heparitin-sulfate and Scleroderma--Systemic

The study of the heparanase has long been paid wide attention. Heparanase, an endo-β-D-glucuronidase, is capable of specifically degrading heparan sulfate (HS), one of the excellular matrix (ECM) components. It exerts its enzymatic activity catalyzing the cleavage of the β (1,4)-glycosidic bond between glucuronic acid and glucosamine residue. HS cleavage results in remodelling of the extracellular matrix as well as in regulating the release of many HS-linked molecules such as growth factors, cytokines and enzymes involved in inflammation, wound healing and tumour invasion. Varieties of experiments indicated that heparanase mRNA is overexpressed in human tumors, including breast cancer, gastrointestinal tumors, and esophageal carcinomas. A pro-metastatic and pro-angiogenic role for heparanase has been widely verified and high levels of heparanase correlate with reduced survival of cancer patients. Except protumor function, heparanase also plays a role in inflammation, angiogenesis, placentas and procoagulant activities. Heparanase is found to have many other functions in recent years, since many experiments have been carried out to identify this significant enzyme's new features. These newly found functions are related to the cellular activities such as autophagy and epithelial to mesenchymal transition (EMT). And together with other heparanase functions, autophagy and EMT are verified to be involved in several clinical disorders, for example, renal diseases. Considering that, once inactivated, there are no other enzymes capable of performing the same function, it is apparent that heparanase can be an effective and promising therapy target. This short review aims to establish the currently known function of this enzyme and provide evidence for heparanase targeted therapy.

Topics: Autophagy; Epithelial-Mesenchymal Transition; Glucuronidase; Heparitin Sulfate; Humans; Inflammation; Kidney Diseases; Neoplasms; Prion Diseases; Scleroderma, Systemic

Topics: Animals; Antibodies, Antinuclear; Autoantibodies; Disease Models, Animal; DNA Topoisomerases, Type I; Heparitin Sulfate; Humans; Mice; Mice, Inbred Strains; Receptors, Fc; RNA Polymerase I; Scleroderma, Systemic

Previous studies have demonstrated that the systemic sclerosis (SSc)-associated autoantigen DNA topoisomerase I (topo I) binds specifically to the surface of fibroblasts when released in the extracellular environment and recruits anti-topo I autoantibodies, which subsequently leads to the adhesion and activation of monocytes. This study aimed to characterize the molecular interactions of topo I with fibroblast surfaces in order to elucidate the pathogenic role of topo I/anti-topo I immune complexes (ICs) in SSc.. Topo I directly coupled to fluorochromes was used to follow its binding to fibroblast surfaces by flow cytometry and fluorescence microscopy. Purified IgG from normal subjects or SSc patients was added with topo I to the cells; unfractionated heparin (UFH) and low molecular weight heparin (LMWH) were used to determine their effects on the binding of topo I and topo I/anti-topo I IC to fibroblast surfaces.. Heparan sulfate (HS) proteoglycans on fibroblast surfaces were found to act as coreceptors for topo I binding. The addition of anti-topo I autoantibodies from SSc sera led to the amplification of topo I binding to HS chains. UFH and LMWH were shown to inhibit topo I and topo I/anti-topo I IC binding to HS chains.. This study is the first to show that topo I binds specifically to HS proteoglycans on fibroblast surfaces and that anti-topo I autoantibodies from SSc patients amplify topo I binding to HS chains. The accumulation of topo I on cell surfaces by anti-topo I autoantibodies could contribute to the initiation of an inflammatory cascade stimulating the fibrosis. UFH and LMWH inhibited the binding of topo I/anti-topo I IC to fibroblasts, suggesting a potential therapeutic role in SSc-associated fibrosis.

