chondroitin-sulfates and Mucopolysaccharidosis-I

Topics: beta-Galactosidase; Cells, Cultured; Child, Preschool; Chondroitin Sulfates; Female; Fucose; Gangliosidoses; Genetic Carrier Screening; Hexosyltransferases; Humans; Iduronidase; Liver; Lysosomes; Mannose; Mucolipidoses; Mucopolysaccharidoses; Mucopolysaccharidosis I; Mucopolysaccharidosis II; Mucopolysaccharidosis III; Mucopolysaccharidosis IV; Mucopolysaccharidosis VI

We compared substrate reduction in patients with lysosomal storage disorder treated with hematopoietic stem cell transplant and found that it was significantly reduced compared with patients treated with pharmacological enzyme replacement therapy. These data might support the wider application of hematopoietic stem cell transplant in the treatment of lysosomal storage disorders.

Topics: Biomarkers; Chondroitin Sulfates; Dermatan Sulfate; Enzyme Therapy; Enzymes; Hematopoietic Stem Cell Transplantation; Humans; Mucopolysaccharidosis I; Mucopolysaccharidosis II; Mucopolysaccharidosis VI

Mucopolysaccharidoses (MPSs) are complex lysosomal storage disorders that result in the accumulation of glycosaminoglycans (GAGs) in urine, blood, and tissues. Lysosomal enzymes responsible for GAG degradation are defective in MPSs. GAGs including chondroitin sulfate (CS), dermatan sulfate (DS), heparan sulfate (HS), and keratan sulfate (KS) are disease-specific biomarkers for MPSs. This article describes a stable isotope dilution-tandem mass spectrometric method for quantifying CS, DS, and HS in urine samples. The GAGs are methanolyzed to uronic or iduronic acid-N-acetylhexosamine or iduronic acid-N-sulfo-glucosamine dimers and mixed with internal standards derived from deuteriomethanolysis of GAG standards. Specific dimers derived from HS, DS, and CS are separated by ultra-performance liquid chromatography (UPLC) and analyzed by electrospray ionization tandem mass spectrometry (MS/MS) using selected reaction monitoring for each targeted GAG product and its corresponding internal standard. This UPLC-MS/MS GAG assay is useful for identifying patients with MPS types I, II, III, VI, and VII. © 2023 Wiley Periodicals LLC. Basic Protocol: Urinary GAG analysis by ESI-MS/MS Support Protocol 1: Prepare calibration samples Support Protocol 2: Preparation of stable isotope-labeled internal standards Support Protocol 3: Preparation of quality controls for GAG analysis in urine Support Protocol 4: Optimization of the methanolysis time Support Protocol 5: Measurement of the concentration of methanolic HCl.

Topics: Chondroitin Sulfates; Chromatography, Liquid; Dermatan Sulfate; Glycosaminoglycans; Heparitin Sulfate; Humans; Iduronic Acid; Isotopes; Mucopolysaccharidoses; Mucopolysaccharidosis I; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry

New therapies for the treatment of mucopolysaccharidoses that target the brain, including intrathecal enzyme replacement, are being explored. Quantitative analysis of the glycosaminoglycans (GAGs) that accumulate in these disorders is required to assess the disease burden and monitor the effect of therapy in affected patients. Because current methods lack the required limit of quantification and specificity to analyze GAGs in small volumes of cerebrospinal fluid (CSF), we developed a method based on ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS).. Samples of CSF (25 μL) were evaporated to dryness and subjected to methanolysis. The GAGs were degraded to uronic acid-N-acetylhexosamine dimers and mixed with internal standards derived from deuteriomethanolysis of GAG standards. Specific dimers derived from heparan, dermatan and chondroitin sulfates (HS, DS and CS) were separated by UPLC and analyzed by electrospray ionization MS/MS using selected reaction monitoring for each targeted GAG product and its corresponding internal standard.. CSF from control pediatric subjects (n = 22) contained <0.38 mg/L HS, 0.26 mg/L DS, and 2.8 mg/L CS, whereas CSF from patients with Hurler syndrome (n = 7) contained concentrations of DS and HS that were at least 6-fold greater than the upper control limits. These concentrations were reduced by 17.5% to 82.5% after allogeneic transplantation and treatment with intrathecal and intravenous enzyme replacement therapy.. The method described here has potential value in monitoring patients with mucopolysaccharidoses receiving treatment targeted to the brain.

