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Biochem J. Jun 15, 2002; 364(Pt 3): 747–753.
PMCID: PMC1222624

Correction of mucopolysaccharidosis type IIIb fibroblasts by lentiviral vector-mediated gene transfer.


Mucopolysaccharidosis type IIIB (MPS IIIB; or Sanfilippo syndrome type B) is a lysosomal disease, due to glycosaminoglycan storage caused by mutations on the alpha-N-acetylglucosaminidase (NAGLU) gene. The disease is characterized by neurological dysfunction but relatively mild somatic manifestations. No effective treatment is available for affected patients. In the present study, we evaluated the role of a lentiviral vector as the transducing agent of NAGLU cDNA in MPS IIIB fibroblasts. The vector expressed high transduction efficiency and high levels of enzymic activity, 20-fold above normal levels, persisting for at least 2 months. PCR experiments confirmed the integration of the viral vector into the target genome. The NAGLU activity restored by virus infection was sufficient to normalize glycosaminoglycan accumulation, which is directly responsible for the disease phenotype. Metabolic labelling experiments on transduced fibroblasts exhibited, in the medium and in cellular lysates, polypeptide forms of 84 and 80 kDa respectively related to the precursor and mature forms of the enzyme. The enzyme secreted by transduced MPS IIIB fibroblasts was endocytosed in deficient cells by the mannose 6-phosphate system. Thus we show that lentiviral vectors may provide a therapeutic approach for the treatment of MPS IIIB disease.

