proteins. Among these, proteoglycans (PGs) contain large glycan structures called glycosaminoglycans (GAGs) and form crucial ECM components, including perineuronal nets (PNNs), shown to be altered in neuropsychiatric diseases. Thus, there is a growing need for high-throughput methods that combine GAG (glycomics) and PGs (proteomics) analysis to unravel the complete biological picture. The protocol presented here integrates glycomics and proteomics to analyze multiple classes of biomolecules.
2020-04-30 | PXD017513 | Pride
Project description:We have developed a strategy for the detailed structural characterization of complex proteoglycan-derived glycosaminoglycans. Chondroitin/dermatan sulfate isolated from rat INS-1 832/13 insulinoma cells known to produce primarily one proteoglycan was used to evaluate and demonstrate the efficacy of the strategy.
2021-03-18 | PXD023018 | Pride