Metabolic Correction and Cross-Correction of Mucopolysaccharidosis Type II (Hunter Syndrome) by Retroviral-Mediated Gene Transfer and Expression of Human Iduronate-2-Sulfatase

• Published in: Proceedings of the National Academy of Sciences - PNAS 1993-12, Vol.90 (24), p.11830-11834
• Main Authors: Braun, Stephen E., Aronovich, Elena L., Anderson, Rose Ann, Crotty, Paul L., McIvor, R. Scott, Whitley, Chester B.
• Format: Article
• Language: English
• Subjects: Base Sequence; Biological and medical sciences; Biotechnology; Blotting, Southern; Cell Line; Cell lines; Cells; Cells, Cultured; Complementary DNA; Cultured cells; DNA; DNA - genetics; DNA - isolation & purification; DNA Primers; Enzyme activity; Enzyme substrates; Enzymes; Fibroblasts; Fibroblasts - enzymology; Fundamental and applied biological sciences. Psychology; Gene Expression; Gene therapy; Gene Transfer Techniques; Genetic Therapy - methods; Genetic Vectors; Genetics; Glycosaminoglycans - biosynthesis; Health. Pharmaceutical industry; Humans; Iduronate Sulfatase - genetics; Iduronate Sulfatase - metabolism; Industrial applications and implications. Economical aspects; Lymphocytes; Medical disorders; Medical research; Metabolism; Molecular Sequence Data; Mucopolysaccharidosis II - enzymology; Mucopolysaccharidosis II - genetics; Mucopolysaccharidosis II - therapy; Polymerase Chain Reaction - methods; Restriction Mapping; Retroviridae - genetics; Sulfates - metabolism
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.90.24.11830

To explore the possibility of using gene transfer to provide iduronate-2-sulfatase (IDS; EC 3.1.6.13) enzyme activity for treatment of Hunter syndrome, an amphotropic retroviral vector, L2SN, containing the human IDS coding sequence was constructed and studied for gene expression in vitro. Lymphoblastoid cell lines (LCLs) from patients with Hunter syndrome were transduced with L2SN and expressed high levels of IDS enzyme activity, 10- to 70-fold higher than normal human peripheral blood leukocytes or LCLs. Such L2SN-transduced LCLs failed to show accumulation of35SO4into glycosaminoglycan (35SO4-GAG), indicating that recombinant IDS enzyme participated in GAG metabolism. Coculture of L2SN-transduced LCLs with fibroblasts from patients with Hunter syndrome reduced the accumulation of35SO4-GAG. These results demonstrated retroviral-mediated IDS gene transfer into lymphoid cells and the ability of such cells to provide recombinant enzyme for intercellular metabolic cross-correction.