METHODS AND COMPOSITIONS FOR THE TREATMENT OF LYSOSOMAL STORAGE DISEASES

• US 20160060656A1
• Filed: 10/01/2015
• Published: 03/03/2016
• Est. Priority Date: 07/11/2012
• Status: Active Grant

• Current Assignee: Sangamo Therapeutics, Inc.
• Original Assignee: Sangamo Biosciences, Inc.
• Inventors: Rebar, Edward J.
• Granted Patent: US 10,648,001 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61K 35/407 Liver; Hepatocytes
• CPC Class Codes: A61K 38/465 acting on ester bonds (3.1), e.g. lipases, ribonucleases
• CPC Class Codes: A61K 38/47 acting on glycosyl compounds (3.2), e.g. cellulases, lactases
• 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: C07K 2319/81 containing a Zn-finger domain for DNA binding
• CPC Class Codes: C12N 15/907 in mammalian cells
• CPC Class Codes: C12N 9/22 Ribonucleases RNAses, DNAses catalytic nucleic acids C12N15/113
• CPC Class Codes: C12Y 301/06013 Iduronate-2-sulfatase (3.1.6.13)
• CPC Class Codes: C12Y 302/0102 Alpha-glucosidase (3.2.1.20)
• CPC Class Codes: C12Y 302/01021 Beta-glucosidase (3.2.1.21)
• CPC Class Codes: C12Y 302/01022 Alpha-galactosidase (3.2.1.22)
• CPC Class Codes: C12Y 302/01076 L-Iduronidase (3.2.1.76)

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 treatment and/or prevention of a lysosomal storage disease.

Claims

1. A method for generating a liver cell that produces a protein, the method comprising:

providing to the liver cell at least one expression vector comprising a transgene encoding the protein and a sequence encoding a non-naturally occurring nuclease that cleaves an endogenous HPRT locus in the liver cell, such that the transgene is integrated into the endogenous HPRT locus of the liver cell and the liver cell produces the protein, wherein the protein is α

-galactosidase A (GLA), acid β

-glucosidase (GBA), α

-L-iduronidase (IDUA), alpha-glucosidase (GAA) or iduronate-2-sulfatase (IDS).

2. The method of claim 1, wherein the protein is lacking or deficient in a disease selected from the group consisting of Gaucher'"'"'s, Fabry'"'"'s, Hunter'"'"'s, Pompe'"'"'s and Hurler'"'"'s.

3. The method of claim 1, wherein expression of the transgene is driven by an endogenous 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 HPRT locus into which the transgene is integrated.

5. The method of claim 1, wherein the transgene encodes a fusion protein comprising an extracellular domain of a membrane protein such that upon expression, the fusion protein is localized on the surface of the cell.

6. The method of claim 1, wherein the transgene encodes a fusion protein comprising a ligand for a receptor.

7. A method of treating a lysosomal storage disease, the method comprising

isolating the protein produced by the cell generated according to the method of claim 1, and

administering the protein to a patient in need thereof.

8. A method of treating a lysosomal storage disease, the method comprising

administering the cell generated by the method of claim 1 to a subject in need thereof.

9. A method of treating a lysosomal storage disease, the method comprising

administering one or more nucleases and one or more transgenes to a subject in need thereof, such that a cell generated according to the method of claim 1 is generated in the subject.

10. The method of claim 9, wherein the cell is an isolated stem or precursor cell, and the method further comprises expanding and differentiating the stem or precursor cell prior to administration.

11. The method of claim 1, wherein the transgene is delivered to the cell using a viral vector.

12. The method of claim 11, wherein the viral vector is an AAV vector.

13. A cell produced by the method of claim 1, wherein the transgene encoding the protein is integrated into the endogenous HPRT gene within 1 to 300 base pairs of SEQ ID NO:

24 and/or 25.