title: A method for measuring disease-specific iduronic acid from the non-reducing end of glycosaminoglycan in mucopolysaccharidosis type II mice.
abstract: Mucopolysaccharidosis type II (MPS II) is an X-linked lysosomal storage disorder arising from deficiency of iduronate-2-sulfatase (IDS), which results in progressive accumulation of glycosaminoglycans (GAGs) in multiple tissues. Accumulated GAGs are generally measured as the amount of total GAGs. However, we recently demonstrated that GAG accumulation in the brain of MPS II model mice cannot be reliably detected by conventional dye-binding assay measuring total GAGs. Here we developed a novel quantitative method for measurement of disease-specific GAGs based on the analysis of 2-sulfoiduronic acid levels derived from the non-reducing terminal end of the polysaccharides by using recombinant human IDS (rhIDS) and recombinant human iduronidase (rhIDUA). This method was evaluated on GAGs obtained from the liver and brain of MPS II mice. The GAGs were purified from tissue homogenates and then digested with rhIDS and rhIDUA to generate a desulfated iduronic acid from their non-reducing terminal end. HPLC analysis revealed that the generated iduronic acid levels were markedly increased in the liver and cerebrum of the MPS II mice, whereas the uronic acid was not detected in wild-type mice. These results indicate that this assay clearly detects the disease-specific GAGs in tissues from MPS II mice.
authors.affiliation: Division of Gene Therapy, Research Center for Medical Sciences, The Jikei University School of Medicine, Tokyo 105-8461, Japan. Electronic address: shimada_y@jikei.ac.jp.
authors.forename: Yohta
authors.initials: Y
authors.lastname: Shimada
authors.affiliation: Division of Gene Therapy, Research Center for Medical Sciences, The Jikei University School of Medicine, Tokyo 105-8461, Japan; Department of Pediatrics, The Jikei University School of Medicine, Tokyo 105-8461, Japan.
authors.forename: Taichi
authors.initials: T
authors.lastname: Wakabayashi
authors.affiliation: Division of Gene Therapy, Research Center for Medical Sciences, The Jikei University School of Medicine, Tokyo 105-8461, Japan; Department of Pediatrics, Kitasato University Graduate School of Medicine, Kanagawa, Japan.
authors.forename: Kazumasa
authors.initials: K
authors.lastname: Akiyama
authors.affiliation: Division of Gene Therapy, Research Center for Medical Sciences, The Jikei University School of Medicine, Tokyo 105-8461, Japan; Department of Pediatrics, The Jikei University School of Medicine, Tokyo 105-8461, Japan.
authors.forename: Hiroo
authors.initials: H
authors.lastname: Hoshina
authors.affiliation: Division of Gene Therapy, Research Center for Medical Sciences, The Jikei University School of Medicine, Tokyo 105-8461, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Higuchi
authors.affiliation: Division of Gene Therapy, Research Center for Medical Sciences, The Jikei University School of Medicine, Tokyo 105-8461, Japan; Department of Pediatrics, The Jikei University School of Medicine, Tokyo 105-8461, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kobayashi
authors.affiliation: Advanced Clinical Research Center, Institute of Neurological Disorders, Kanagawa, Japan.
authors.forename: Yoshikatsu
authors.initials: Y
authors.lastname: Eto
authors.affiliation: Division of Gene Therapy, Research Center for Medical Sciences, The Jikei University School of Medicine, Tokyo 105-8461, Japan; Department of Pediatrics, The Jikei University School of Medicine, Tokyo 105-8461, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Ida
authors.affiliation: Division of Gene Therapy, Research Center for Medical Sciences, The Jikei University School of Medicine, Tokyo 105-8461, Japan; Department of Pediatrics, The Jikei University School of Medicine, Tokyo 105-8461, Japan.
authors.forename: Toya
authors.initials: T
authors.lastname: Ohashi
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D054022:Cerebrum / Q000378:metabolism; D056947:Enzyme Replacement Therapy; D005260:Female; D006025:Glycosaminoglycans / Q000378:metabolism*; D006801:Humans; D007066:Iduronate Sulfatase / Q000737:chemistry / Q000627:therapeutic use; D007067:Iduronic Acid / Q000737:chemistry / Q000378:metabolism*; D007068:Iduronidase / Q000737:chemistry / Q000627:therapeutic use; D008099:Liver / Q000378:metabolism; D008810:Mice, Inbred C57BL; D016532:Mucopolysaccharidosis II / Q000175:diagnosis* / Q000188:drug therapy / Q000378:metabolism
chemical_list: D015415:Biomarkers; D006025:Glycosaminoglycans; D007067:Iduronic Acid; D007066:Iduronate Sulfatase; D007068:Iduronidase
keywords: Glycosaminoglycan; HPLC; Iduronic acid; MPS II; Mucopolysaccharidosis type II