Topics: Antigen-Antibody Complex; Antigens, Surface; Autoantibodies; Cells, Cultured; Dermis; DNA Topoisomerases, Type I; Fibrinolytic Agents; Fibroblasts; Heparin; Heparitin Sulfate; Humans; Protein Binding; Scleroderma, Systemic

To investigate the contribution of heparan sulfate proteoglycan and Ras/MEK/ERK to the overexpression of profibrotic proteins and the enhanced contractile ability of dermal fibroblasts from patients with systemic sclerosis (SSc; scleroderma).. The effects of the MEK/ERK inhibitor U0126, the heparan sulfate side chain formation inhibitor beta-xyloside, and soluble heparin on the overexpression of profibrotic genes were compared in fibroblasts from lesional skin of patients with diffuse SSc and fibroblasts from healthy control subjects. Identified protein expressions were compared with the contractile abilities of fibroblasts while they resided within a collagen lattice. Forces generated were measured using a culture force monitor.. Inhibiting MEK/ERK with U0126 significantly reduced expression of a cohort of proadhesive and procontractile proteins that normally are overexpressed by scleroderma fibroblasts, including integrin alpha4 and integrin beta1. Antagonizing heparan sulfate side chain formation with beta-xyloside or the addition of soluble heparin prevented ERK activation, in addition to reducing the expression of these proadhesive/contractile proteins. Treatment with either U0126, beta-xyloside, or heparin resulted in a reduction in the overall peak contractile force generated by dermal fibroblasts. Blocking platelet-derived growth factor receptor with Gleevec (imatinib mesylate) reduced overall contractile ability and the elevated syndecan 4 expression and ERK activation in SSc fibroblasts.. The results of this study suggest that heparan sulfate-dependent ERK activation contributes to the enhanced contractile ability demonstrated by dermal fibroblasts from lesional skin of patients with scleroderma. These results are consistent with the notion that the MEK/ERK procontractile pathway is dysregulated in scleroderma dermal fibroblasts. Additionally, the results suggest that antagonizing the MEK/ERK pathway is likely to modulate heparan sulfate proteoglycan activity, which in turn may have a profound effect on the fibrotic response in SSc.

Topics: Cell Movement; Cells, Cultured; Dermis; Extracellular Matrix; Extracellular Signal-Regulated MAP Kinases; Female; Fibroblasts; Fibrosis; Gene Expression; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Humans; MAP Kinase Signaling System; Phenotype; Platelet-Derived Growth Factor; RNA, Messenger; Scleroderma, Systemic; Syndecan-4

We report a case of atrophoderma of Pasini and Pierini. We determined the glycosaminoglycan content in the involved skin. Dermatan sulfate content in the involved skin (1.88 micrograms uronic acid/mg dry skin) was greater than that in the uninvolved skin (1.05 micrograms uronic acid/mg dry skin). No significant differences in hyaluronic acid, chondroitin sulfate or heparan sulfate content between involved and uninvolved skin were observed. These results suggest that abnormal metabolism of dermatan sulfate may be involved in the pathogenesis of atrophoderma; this pattern has been observed in systemic or localized scleroderma.

Topics: Adult; Atrophy; Chondroitin Sulfates; Dermatan Sulfate; Glycosaminoglycans; Heparitin Sulfate; Humans; Hyaluronic Acid; Male; Pigmentation Disorders; Scleroderma, Localized; Scleroderma, Systemic; Skin; Uronic Acids

As in human scleroderma, tight skin (TSK/+) mice develop cutaneous hyperplasia with over-production of extracellular matrix, including collagen and proteoglycans, associated with autoimmunity to a number of autoantigens. The present study investigated the presence of cellular autoimmunity to basement membrane heparan sulfate proteoglycan (HSPG) in diseased TSK/+mice and their nondiseased littermates, pallid mice (+/pa). Lymphocyte proliferative responses to specific HSPG antigens, including intact HSPG, HSPG protein core (P. Core), and the glycosaminoglycan heparan sulfate (HS) were studied. Splenocytes from young TSK and control pallid mice reacted weakly to intact HSPG and HSPG P. Core antigens and did not respond to HS. However, lymphocytes from old TSK mice were reactive with HSPG and P. Core and demonstrated a de novo response to HS. The proliferative response to intact HSPG and HSPG P. Core in TSK mice was T cell dependent, but the response to HS was T cell independent. The T cell-dependent response was mediated by the CD4-positive subset and required the participation of class II major histocompatibility complex molecules. Cellular autoimmunity to HSPG, a critical cell surface and extracellular matrix component, may play a role in the disease suffered by TSK mice. Further studies are necessary to determine the mechanisms which link autoimmunity to HSPG with the pathology seen in TSK mice, particularly the overproduction of extracellular matrix and fibrosis.

