METHODS AND COMPOSITIONS FOR THE PROVISION OF PROTEINS DEFICIENT IN LYSOSOMAL STORAGE DISEASES

• US 20140112896A1
• Filed: 07/11/2013
• Published: 04/24/2014
• Est. Priority Date: 07/11/2012
• Status: Active Grant

• Current Assignee: Sangamo Therapeutics, Inc.
• Original Assignee: Sangamo Biosciences Incorporated
• Inventors: Rebar, Edward J.
• Granted Patent: US 9,956,247 B2
• Time in Patent Office: Days
• US Class Current: 424/93.21
• 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 generating a cell that produces a protein deficient in a lysosomal storage disease, the method comprising:

integrating a transgene encoding the protein into an endogenous locus of the cell using a non-naturally occurring nuclease, such that the cell produces the protein that is deficient in the lysosomal storage disease.

2. The method of claim 1, wherein the lysosomal storage disease is selected from the group consisting of Gaucher'"'"'s, Fabry'"'"'s, Hunter'"'"'s, Hurler'"'"'s, and Niemann-Pick'"'"'s.

3. The method of claim 1, wherein the transgene encodes a protein selected from the group consisting of glucocerebrosidase, α

galactosidase, iduronate-2-sulfatase deficiency, alpha-L iduronidase deficiency and sphingomyelin phosphodiesterase.

4. The method of claim 1, wherein the endogenous locus is selected from the group consisting of an albumin gene, an AAVS1 gene, an HRPT gene, a CCR5 gene, a Rosa gene and a globin gene.

5. The method of claim 1, wherein expression of the transgene is driven by an endogenous promoter.

6. The method of claim 1, wherein the transgene is a fusion protein comprising amino acids encoded by the transgene and by the endogenous locus into which the transgene is integrated.

7. 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.

8. The method of claim 1, wherein the transgene encodes a fusion protein comprising a ligand for a receptor such that upon expression, the fusion protein crosses the blood brain barrier.

9. The method of claim 1, wherein the cell is selected from the group consisting of a red blood cell, a liver cell, a muscle cell and a stem cell.

10. The method of claim 9, wherein the stem cell is a hematopoietic stem cell or induced pluripotent stem cell.

11. A method of providing a protein deficient in 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.

12. A method of providing a protein deficient in a lysosomal storage disease, the method comprising

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

13. A method of providing a protein deficient in 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.

14. The method of claim 12, 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.

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

16. The method of claim 15, wherein the viral vector is an AAV vector.