Methods of treating lysosomal storage diseases

Abstract

Nucleases and methods of using these nucleases for inserting a sequence encoding a therapeutic protein such as an enzyme into a cell, thereby providing proteins or cell therapeutics for the provision of proteins lacking or deficient in subjects with a lysosomal storage disease and treatment and/or prevention of lysosomal storage diseases.

What is claimed is: 
     
         1 . A method for expressing and secreting a glucocerbrosidase (GBA) protein, an α-galactosidase A (GLA) protein, an iduronate sulfatase protein (IDS), an iduronidase (IDUA) protein, an α-Glucosidase (GAA) protein or a sphingomyelin protein in a liver cell of a subject in need thereof, the method comprising:
 (a) administering one or more polynucleotides comprising an artificial nuclease that cleaves intron 12 of an endogenous albumin gene into said subject containing the liver cell; and 
 (b) administering a donor sequence comprising a transgene encoding a GBA, GLA, IDS, IDUA, GAA or spingomyelin protein into the liver cell of the subject, such liver cell expresses and secretes the exogenous GBA, GLA, IDS, or IDUA protein. 
 
     
     
         2 . The method of  claim 1 , wherein the transgene encodes iduronate-2-sulfatase or alpha-L iduronidase. 
     
     
         3 . The method of  claim 1 , wherein expression of the transgene is driven by the endogenous albumin promoter. 
     
     
         4 . The method of  claim 1 , wherein the transgene encodes a fusion protein comprising amino acids encoded by the transgene and by the endogenous locus into which the transgene is integrated. 
     
     
         5 . The method of  claim 1 , wherein the artificial nuclease and donor sequence are co-administered. 
     
     
         6 . The method of  claim 1 , wherein expression of the transgene is driven by the endogenous albumin promoter. 
     
     
         7 . The method of  claim 1 , wherein the protein is secreted from the liver into the blood, spleen, kidney, lymph nodes and/or brain of the human subject. 
     
     
         8 . The method of  claim 1 , wherein the artificial nuclease comprises a zinc finger nuclease (ZFN), a TAL-effector domain nuclease (TALEN) or a CRISPR/Cas nuclease. 
     
     
         9 . A method of treating a human subject with a lysosomal storage disease in a human subject, the method comprising
 providing a protein to the human subject according to the method of  claim 1 , such that the lysosomal storage disease is treated.