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Selected References

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  • von Figura K. Human alpha-N-acetylglucosaminidase. 1. Purification and properties. Eur J Biochem. 1977 Nov 1;80(2):523–533. [PubMed]
  • Sasaki T, Sukegawa K, Masue M, Fukuda S, Tomatsu S, Orii T. Purification and partial characterization of alpha-N-acetylglucosaminidase from human liver. J Biochem. 1991 Nov;110(5):842–846. [PubMed]
  • Weber B, Blanch L, Clements PR, Scott HS, Hopwood JJ. Cloning and expression of the gene involved in Sanfilippo B syndrome (mucopolysaccharidosis III B). Hum Mol Genet. 1996 Jun;5(6):771–777. [PubMed]
  • von Figura K, Hasilik A, Steckel F, van de Kamp J. Biosynthesis and maturation of alpha-N-acetylglucosaminidase in normal and Sanfilippo B-fibroblasts. Am J Hum Genet. 1984 Jan;36(1):93–100. [PMC free article] [PubMed]
  • Di Natale P, Salvatore D, Daniele A, Bonatti S. Biosynthesis of alpha-N-acetylglucosaminidase in cultured human kidney carcinoma cells. Enzyme. 1985;33(2):75–83. [PubMed]
  • Zhao HG, Li HH, Bach G, Schmidtchen A, Neufeld EF. The molecular basis of Sanfilippo syndrome type B. Proc Natl Acad Sci U S A. 1996 Jun 11;93(12):6101–6105. [PMC free article] [PubMed]
  • Schmidtchen A, Greenberg D, Zhao HG, Li HH, Huang Y, Tieu P, Zhao HZ, Cheng S, Zhao Z, Whitley CB, et al. NAGLU mutations underlying Sanfilippo syndrome type B. Am J Hum Genet. 1998 Jan;62(1):64–69. [PMC free article] [PubMed]
  • Zhao HG, Aronovich EL, Whitley CB. Genotype-phenotype correspondence in Sanfilippo syndrome type B. Am J Hum Genet. 1998 Jan;62(1):53–63. [PMC free article] [PubMed]
  • Beesley CE, Young EP, Vellodi A, Winchester BG. Identification of 12 novel mutations in the alpha-N-acetylglucosaminidase gene in 14 patients with Sanfilippo syndrome type B (mucopolysaccharidosis type IIIB). J Med Genet. 1998 Nov;35(11):910–914. [PMC free article] [PubMed]
  • Bunge S, Knigge A, Steglich C, Kleijer WJ, van Diggelen OP, Beck M, Gal A. Mucopolysaccharidosis type IIIB (Sanfilippo B): identification of 18 novel alpha-N-acetylglucosaminidase gene mutations. J Med Genet. 1999 Jan;36(1):28–31. [PMC free article] [PubMed]
  • Yogalingam G, Hopwood JJ. Molecular genetics of mucopolysaccharidosis type IIIA and IIIB: Diagnostic, clinical, and biological implications. Hum Mutat. 2001 Oct;18(4):264–281. [PubMed]
  • Tessitore A, Villani GR, Di Domenico C, Filocamo M, Gatti R, Di Natale P. Molecular defects in the alpha-N-acetylglucosaminidase gene in Italian Sanfilippo type B patients. Hum Genet. 2000 Dec;107(6):568–576. [PubMed]
  • Zhao KW, Neufeld EF. Purification and characterization of recombinant human alpha-N-acetylglucosaminidase secreted by Chinese hamster ovary cells. Protein Expr Purif. 2000 Jun;19(1):202–211. [PubMed]
  • Yogalingam G, Weber B, Meehan J, Rogers J, Hopwood JJ. Mucopolysaccharidosis type IIIB: characterisation and expression of wild-type and mutant recombinant alpha-N-acetylglucosaminidase and relationship with sanfilippo phenotype in an attenuated patient. Biochim Biophys Acta. 2000 Nov 15;1502(3):415–425. [PubMed]
  • Weber B, Hopwood JJ, Yogalingam G. Expression and characterization of human recombinant and alpha-N-acetylglucosaminidase. Protein Expr Purif. 2001 Mar;21(2):251–259. [PubMed]
  • Li HH, Yu WH, Rozengurt N, Zhao HZ, Lyons KM, Anagnostaras S, Fanselow MS, Suzuki K, Vanier MT, Neufeld EF. Mouse model of Sanfilippo syndrome type B produced by targeted disruption of the gene encoding alpha-N-acetylglucosaminidase. Proc Natl Acad Sci U S A. 1999 Dec 7;96(25):14505–14510. [PMC free article] [PubMed]
  • Naldini L, Verma IM. Lentiviral vectors. Adv Virus Res. 2000;55:599–609. [PubMed]
  • Buchschacher GL, Jr, Wong-Staal F. Development of lentiviral vectors for gene therapy for human diseases. Blood. 2000 Apr 15;95(8):2499–2504. [PubMed]
  • Nègre D, Mangeot PE, Duisit G, Blanchard S, Vidalain PO, Leissner P, Winter AJ, Rabourdin-Combe C, Mehtali M, Moullier P, et al. Characterization of novel safe lentiviral vectors derived from simian immunodeficiency virus (SIVmac251) that efficiently transduce mature human dendritic cells. Gene Ther. 2000 Oct;7(19):1613–1623. [PubMed]
  • Johnson LG, Olsen JC, Naldini L, Boucher RC. Pseudotyped human lentiviral vector-mediated gene transfer to airway epithelia in vivo. Gene Ther. 2000 Apr;7(7):568–574. [PubMed]
  • Wang G, Sinn PL, McCray PB., Jr Development of retroviral vectors for gene transfer to airway epithelia. Curr Opin Mol Ther. 2000 Oct;2(5):497–506. [PubMed]
  • Richter J, Karlsson S. Clinical gene therapy in hematology: past and future. Int J Hematol. 2001 Feb;73(2):162–169. [PubMed]
