Methods and compositions for engineering immunity

• US 10,639,383 B2
• Filed: 11/21/2016
• Issued: 05/05/2020
• Est. Priority Date: 11/23/2015
• Status: Active Grant

• Current Assignee: Sangamo Therapeutics, Inc.
• Original Assignee: Sangamo Therapeutics, Inc.
• Inventors: Holmes, Michael C., Riley, Brigit E.
• Primary Examiner(s): Bertoglio, Valarie E
• Application Number:
  US15/357,772
• Publication Number:
  US 20170196992A1
• Time in Patent Office: 1,261 Days
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2267/03 Animal model, e.g. for test or diseases
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• 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 48/0058 Nucleic acids adapted for tissue specific expression, e.g. having tissue specific promoters as part of a contruct
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 16/30 from tumour cells
• CPC Class Codes: C07K 16/32 against translation products of oncogenes
• CPC Class Codes: C07K 2317/622 Single chain antibody (scFv)
• CPC Class Codes: C07K 2317/626 Diabody or triabody
• CPC Class Codes: C07K 2319/31 fusions, other than Fc, for prolonged plasma life, e.g. albumin
• CPC Class Codes: C12N 15/62 DNA sequences coding for fusion proteins
• CPC Class Codes: C12N 15/86 Viral vectors
• CPC Class Codes: C12N 2750/14143 viral genome or elements thereof as genetic vector

Abstract

Nucleases and methods of using these nucleases for expressing an antibody from a safe harbor locus in a secretory tissue, and clones and animals derived therefrom.

Claims

1. A method of expressing, in a mammalian liver cell, an antibody, wherein the antibody binds to a protein expressed in the brain, wherein the protein is selected from the group consisting of a cancer antigen, a cellular receptor, a cytokine, a growth factor, a growth factor receptor, a kinase inhibitor, an integrin, α

-synuclein, an amyloid protein, and a complement protein, the method comprising,

administering to the liver cell (i) an adeno-associated viral (AAV) vector comprising a transgene encoding the antibody, wherein the antibody is a single chain fragment variable (ScFv), an intrabody or a diabody and (ii) one or more AAV vectors comprising sequences encoding a pair of zinc finger nucleases that cleave an endogenous albumin gene in the liver cell, such that the transgene is integrated into the liver cell and the liver cell produces and secretes the antibody such that the antibody is taken up by neurons in the brain.

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

3. The method of claim 1, wherein the transgene is a fusion protein comprising amino acids encoded by the transgene and by the endogenous albumin gene.

4. The method of claim 1, wherein the antibody is expressed in the serum and/or brain of the subject.

5. The method of claim 1, wherein the AAV vector is an AAV2 vector.

6. The method of claim 5, wherein the AAV2 vector is an AAV2/6 or AAV2/8 vector.

7. A genetically modified liver cell produced by the method of claim 1.