Diagnosis and treatment of multiple sulfatase deficiency and other sulfatase deficiencies

Abstract

This invention relates to methods and compositions for the diagnosis and treatment of Multiple Sulfatase Deficiency (MSD) as well as other sulfatase deficiencies. More specifically, the invention relates to isolated molecules that modulate post-translational modifications on sulfatases. Such modifications are essential for proper sulfatase function.

1 .- 85 . (canceled) 
     
     
         86 . A composition comprising isolated recombinant Iduronate 2-Sulfatase produced by cultured mammalian cells transfected with a vector comprising a heterologous polynucleotide encoding Iduronate 2-Sulfatase (I2S) and wherein the mammalian cells express a Formylglycine Generating Enzyme (FGE) such that the ratio of active I2S to total I2S in the composition is increased by at least 5% compared to the ratio of active I2S to total I2S produced by cells without expression of FGE; wherein the active I2S comprises a Formylglycine; and
 wherein the FGE comprises an amino acid sequence having at least 95% identity to the amino acid sequence of amino acids 34-374 of SEQ ID NO:2 and has Cα-formylglycine generating activity.   
     
     
         87 . The composition of  claim 86 , wherein the FGE is endogenous to the cultured mammalian cells. 
     
     
         88 . The composition of  claim 86 , wherein the FGE is exogenous to the cultured mammalian cells. 
     
     
         89 . The composition of  claim 86 , wherein the mammalian cells are further transfected with an expression vector comprising a polynucleotide encoding the formylglycine generating enzyme (FGE) and wherein the mammalian cells express the FGE as compared to mammalian cells that are not transfected with the vector comprising a polynucleotide encoding the FGE. 
     
     
         90 . The composition of  claim 86 , wherein the recombinant Iduronate 2-Sulfatase is encoded by a nucleic acid comprising the nucleotide sequence of SEQ ID NO: 6. 
     
     
         91 . The composition of  claim 86 , wherein the recombinant Iduronate 2-Sulfatase comprises the amino acid sequence of SEQ ID NO: 7. 
     
     
         92 . The composition of  claim 86 , wherein the recombinant Iduronate-2-Sulfatase has a specific activity of 3-5 U/mg as measured by an ELISA-based immunoassay. 
     
     
         93 . A method of treating Hunter Syndrome in a subject, comprising administering into the subject a pharmaceutical composition comprising the composition of  claim 86 . 
     
     
         94 . A composition comprising isolated recombinant Iduronate 2-Sulfatase produced by cultured mammalian cells transfected with a vector comprising a heterologous polynucleotide encoding Iduronate 2-Sulfatase (I2S) and wherein the mammalian cells express a Formylglycine Generating Enzyme (FGE) such that the ratio of active I2S to total I2S in the composition is increased by at least 5% compared to the ratio of active I2S to total I2S produced by cells without expression of FGE; wherein the active I2S comprises a Formylglycine; and
 wherein the FGE comprises an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO:50 and has Cα-formylglycine generating activity.   
     
     
         95 . The composition of  claim 94 , wherein the FGE is endogenous to the cultured mammalian cells. 
     
     
         96 . The composition of  claim 94 , wherein the FGE is exogenous to the cultured mammalian cells. 
     
     
         97 . The composition of  claim 94 , wherein the recombinant Iduronate 2-Sulfatase is encoded by a nucleic acid comprising the nucleotide sequence of SEQ ID NO: 6. 
     
     
         98 . The composition of  claim 94 , wherein the recombinant Iduronate 2-Sulfatase comprises the amino acid sequence of SEQ ID NO: 7. 
     
     
         99 . A gene therapy method of treating Sanfilippo Syndrome A, the method comprising delivering to a subject in need of treatment
 a vector comprising a polynucleotide encoding a Formylglycine Generating Enzyme (FGE) comprising an amino acid sequence having at least 80% identity to the amino acid sequence of amino acids 34-374 of SEQ ID NO:2, wherein the FGE has C α -formylglycine generating activity; and   a vector comprising a polynucleotide encoding Sulfamidase such that one or more symptoms of Sanfilippo Syndrome A is ameliorated, reduced or eliminated.   
     
     
         100 . The gene therapy method of  claim 99 , wherein the polynucleotide encoding a Formylglycine Generating Enzyme (FGE) and the polynucleotide encoding Sulfamidase are delivered on separate vectors. 
     
     
         101 . The gene therapy method of  claim 99 , wherein the polynucleotide encoding a Formylglycine Generating Enzyme (FGE) and the polynucleotide encoding Sulfamidase are delivered on the same vector. 
     
     
         102 . The gene therapy method of  claim 99 , wherein the vector comprising a polynucleotide encoding a Formylglycine Generating Enzyme (FGE) and/or the vector comprising a polynucleotide encoding Sulfamidase is a viral vector. 
     
     
         103 . The gene therapy method of  claim 102 , wherein the delivering step comprises in vivo transduction of the cell of the subject. 
     
     
         104 . The gene therapy method of  claim 103 , wherein the cell is selected from the group consisting of fibroblasts, keratinocytes, epithelial cells, endothelial cells, glial cells, neural cells, blood cells, lymphocytes, bone marrow cells, muscle cells, and precursors thereof. 
     
     
         105 . The gene therapy method of  claim 99 , wherein the polynucleotide encoding the FGE and/or the polynucleotide encoding Sulfamidase is operably linked to a gene expression regulatory sequence.