  • Pfeifer A, Kessler T, Yang M, Baranov E, Kootstra N, Cheresh DA, Hoffman RM, Verma IM. Transduction of liver cells by lentiviral vectors: analysis in living animals by fluorescence imaging. Mol Ther. 2001 Mar;3(3):319–322. [PubMed]
  • Bosch A, Perret E, Desmaris N, Trono D, Heard JM. Reversal of pathology in the entire brain of mucopolysaccharidosis type VII mice after lentivirus-mediated gene transfer. Hum Gene Ther. 2000 May 20;11(8):1139–1150. [PubMed]
  • Consiglio A, Quattrini A, Martino S, Bensadoun JC, Dolcetta D, Trojani A, Benaglia G, Marchesini S, Cestari V, Oliverio A, et al. In vivo gene therapy of metachromatic leukodystrophy by lentiviral vectors: correction of neuropathology and protection against learning impairments in affected mice. Nat Med. 2001 Mar;7(3):310–316. [PubMed]
  • Follenzi A, Ailles LE, Bakovic S, Geuna M, Naldini L. Gene transfer by lentiviral vectors is limited by nuclear translocation and rescued by HIV-1 pol sequences. Nat Genet. 2000 Jun;25(2):217–222. [PubMed]
  • Dull T, Zufferey R, Kelly M, Mandel RJ, Nguyen M, Trono D, Naldini L. A third-generation lentivirus vector with a conditional packaging system. J Virol. 1998 Nov;72(11):8463–8471. [PMC free article] [PubMed]
  • Sonza S, Maerz A, Deacon N, Meanger J, Mills J, Crowe S. Human immunodeficiency virus type 1 replication is blocked prior to reverse transcription and integration in freshly isolated peripheral blood monocytes. J Virol. 1996 Jun;70(6):3863–3869. [PMC free article] [PubMed]
  • Follenzi A, Sabatino G, Lombardo A, Boccaccio C, Naldini L. Efficient gene delivery and targeted expression to hepatocytes in vivo by improved lentiviral vectors. Hum Gene Ther. 2002 Jan 20;13(2):243–260. [PubMed]
  • Chow P, Weissmann B. 4-Methylumbelliferyl 2-acetamido-2-deoxy-alpha-D-glucopyranoside, a fluorogenic substrate for N-acetyl-alpha-D-glucosaminidase. Carbohydr Res. 1981 Oct 1;96(1):87–93. [PubMed]
  • Dahms NM, Lobel P, Kornfeld S. Mannose 6-phosphate receptors and lysosomal enzyme targeting. J Biol Chem. 1989 Jul 25;264(21):12115–12118. [PubMed]
  • Vellodi A, Young E, New M, Pot-Mees C, Hugh-Jones K. Bone marrow transplantation for Sanfilippo disease type B. J Inherit Metab Dis. 1992;15(6):911–918. [PubMed]
  • Bielicki J, Hopwood JJ, Anson DS. Correction of Sanfilippo A skin fibroblasts by retroviral vector-mediated gene transfer. Hum Gene Ther. 1996 Oct 20;7(16):1965–1970. [PubMed]
  • Braun SE, Aronovich EL, Anderson RA, Crotty PL, McIvor RS, Whitley CB. Metabolic correction and cross-correction of mucopolysaccharidosis type II (Hunter syndrome) by retroviral-mediated gene transfer and expression of human iduronate-2-sulfatase. Proc Natl Acad Sci U S A. 1993 Dec 15;90(24):11830–11834. [PMC free article] [PubMed]
  • Palmer TD, Rosman GJ, Osborne WR, Miller AD. Genetically modified skin fibroblasts persist long after transplantation but gradually inactivate introduced genes. Proc Natl Acad Sci U S A. 1991 Feb 15;88(4):1330–1334. [PMC free article] [PubMed]
  • Challita PM, Skelton D, el-Khoueiry A, Yu XJ, Weinberg K, Kohn DB. Multiple modifications in cis elements of the long terminal repeat of retroviral vectors lead to increased expression and decreased DNA methylation in embryonic carcinoma cells. J Virol. 1995 Feb;69(2):748–755. [PMC free article] [PubMed]
  • Wolfe JH, Sands MS, Barker JE, Gwynn B, Rowe LB, Vogler CA, Birkenmeier EH. Reversal of pathology in murine mucopolysaccharidosis type VII by somatic cell gene transfer. Nature. 1992 Dec 24;360(6406):749–753. [PubMed]
  • Daly TM, Vogler C, Levy B, Haskins ME, Sands MS. Neonatal gene transfer leads to widespread correction of pathology in a murine model of lysosomal storage disease. Proc Natl Acad Sci U S A. 1999 Mar 2;96(5):2296–2300. [PMC free article] [PubMed]
  • Dittmer F, Ulbrich EJ, Hafner A, Schmahl W, Meister T, Pohlmann R, von Figura K. Alternative mechanisms for trafficking of lysosomal enzymes in mannose 6-phosphate receptor-deficient mice are cell type-specific. J Cell Sci. 1999 May;112(Pt 10):1591–1597. [PubMed]
  • Yu WH, Zhao KW, Ryazantsev S, Rozengurt N, Neufeld EF. Short-term enzyme replacement in the murine model of Sanfilippo syndrome type B. Mol Genet Metab. 2000 Dec;71(4):573–580. [PubMed]
  • Hoogerbrugge PM, Suzuki K, Suzuki K, Poorthuis BJ, Kobayashi T, Wagemaker G, van Bekkum DW. Donor-derived cells in the central nervous system of twitcher mice after bone marrow transplantation. Science. 1988 Feb 26;239(4843):1035–1038. [PubMed]
  • Shi N, Pardridge WM. Noninvasive gene targeting to the brain. Proc Natl Acad Sci U S A. 2000 Jun 20;97(13):7567–7572. [PMC free article] [PubMed]

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