Method of treating lysosomal storage diseases using nucleases and a transgene
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 for expressing and secreting exogenous glucocerbrosidase (GBA), α-galactosidase A (GLA), iduronate sulftase (IDS), or iduronidase (IDUA) protein in a liver cell in a mouse or human with a deficient GLA, GBA, IDS, or IDUA gene, the method comprising:
(i) intravenously injecting one or more adeno-associated viral (AAV) vectors encoding a pair of nucleases into the mouse or human, wherein the nucleases cleave an endogenous albumin gene in the liver cell; and
(ii) intravenously injecting an AAV vector comprising a donor sequence comprising a transgene encoding:
(a) an exogenous GBA protein into the mouse or human with the deficient GBA gene;
(b) an exogenous GLA protein into the mouse or human with the deficient GLA gene;
(c) an exogenous IDS protein into the mouse or human with the deficient IDS gene; or
(d) an exogenous IDUA protein into the mouse or human with the deficient IDUA gene, and
wherein the transgene is flanked by sequences having homology with the endogenous albumin gene,
such that the transgene is integrated into the endogenous albumin gene in the liver cell, and the liver cell expresses and secretes the exogenous GBA, GLA, IDS, or IDUA protein.
2. The method of claim 1 , wherein expression of the transgene is driven by the endogenous albumin promoter.
3. A method of treating a mouse or human with Gaucher's, Fabry's, Hunter's or Hurler's disease, the method comprising:
expressing and secreting exogenous GBA protein in a liver cell in the mouse or human with Gaucher's disease,
expressing and secreting exogenous GLA protein in a liver cell in the mouse or human with Fabry's disease,
expressing and secreting exogenous IDS protein in a liver cell in the mouse or human with Hunter's/MPS II disease, or
expressing and secreting exogenous IDUA protein in a liver cell in the mouse or human with Hurler's/MPS I disease
according to the method of claim 1 such that therapeutic levels of the protein are obtained in the serum of the mouse or human.