heparitin-sulfate and Glioblastoma

Brain tumors are aggressive and devastating diseases. The most common type of brain tumor, glioblastoma (GBM), is incurable and has one of the worst five-year survival rates of all human cancers. GBMs are invasive and infiltrate healthy brain tissue, which is one main reason they remain fatal despite resection, since cells that have already migrated away lead to rapid regrowth of the tumor. Curative therapy for medulloblastoma (MB), the most common pediatric brain tumor, has improved, but the outcome is still poor for many patients, and treatment causes long-term complications. Recent advances in the classification of pediatric brain tumors reveal distinct subgroups, allowing more targeted therapy for the most aggressive forms, and sparing children with less malignant tumors the side-effects of massive treatment. Heparan sulfate proteoglycans (HSPGs), main components of the neurogenic niche, interact specifically with a large number of physiologically important molecules and vital roles for HS biosynthesis and degradation in neural stem cell differentiation have been presented. HSPGs are composed of a core protein with attached highly charged, sulfated disaccharide chains. The major enzyme that degrades HS is heparanase (HPSE), an important regulator of extracellular matrix (ECM) remodeling which has been suggested to promote the growth and invasion of other types of tumors. This is of clinical interest because GBM are highly invasive and children with metastatic MB at the time of diagnosis exhibit a worse outcome. Here we review the involvement of HS and HPSE in development of the nervous system and some of its most malignant brain tumors, glioblastoma and medulloblastoma.

Topics: Brain Neoplasms; Glioblastoma; Glucuronidase; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; Medulloblastoma

Glioblastoma (GBM) is the most common and malignant primary brain tumor. The extracellular matrix, also known as the matrisome, helps determine glioma invasion, adhesion, and growth. Little attention, however, has been paid to glycosylation of the extracellular matrix 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 (PGs) 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 PGs, and other glycosylated matrisomal proteins in GBM subtypes versus 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 versus control samples. Several core matrisome proteins, including PGs (decorin, biglycan, agrin, prolargin, glypican-1, and chondroitin sulfate proteoglycan 4), tenascin, fibronectin, hyaluronan link protein 1 and 2, laminins, and collagens, were differentially regulated in GBM versus controls. Interestingly, a higher degree of collagen hydroxyprolination was also observed for GBM versus controls. Further, two PGs, chondroitin sulfate proteoglycan 4 and agrin, were significantly lower, about 6-fold for isocitrate dehydrogenase-mutant, compared to the WT GBM samples. Differential regulation of O-glycopeptides for PGs, including brevican, neurocan, and versican, was observed for GBM subtypes versus 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. (Data are available via ProteomeXchange with identifier PXD028931, and the peaks project file is available at Zenodo with DOI 10.5281/zenodo.5911810).

Topics: Agrin; Brain; Brain Neoplasms; Chondroitin Sulfate Proteoglycans; Extracellular Matrix; Extracellular Matrix Proteins; Glioblastoma; Glycosaminoglycans; Heparitin Sulfate; Humans

Signaling from multiple receptor tyrosine kinases (RTK) contributes to therapeutic resistance in glioblastoma (GBM). Heparan sulfate (HS), present on cell surfaces and in the extracellular matrix, regulates cell signaling via several mechanisms. To investigate the role for HS in promoting RTK signaling in GBM, we generated neural progenitor cells deficient for HS by knockout of the essential HS-biosynthetic enzyme

Topics: Animals; Cell Proliferation; Disease Models, Animal; ErbB Receptors; Glioblastoma; Heparitin Sulfate; Humans; Mice; N-Acetylglucosaminyltransferases; Receptor Protein-Tyrosine Kinases; Signal Transduction

