Methods and compositions for the treatment of lysosomal storage diseases
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 treatment and/or prevention of a lysosomal storage disease.
1. A method of treating a mouse or human with Gaucher's, Fabry's, or Pompe's disease, the method comprising
intravenously injecting first and second AAV vectors encoding first and second ZFNs of a pair of zinc finger nucleases that cleave in an endogenous albumin gene, into the mouse or human with Gaucher's, Fabry's, or Pompe's disease;
intravenously injecting a third AAV vector comprising a transgene encoding:
(a) a glucocerebrosidase (GBA) protein into the mouse or human with Gaucher's disease;
(b) an α galactosidase deficiency (GLA) protein into the mouse or human with Fabry's disease;
(c) an alpha-glucosidase (GAA) protein into the mouse or human with Pompe's disease; or
wherein the transgene is flanked by sequences having homology with the endogenous albumin gene;
wherein the first, second and third AAV vectors are delivered at a ratio of 1:1:8 and further wherein the transgene is integrated into the endogenous albumin gene in liver cells of the mouse or human and the liver cells express and secrete therapeutic amounts of the protein and a symptom of the Gaucher's, Fabry's, or Pompe's disease is treated in the mouse or human.
2. The method of claim 1 , wherein the method treats neurological deficits associated with Gaucher's, Fabry's, or Pompe's disease.
3. The method of claim 1 , wherein expression of the transgene is driven by an endogenous albumin promoter.
4. The method of claim 1 , wherein the AAV vectors are AAV2 and/or AAV6 vectors.
5. The method of claim 1 , wherein the protein is detectable in secondary tissues in the mouse or human subject.
6. The method of claim 5 , wherein the secondary tissue is spleen.
7. The method of claim 5 , wherein the secondary tissue is blood or plasma.
8. The method of claim 5 , wherein the secondary tissue is kidney or lung.