Topics: Biomarkers; Calibration; Child; Chondroitin Sulfates; Chromatography, High Pressure Liquid; Dermatan Sulfate; Deuterium; Dimerization; Enzyme Replacement Therapy; Hematopoietic Stem Cell Transplantation; Heparitin Sulfate; Hexosamines; Humans; Indicator Dilution Techniques; Injections, Intravenous; Injections, Spinal; Mucopolysaccharidosis I; Reference Standards; Reference Values; Tandem Mass Spectrometry; Uronic Acids

Hurler Syndrome is corrected by allogeneic BMT by the action of donor enzyme on recipient tissue. In this paper, we describe monitoring of 39 patients transplanted in two centres to determine donor chimerism, enzyme level and residual substrate - expressed as dermatan sulphate to chondroitin sulphate ratio. We show that in fully engrafted recipients, the enzyme level, expressed as mumol/g total protein/h, post-transplant is 24.2 from an unrelated donor and 10.2 from a heterozygote family donor (P<0.0001). There is a tight relationship between mean post-transplant enzyme level and residual substrate - Spearman's rank correlation coefficient (Rho) was -0.76 and -0.80 at 12 and 24 months, respectively (P<0.0001). We propose that these differences affect patient outcome. As unrelated donor transplant outcomes improve and especially given the higher levels of donor cell engraftment following cord transplants, our data might influence donor selection where only heterozygote-matched family members are available.

Topics: Chimerism; Chondroitin Sulfates; Cord Blood Stem Cell Transplantation; Dermatan Sulfate; Glycosaminoglycans; Hematopoietic Stem Cell Transplantation; Heterozygote; Histocompatibility Testing; Humans; Iduronidase; Mucopolysaccharidosis I; Transplantation, Homologous; Treatment Outcome

The phenotypic resemblance of patients with Costello syndrome and Hurler disease has been linked to impaired formation of elastic fibers that coincides with elevated cellular proliferation. Impaired elastogenesis in these diseases associates with respective abnormal accumulation of chondroitin sulfate and dermatan sulfate proteoglycans that induce cell surface shedding of elastin-binding protein (EBP) normally required for intracellular chaperoning of tropoelastin and its assembly into elastic fibers. A variant of the chondroitin sulfate proteoglycan versican, V3, which lacks chondroitin sulfate, has recently been shown to stimulate elastic fiber assembly and decrease proliferation when expressed by retroviral transduction in arterial smooth muscle cells. However, the mechanism(s) by which V3 influences this phenotype is not known. We now demonstrate that transduction of skin fibroblasts from Costello syndrome and Hurler disease patients with cDNA to versican V3 completely reverses impaired elastogenesis and restores normal proliferation of these cells. This phenotypic reversal is accompanied by loss of chondroitin sulfate from the cell surface and increased levels of EBP. Versican V3 transduction of skin fibroblasts from GM(1)-gangliosidosis patients, which lack EBP, failed to restore impaired elastogenesis. These results suggest that induction of elastic fiber production by gene transfer of versican V3 in skin fibroblasts is mediated by rescue of the tropoelastin chaperone, EBP.

Topics: Animals; Blotting, Northern; Cell Division; Cells, Cultured; Child, Preschool; Chondroitin Sulfate Proteoglycans; Chondroitin Sulfates; Elastic Tissue; Female; Fibroblasts; Gangliosidosis, GM1; Humans; Immunohistochemistry; Infant; Lectins, C-Type; Male; Mucopolysaccharidosis I; Receptors, Cell Surface; Retroviridae; RNA, Messenger; Transduction, Genetic; Up-Regulation; Versicans

Vitamin A decreased the urinary excretion of total mucopolysaccharides in a patient with Hurler-Scheie compound (type IH-S mucopolysaccharidosis). Vitamin A was administered orally in daily doses of 1,000 to 2,000 IU/kg body weight for 10 years. Adverse clinical responses such as irritability, bone pain, dizziness, vomiting and diarrhea appeared in the patient and were controlled by reduction of the dose administered. No clinical improvement was observed, although it is possible that the clinical course of the disease may have been retarded.