Glycosaminoglycans (GAGs) such as heparan sulfate and chondroitin sulfate decorate all mammalian cell surfaces. These mucopolysaccharides act as coreceptors for extracellular ligands, regulating cell signaling, growth, proliferation, and adhesion. In glioblastoma, the most common type of primary malignant brain tumor, dysregulated GAG biosynthesis results in altered chain length, sulfation patterns, and the ratio of contributing monosaccharides. These events contribute to the loss of normal cellular function, initiating and sustaining malignant growth. Disruption of the aberrant cell surface GAGs with small molecule inhibitors of GAG biosynthetic enzymes is a potential therapeutic approach to blocking the rogue signaling and proliferation in glioma, including glioblastoma. Previously, 4-azido-xylose-α-UDP sugar inhibited both xylosyltransferase (XYLT-1) and β-1,4-galactosyltransferase-7 (β-GALT-7)-the first and second enzymes of GAG biosynthesis-when microinjected into a cell. In another study, 4-deoxy-4-fluoro-β-xylosides inhibited β-GALT-7 at 1 mM concentration

Topics: Animals; Cell Line, Tumor; Chondroitin Sulfates; Galactosyltransferases; Glioblastoma; Glycosaminoglycans; Glycosides; Heparitin Sulfate; Humans; Molecular Docking Simulation; Pentosyltransferases; Prodrugs; UDP Xylose-Protein Xylosyltransferase

Adjuvant chemoradiotherapy is a standard treatment option for glioblastoma multiforme (GBM). Despite intensive care, recurrent tumors developed during the first year are fatal for the patients. Possibly contributing to this effect, among other causes, is that therapy induces changes of polysaccharide heparan sulfate (HS) chains in the cancer cells and/or tumor microenvironment. The aim of this study was to perform a comparative analysis of heparanase (HPSE) expression and HS content in different normal and GBM brain tissues. Immunohistochemical analysis revealed a significant decrease of HPSE protein content in the tumor (12-15-fold) and paratumorous (2.5-3-fold) GBM tissues compared with normal brain tissue, both in cellular and extracellular compartments. The relapsed GBM tumors demonstrated significantly higher intertumor and/or intratumor heterogeneity of HPSE and HS content and distribution compared with the matched primary ones (from the same patient) (

Topics: Adult; Aged; Brain; Brain Neoplasms; Cell Line, Tumor; Chemoradiotherapy, Adjuvant; Female; Glioblastoma; Glucuronidase; Heparitin Sulfate; Humans; Male; Middle Aged; Neoplasm Grading; Neoplasm Recurrence, Local

Glycosaminoglycans are major components of brain extracellular matrix (ECM), although heparan sulfate (HS) contribution in brain physiology and carcinogenesis remains underinvestigated. This study examined HS content and distribution in glioblastoma multiforme (GBM) tissues in the context of potential molecular mechanisms underlying its deregulation in brain tumours. Totally, 42 tissue samples and paraffin-embedded tissues for 31 patients with different prognosis were investigated. HS expression was demonstrated in 50-55% of the GBM tumours by immunohistochemistry (IHC), while almost no HS content was detected in the surrounding paratumourous brain tissues. Heterogeneous HS distribution in the HS-positive tumours was more related to the necrosis or glandular-like brain zones rather than glioma cells with high or low Ki-67 index. According the Kaplan-Meier curves, HS accumulation in glioma cells was associated with low relapse-free survival (RS) of the GBM patients (p < 0.05) and was likely to be due to the increased transcriptional activity of HSPG core proteins (syndecan-1, 2-3 fold; glypican-1, 2,5 fold; perlecan/HSPG2, 13-14 fold). Activation of perlecan/HSPG2 expression correlated with the patients' survival according Kaplan-Meier (p = 0.0243) and Cox proportional-hazards regression (HR = 3.1; P(Y) = 0.03) analyses, while up-regulation of syndecan-1 and glypican-1 was not associated with the patients survival. Taken together, the results indicate that increase of HS content and up-regulation of perlecan/HSPG2 expression in glioblastoma tissues contribute to tumour development through the transformation of brain extracellular matrix into tumour microenvironment, and represent negative prognostic factors for glioblastoma progression.

