METHODS AND COMPOSITIONS FOR DELIVERY OF BIOLOGICS

• US 20140017214A1
• Filed: 07/11/2013
• Published: 01/16/2014
• Est. Priority Date: 07/11/2012
• Status: Active Grant

• Current Assignee: Sangamo Therapeutics, Inc.
• Original Assignee: Sangamo Biosciences, Inc. (Sangamo Therapeutics, Inc.)
• Inventors: Cost, Gregory J.
• Granted Patent: US 10,883,119 B2
• Time in Patent Office: Days
• US Class Current: 424/93.21
• CPC Class Codes: A61K 35/18 Erythrocytes haemoglobin A61K38/42
• CPC Class Codes: A61K 38/465 acting on ester bonds (3.1), e.g. lipases, ribonucleases
• CPC Class Codes: A61P 39/02 Antidotes
• CPC Class Codes: C12N 15/907 in mammalian cells
• CPC Class Codes: C12Y 301/01008 Cholinesterase (3.1.1.8), i.e. butyrylcholine-esterase

Abstract

Nucleases and methods of using these nucleases for genetic alteration of red blood cells (RBCs), for example for providing for a protein lacking in a monogenic disorder or a biologic for the treatment of exposure to a toxin using genetically altered RBCs.

Claims

1. A genetically modified red blood cell (RBC) or RBC precursor cell comprising an integrated transgene that expresses a protein, wherein the protein degrades or detoxifies one or more toxins or supplies a protein lacking in a lysosomal storage disease.

2. The RBC or RBC precursor cell of claim 1, wherein the transgene is integrated into a globin or a safe harbor gene.

3. The RBC or RBC precursor cell of claim 2, wherein the globin gene is a beta globin gene or a gamma globin gene.

4. The RBC or RBC precursor cell of claim 1, wherein the protein is retained within the cell.

5. The RBC or RBC precursor cell of claim 1, wherein the transgene comprises an extracellular domain or a membrane protein and the protein is localized on the surface of the cell.

6. The RBC or RBC precursor cell of claim 1, wherein the transgene is expressed with endogenous sequences.

7. The RBC or RBC precursor cell of claim 6, wherein the endogenous sequences are present on the amino (N)-terminal portion or carboxy (C)-terminal portion of the transgene protein.

8. The RBC or RBC precursor cell of claim 1, wherein the transgene is integrated into the genome of the RBC or the RBC precursor cell following cleavage of the genome by a nuclease.

9. The RBC or RBC precursor cell of claim 8, wherein the nuclease comprises a zinc finger nuclease (ZFN), a TAL-effector domain nuclease (TALEN) or a CrispR/Cas system.

10. The RBC of claim 1, wherein the cell is type 0 negative.

11. The RBC precursor cell of claim 1, wherein the cell expresses fetal hemoglobin.

12. A method of degrading or detoxifying the effects of a toxin in a subject in need thereof, the method comprising administering a RBC or precursor RBC according to claim 1 to the subject in need thereof.

13. The method of claim 12, wherein RBC precursor cells are administered in a bone marrow transplant.

14. A method of providing a protein to a subject with a lysosomal storage disease, the method comprising administering a RBC or precursor RBC according to claim 1 to the subject in need thereof.