Topics: Adolescent; Chondroitin Sulfates; Dermatan Sulfate; Dose-Response Relationship, Drug; Female; Glycosaminoglycans; Heparitin Sulfate; Humans; Mucopolysaccharidosis I; Syndrome; Vitamin A

The effect of two years of periodic transfusions at 2--4 month intervals of mixed leukocytes in a patient with Type I mucopolysaccharidosis is described. There was an increase of urinary chondroitin sulfate B after the first transfusion followed by a progressive decrease to 35% of the initial daily excretion. Chondroitin sulfate AC, which was almost absent prior to the transfusions, was excreted in significant amounts after the 4th transfusion. There was a decrease of hepatosplenomegaly, kyphosis and gibbus and no changes in the other signs of the disease. Regarding stature, mental development and social behavior, the patient is developing thus far as a normal child. The early diagnosis of the disease and the immediate transfusions undoubtedly have been contributing factors to the restoration and/or maintenance of the child in an almost normal condition.

Topics: Blood Transfusion; Chondroitin Sulfates; Female; Glycosaminoglycans; Humans; Infant; Leukocyte Transfusion; Mucopolysaccharidosis I

Crude glycosaminoglycan (GAG) fraction was directly precipitated with cetylpyridinium chloride without prior dialysis of urine of orthopedic patients. The crude GAG fraction was then fractionated with trichloroacetic acid (TCA). The TCA-insoluble peptide-bound GAG fraction thus obtained was treated with alkali to eliminate the peptide moiety for enzymatic analysis. The GAG compositions of this fraction and the TCA-soluble fraction were determined by digestion with mucopolysaccharidases (chondroitinase AC, chondroitinase B, chondroitinase C, heparitinase and Streptomyces hyaluronidase). When the amount of the crude GAG fraction was small, no significant amount of the TCA-insoluble peptide-bound GAG fraction was obtained. The GAG composition of this case was also determined by the same procedures after direct alkali-treatment of the crude GAG fraction. The data indicated that the proportion of the TCA-insoluble peptide-bound GAG fraction was very small. The alkali-treated TCA-insoluble peptide-bound GAG fraction contained a larger proportion of heparan sulfate than the TCA-soluble GAG fraction. It was clearly demonstrated that the patients with Werner's syndrome and mucopolysaccharidosis I-S (Scheie) excreted large amounts of hyaluronic acid and dermatan sulfate respectively, into urines. It was indicated in most cases that major urinary GAG were chondroitin 4-sulfate, chondroitin 6-sulfate plus chondroitin and heparan sulfate, while minor ones were dermatan sulfate and hyaluronic acid. In addition, the data suggested a wide range of the degree of desulfation or urinary GAG, and the presence of significant amounts of keratan sulfate plus acidic glycopeptides in the urinary GAG fractions. The present data provided more precise information on urinary GAG from orthopedic patients than those reported previously.

Topics: Adolescent; Adult; Aged; Bone Diseases; Chemical Fractionation; Chondroitin Sulfates; Dermatan Sulfate; Female; Glucuronidase; Glycosaminoglycans; Heparitin Sulfate; Humans; Hyaluronic Acid; Lyases; Male; Middle Aged; Mucopolysaccharidosis I; Werner Syndrome

The mucopolysaccharidoses are a group of genetic diseases characterized by storage of incompletely degraded glycosaminoglycans. Such storage causes marked distortion of many tissues with consequent severe somatic changes and mental retardation. Storage of glycosaminoglycans results from markedly diminished activity of specific hydrolases requisite for the normal degradation of glycosaminoglycans. The specific enzymic defects have been identified in nine different diseases. In some cases evidence has been obtained indicating the existence of additional allelic diseases based on the same enzyme. The knowledge obtained from these studies has made prenatal diagnosis possible and has led to the possibility that therapy may be undertaken utilizing enzyme replacement.

Topics: Acetylglucosaminidase; Alleles; Arylsulfatases; Chondroitin Sulfates; Dermatan Sulfate; Glucuronidase; Glycosaminoglycans; Heparitin Sulfate; History, 20th Century; Humans; Iduronidase; Mucopolysaccharidoses; Mucopolysaccharidosis I; Sulfatases

Topics: Animals; Cattle Diseases; Chondroitin Sulfates; Dwarfism; Glycosaminoglycans; Hydrolases; Mucopolysaccharidosis I; Research; Spectrophotometry; Urine