title: Prediction of Sanfilippo phenotype severity from immunoquantification of heparan-N-sulfamidase in cultured fibroblasts from mucopolysaccharidosis type IIIA patients.
abstract: Mucopolysaccharidosis type IIIA (MPS-IIIA) is an autosomal recessive lysosomal storage disorder caused by the deficiency of heparan-N-sulfamidase (NS; EC 3.10.1.1), resulting in defective degradation and subsequent storage of heparan sulfate and leading to a clinical phenotype known as Sanfilippo syndrome. A sensitive and specific monoclonal/polyclonal-based immunoquantification assay has enabled the determination of NS protein, down to approximately 3 pg NS protein, in cultured fibroblasts from control and MPS-IIIA patients. Cultured skin fibroblasts from 15 normal controls contained 11.9 to 105 ng of NS protein/mg extracted cell protein, whereas NS protein ranged from "none detected" to 11 ng/mg in fibroblasts from 35 MPS-IIIA patients. A relationship between genotype/phenotype and amount of NS protein present in these MPS-IIIA fibroblasts was established. Immunoquantification, in combination with a specific and highly sensitive tetrasaccharide-based assay of NS activity, enabled the determination of residual specific NS activity in these fibroblasts. Specific NS activity ranged from 28 to 1289 nmol/min/mg NS protein for MPS-IIIA patients, compared to 870 nmol/min/mg of recombinant human NS. It is proposed that this immunoquantification method, in conjunction with the specific NS activity assay, may be used to predict clinical severity in MPS-IIIA patients, allowing for the selection of individuals best suited for gene- and enzyme-replacement therapy when these methods become available. Also proposed is that an enzyme-replacement therapy achieving a correction of approximately 10% of normal NS activity is required to avoid the onset of a Sanfilippo clinical phenotype.
authors.affiliation: Lysosomal Storage Research Unit, Department of Chemical Pathology, Women's and Children's Hospital, 72 King William Road, North Adelaide, South Australia, 5006, Australia.
authors.forename: K J
authors.initials: KJ
authors.lastname: Perkins
authors.forename: V
authors.initials: V
authors.lastname: Muller
authors.forename: B
authors.initials: B
authors.lastname: Weber
authors.forename: J J
authors.initials: JJ
authors.lastname: Hopwood
mesh_terms: D000293:Adolescent; D000328:Adult; D002478:Cells, Cultured; D002675:Child, Preschool; D004795:Enzyme Stability; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005347:Fibroblasts / Q000166:cytology / Q000201:enzymology; D006801:Humans; D006867:Hydrolases / Q000032:analysis / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D007223:Infant; D009084:Mucopolysaccharidosis III / Q000175:diagnosis / Q000201:enzymology* / Q000473:pathology / Q000503:physiopathology*; D009154:Mutation; D010641:Phenotype; D012015:Reference Standards
chemical_list: D006867:Hydrolases; C022708:N-sulfoglucosamine sulfohydrolase