Topics: Animals; Cattle; CD4 Antigens; CD8 Antigens; Heparan Sulfate Proteoglycans; Heparitin Sulfate; Histocompatibility Antigens Class II; Immunity, Cellular; Lymphocyte Activation; Lymphocyte Depletion; Lymphocyte Subsets; Mice; Mice, Inbred C57BL; Mice, Mutant Strains; Proteoglycans; Scleroderma, Systemic

Systemic lupus erythematosus (SLE) is an autoimmune disease which involves the basement membranes of blood vessels in multiple organs. An important component of the microvasculature is vascular heparan sulfate proteoglycan (HSPG). In this study, we investigated the presence in SLE and other immune disease sera of autoantibodies to purified vascular HSPG. Our data demonstrate that antibody to HSPG is found primarily in SLE sera, and not in sera from controls or patients with other immune diseases. The titer of antibody to HSPG correlated with complement depletion in SLE sera. Antibody to HSPG was frequently found in high titer in SLE patients with renal and neurologic involvement. These studies indicate that our assay for antibody to vascular HSPG detects a pathologically relevant autoantibody in SLE sera. The implications of our findings for pathogenesis of vascular autoimmunity are discussed, including the relationship of anti-vascular HSPG antibody to anti-DNA and antiphospholipid antibodies.

Topics: Administration, Topical; Anti-Inflammatory Agents; Antibody Specificity; Arthritis, Rheumatoid; Basement Membrane; Blotting, Western; Chondroitin Sulfates; Collagen; Complement System Proteins; Dose-Response Relationship, Immunologic; Enzyme-Linked Immunosorbent Assay; Fibronectins; Heparin; Heparitin Sulfate; Humans; Hyaluronic Acid; Laminin; Lupus Erythematosus, Systemic; Muscle, Smooth, Vascular; Polymyalgia Rheumatica; Proteoglycans; Scleroderma, Systemic; Sjogren's Syndrome

The compositional changes of acidic glycosaminoglycans (AGAG) in the urine of progressive systemic sclerosis (PSS) patients were studied using chondroitinases and heparitinase in appropriate enzyme assays and by electrophoretic characterization. Daily urinary excretion of AGAG in patients with PSS was increased, when compared to normals. The proportion of urinary AGAG in PSS patients, which was undigested by chondroitinase-ABC (most probably representing heparan sulfates (HS)), increased significantly from the normal value. The substance was found to be mainly HS as determined by the electrophoretic pattern, thin-layer chromatographic analysis and by its susceptibility to heparitinase. After digestion of urinary chondroitin sulfate isomers with chondroitinases, the unsaturated disaccharides were mainly separated into 4-sulfated and 6-sulfated disaccharide units by paper chromatography. In all of the patients with PSS, the ratio of the unsaturated 4-sulfated disaccharide to the unsaturated disaccharide was higher than that in normal subjects. These observations indicate an abnormal turnover of AGAG in patients with PSS.

Topics: Adolescent; Adult; Aging; Chondroitin Sulfates; Chondroitinsulfatases; Chromatography, Gel; Chromatography, Paper; Chromatography, Thin Layer; Electrophoresis, Cellulose Acetate; Female; Glycosaminoglycans; Heparitin Sulfate; Humans; Male; Middle Aged; Polysaccharide-Lyases; Scleroderma, Systemic; Uronic Acids