Topics: Brain Neoplasms; Glioblastoma; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; Middle Aged; Recurrence; Survival Analysis; Up-Regulation

Glioblastoma (GBM) is the most common primary malignant brain tumor of adults and confers a poor prognosis due, in part, to diffuse invasion of tumor cells. Heparan sulfate (HS) glycosaminoglycans, present on the cell surface and in the extracellular matrix, regulate cell signaling pathways and cell-microenvironment interactions. In GBM, the expression of HS glycosaminoglycans and the enzymes that regulate their function are altered, but the actual HS content and structure are unknown. However, inhibition of HS glycosaminoglycan function is emerging as a promising therapeutic strategy for some cancers. In this study, we use liquid chromatography-mass spectrometry analysis to demonstrate differences in HS disaccharide content and structure across four patient-derived tumorsphere lines (GBM1, 5, 6, 43) and between two murine tumorsphere lines derived from murine GBM with enrichment of mesenchymal and proneural gene expression (mMES and mPN, respectively) markers. In GBM, the heterogeneous HS content and structure across patient-derived tumorsphere lines suggested diverse functions in the GBM tumor microenvironment. In GBM5 and mPN, elevated expression of sulfatase 2 (SULF2), an extracellular enzyme that alters ligand binding to HS, was associated with low trisulfated HS disaccharides, a substrate of SULF2. In contrast, other primary tumorsphere lines had elevated expression of the HS-modifying enzyme heparanase (HPSE). Using gene editing strategies to inhibit HPSE, a role for HPSE in promoting tumor cell adhesion and invasion was identified. These studies characterize the heterogeneity in HS glycosaminoglycan content and structure across GBM and reveal their role in tumor cell invasion.

Topics: Animals; Brain Neoplasms; Cell Line, Tumor; Chromatography, Liquid; Gene Editing; Glioblastoma; Glucuronidase; Heparitin Sulfate; Humans; Mass Spectrometry; Mice; Neoplasm Invasiveness; Neoplasm Transplantation; Signal Transduction; Sulfatases; Sulfotransferases; Tumor Microenvironment

Heparan sulfate (HS) is an important component of the extracellular matrix and cell surface, which plays a key role in cell-cell and cell-matrix interactions. Functional activity of HS directly depends on its structure, which determined by a complex system of HS biosynthetic enzymes. During malignant transformation, the system can undergo significant changes, but for glioma, HS biosynthesis has not been studied in detail. In this study, we performed a comparative analysis of the HS biosynthetic system in human gliomas of different grades. RT-PCR analysis showed that the overall transcriptional activity of the main HS biosynthesis-involved genes (

Topics: Adult; Biosynthetic Pathways; Brain Neoplasms; Down-Regulation; Female; Glioblastoma; Glioma; Heparitin Sulfate; Humans; Male; Middle Aged; N-Acetylglucosaminyltransferases; Sulfotransferases; Tumor Microenvironment

Glycosaminoglycans (GAGs) are found in intracellular granules, cell surfaces, and extracellular matrices in a spatially and temporally regulated fashion, constituting the environment for cells to interact, migrate, and proliferate. Through binding with a great number of proteins, GAGs regulate many facets of biological processes from embryonic development to normal physiological functions. GAGs have been shown to be involved in pathologic changes and immunological responses including cancer metastasis and inflammation. Past analyses of GAGs have focused on cell lines, body fluids, and relatively large tissue samples. Structures determined from such samples reflect the heterogeneity of the cell types present. To gain an understanding of the roles played by GAG expression during pathogenesis, it is very important to be able to detect and profile GAGs at the histological scale so as to minimize cell heterogeneity to potentially inform diagnosis and prognosis. Heparan sulfate (HS) belongs to one major class of GAGs, characterized by dramatic structural heterogeneity and complexity. To demonstrate feasibility of analysis of HS, 15 μm frozen bovine brain stem, cortex, and cerebellum tissue sections were washed with a series of solvent solutions to remove lipids before applying heparin lyases I, II, and III on the tissue surfaces within 5 mm × 5 mm digestion spots. The digested HS disaccharides were extracted from tissue surfaces and then analyzed by using size exclusion chromatography/mass spectrometry (SEC-MS). The results from bovine brain stem, cortex, and cerebellum demonstrated the reproducibility and reliability of our profiling method. We applied our method to detect HS from human astrocytoma (WHO grade II) and glioblastoma (GBM, WHO grade IV) frozen slides. Higher HS abundances and lower average sulfation level of HS were detected in glioblastoma (GBM, WHO grade IV) slides compared to astrocytoma (WHO grade II) slides.

