Method of treating lysosomal storage diseases

• US 9,956,247 B2
• Filed: 07/11/2013
• Issued: 05/01/2018
• Est. Priority Date: 07/11/2012
• Status: Active Grant

• Current Assignee: Sangamo Therapeutics, Inc.
• Original Assignee: Sangamo Therapeutics, Inc.
• Inventors: Rebar, Edward J.
• Primary Examiner(s): SINGH, ANOOP KUMAR
• Application Number:
  US13/939,794
• Publication Number:
  US 20140112896A1
• Time in Patent Office: 1,755 Days
• CPC Class Codes: A61K 35/28 Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
• CPC Class Codes: A61K 38/46 Hydrolases (3)
• CPC Class Codes: A61K 38/465 acting on ester bonds (3.1), e.g. lipases, ribonucleases
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61K 48/005 characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 2319/81 containing a Zn-finger domain for DNA binding
• CPC Class Codes: C12N 15/8645 Adeno-associated virus
• CPC Class Codes: C12N 15/907 in mammalian cells
• CPC Class Codes: C12N 9/22 Ribonucleases RNAses, DNAses catalytic nucleic acids C12N15/113

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.

Claims

1. A method for expressing and secreting exogenous glucocerebrosidase (GBA), α

galactosidase (GLA), iduronate-2-sulfatase (IDS) or alpha-L iduronidase (IUDA) protein in a liver cell of a human or a mouse in need thereof, the method comprising;

(a) intravenously administering one or more AAV vectors encoding a pair of zinc finger nucleases that cleaves in intron 1 or 12 of an endogenous albumin gene into said human or mouse containing the liver cell and disrupts expression of the albumin gene; and

(b) intravenously administering an AAV vector comprising a donor sequence comprising a transgene encoding;

(i) an exogenous GBA protein;

(ii) an exogenous GLA protein;

(iii) an exogenous IDS protein;

or (iv) an exogenous IDUA protein, into the human or mouse liver cell of said human or mouse, wherein the transgene encoding the protein is flanked by regions of homology to intron 1 or 12 of the cleaved endogenous albumin gene, such that the transgene is integrated by homology directed repair into intron 1 or 12 of the cleaved endogenous albumin gene, and the 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.