Mucopolysaccharidosis type IIIB (MPS IIIB; or Sanfilippo syndrome type B) is a lysosomal disease, due to glycosaminoglycan storage caused by mutations on the α-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 80kDa 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.

Abbreviations used: BMT, bone marrow transplantation; CMV, the immediate early enhancer/promoter of the human cytomegalovirus; CNS, central nervous system; DMEM, Dulbecco's modified Eagle's medium; FCS, fetal calf serum; GAG, glycosaminoglycan; GFP, green fluorescent protein, MPS, mucopolysaccharidosis; NAGLU, α-N-acetylglucosaminidase; PPT, polypurine tract and termination sequences from pol of HIV-1-enhancing nuclear translocation; WPRE, the post-transcriptional regulatory element from the woodchuck hepatitis virus.

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