Targeted disruption of the MHC cell receptor

• US 10,500,229 B2
• Filed: 12/15/2016
• Issued: 12/10/2019
• Est. Priority Date: 12/18/2015
• Status: Active Grant

• Current Assignee: Sangamo Therapeutics, Inc.
• Original Assignee: Sangamo Therapeutics, Inc.
• Inventors: Lee, Gary K., Paschon, David, Zhang, Lei
• Primary Examiner(s): Long, Scott
• Application Number:
  US15/380,723
• Publication Number:
  US 20170173080A1
• Time in Patent Office: 1,090 Days
• CPC Class Codes: A61K 35/00 Medicinal preparations containing materials or reaction products thereof with undetermined constitution
• CPC Class Codes: A61K 35/17 Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes when activated by a specific antigen A61K39/00
• 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: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 14/7051 T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 14/70521 CD28, CD152
• CPC Class Codes: C07K 14/70539 MHC-molecules, e.g. HLA-molecules
• CPC Class Codes: C07K 16/2803 against the immunoglobulin superfamily
• CPC Class Codes: C07K 2319/02 containing a signal sequence
• CPC Class Codes: C07K 2319/33 fusions for targeting to specific cell types, e.g. tissue specific targeting, targeting of a bacterial subspecies
• CPC Class Codes: C07K 2319/70 containing domain for protein-protein interaction
• CPC Class Codes: C12N 15/62 DNA sequences coding for fusion proteins
• CPC Class Codes: C12N 5/0636 T lymphocytes
• CPC Class Codes: C12N 9/22 Ribonucleases RNAses, DNAses catalytic nucleic acids C12N15/113

Abstract

Disclosed herein are methods and compositions for inactivating MHC genes, using zinc finger nucleases (ZFNs) comprising a zinc finger protein and a cleavage domain or cleavage half-domain in conditions able to preserve cell viability. Polynucleotides encoding ZFNs, vectors comprising polynucleotides encoding ZFNs and cells comprising polynucleotides encoding ZFNs and/or cells comprising ZFNs are also provided.

Claims

1. An isolated cell in which expression of a beta 2microglobulin (B2M) gene is partially or fully inactivated by an insertion and/or deletion within GGCCTTA, TCAAAT, TCAAATT, TTACTGA and/or AATTGAA of exon 1 or exon 2 of the B2M gene.

2. The cell of claim 1, wherein the modification is made by an exogenous fusion molecule comprising a DNA-binding domain that binds to a target site as shown in any of SEQ ID NO:

6-48 or 137 to 205 and a functional domain.

3. The cell of claim 1, further comprising an inactivated T-cell receptor gene, PD1 and/or CTLA4 gene.

4. The cell of claim 1, further comprising a transgene encoding a chimeric antigen receptor (CAR), a transgene encoding an Antibody-coupled T-cell Receptor (ACTR) and/or a transgene encoding an engineered TCR.

5. The cell of claim 1, wherein the cell is a lymphoid cell, a stem cell, or a progenitor cell.

6. The cell of claim 4, wherein the cell is a T-cell, an induced pluripotent stem cell (iPSC), an embryonic stem cell, a mesenchymal stem cell (MSC), or a hematopoietic stem cell (HSC).

7. A pharmaceutical composition comprising a cell according to claim 1.

8. A method of treating or preventing a cancer, the method comprising introducing a cell according to claim 1 to subject with cancer.