Topics: Animals; Astrocytoma; Brain Stem; Cattle; Cerebellum; Cerebral Cortex; Chromatography, Gel; Glioblastoma; Heparitin Sulfate; Humans; Neoplasm Grading; Spectrometry, Mass, Electrospray Ionization

Interferon-gamma (IFN-gamma) is a potent immune regulatory cytokine and is, in addition, involved in the induction of antiparasitic effector mechanisms in different cell types. The first step of IFN-gamma action is its binding to a specific receptor. Furthermore, it has been shown that IFN-gamma binds with a great affinity to the heparin-like structure of heparan sulfate, which is localized in basement membranes and on cell surfaces. In this study, we analyze the effect of heparin and heparan sulfate on three different IFN-gamma-mediated activities inducible in human glioblastoma cells (87HG31 and 86HG39). We find firstly that heparin is able to inhibit IFN-gamma-mediated induction of major histocompatibility complex (MHC) class II antigen expression on 87HG31 cells, an effect which can be abrogated by protamine. Secondly, we show that heparin inhibits the IFN-gamma-induced toxoplasmostasis within 86HG39 cells in a dose-dependent fashion, and thirdly that heparin inhibits the IFN-gamma-mediated induction of the tryptophan-degrading enzyme indoleamine 2,3-dioxygenase. In contrast to IFN-gamma-induced effects, the activity of other cytokines, such as interleukin (IL)-1, IL-2 and IL-6, is not influenced by heparin. The possible mechanism of heparin-induced inhibition of IFN-gamma is discussed.

Topics: Animals; Cytokines; Glioblastoma; Heparin; Heparitin Sulfate; HLA-DR Antigens; Humans; Interferon-gamma; Recombinant Proteins; Toxoplasma; Tryptophan Oxygenase; Tumor Cells, Cultured

A total of 14 surgical specimens, including 7 glioblastomas, 3 anaplastic astrocytomas, 2 brains adjacent to glioblastoma and 2 grossly normal brains, were investigated immunohistochemically for the expression of antithrombin III (AT-III), heparan sulfate proteoglycan (HSPG) and thrombomodulin (TM) in the endothelium of microvessels. The immunoreaction to AT-III was of moderate intensity in grossly normal brains, brains adjacent to glioblastoma, and anaplastic astrocytomas, but was only weak in glioblastomas, especially in the capillaries. The immunoreaction to HSPG was constantly intense in the microvessels in all the specimens. Although the immunoreaction to TM was negative or only faint in the microvessels in grossly normal brains, it was moderately to strongly intense in anaplastic astrocytomas and brains adjacent to glioblastoma. The intensity of immunoreaction to TM was variable, from faint to strong in the capillaries, and moderate to strong in larger microvessels in glioblastomas. The present study suggested that the alterations in the expression of those antithrombotic molecules could explain, at least in part, the tendencies for intratumoral hemorrhage as well as intravascular thrombosis in the different areas of malignant gliomas.

Topics: Antithrombin III; Brain; Brain Neoplasms; Endothelium, Vascular; Glioblastoma; Glioma; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; Immunohistochemistry; Microcirculation; Proteoglycans; Reference Values; Thrombomodulin