Genetically modified cells, tissues, and organs for treating disease

Patent metadata

Field	Value
Publication number	US-10993419-B2
Application number	US-201916295887-A
Country	US
Kind code	B2
Filing date	Mar 7, 2019
Priority date	Dec 10, 2014
Publication date	May 4, 2021
Grant date	May 4, 2021

Abstract

Genetically modified cells, tissues, and organs for treating or preventing diseases are disclosed. Also disclosed are methods of making the genetically modified cells and non-human animals.

First claim

What is claimed:

1. A genetically modified human stem cell that comprises a disruption in a gene encoding NOD-like receptor family CARD domain containing 5 (NLRC5), wherein said disruption in said gene encoding NLRC5 is a nuclease mediated double stranded break at a target sequence in said gene, and wherein said target sequence is located within 20 nucleotides of a protospacer-adjacent motif (PAM) sequence.

2. The genetically modified human stem cell of claim 1 , wherein said genetically modified human stem cell has a reduced protein expression of NOD-like receptor family CARD domain containing 5 (NLRC5) as compared to a corresponding cell that does not have a disruption in said gene encoding NOD-like receptor family CARD domain containing 5 (NLRC5).

3. A genetically modified stem cell that comprises:
a disruption in a gene encoding NOD-like receptor family CARD domain containing 5 (NLRC5); and
an exogenous polynucleotide that encodes a protein that comprises a human leukocyte antigen G (HLA-G) polypeptide sequence or a functional fragment thereof.

4. The genetically modified stem cell of claim 3 , wherein said HLA-G is HLA-G1, HLA-G2, HLA-G3, HLA-G4, HLA-G5, HLA-G6, HLA-G7, or a functional fragment thereof.

5. The genetically modified stem cell of claim 4 , wherein said HLA-G is HLA-G1 or a functional fragment thereof.

6. A genetically modified stem cell that comprises;
a disruption in a gene encoding NOD-like receptor family CARD domain containing 5 (NLRC5); and an exogenous polynucleotide that encodes a protein that comprises a beta 2 microglobulin (B2M) polypeptide sequence.

7. The genetically modified human stem cell of claim 1 , that is an embryonic stem cell, or a pluripotent stem cell.

8. A pharmaceutical composition that comprises a plurality of said genetically modified human stem cells of claim 7 , or cells differentiated therefrom,
and a pharmaceutically acceptable excipient.

9. The genetically modified human stem cell of claim 7 , wherein said pluripotent stem cell is an induced pluripotent stem cell (iPSC), a clonal iPSC, or an iPS cell line cell.

10. A genetically modified stem cell comprising;
a disruption in a gene encoding β-2-microglobulin (B2M); and
an exogenous polynucleotide that encodes a programmed death-ligand 1 (PD-L1) or a functional fragment thereof.

11. An insulin producing cell obtained by differentiating the genetically modified stem cell of claim 10 .

12. The insulin producing cell of claim 11 , wherein the cell is non-cancerous.

Assignees

• Univ Minnesota

Inventors

• Hering Bernhard J
• Burlak Christopher

Classifications

• A61K35/15

  Cells of the myeloid line, e.g. granulocytes, basophils, eosinophils, neutrophils, leucocytes, monocytes, macrophages or mast cells; Myeloid precursor cells; Antigen-presenting cells, e.g. dendritic cells (presenting a specific antigen A61K39/00; therapeutic combinations of antibodies, or fragments thereof, and blood-derived cells A61K39/00) · CPC title

• C07K16/2833

  against MHC-molecules, e.g. HLA-molecules · CPC title

• A61K31/436

  the heterocyclic ring system containing a six-membered ring having oxygen as a ring hetero atom, e.g. rapamycin · CPC title

• A61K35/28

  Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells · CPC title

• A01K2267/025

  Animal producing cells or organs for transplantation · CPC title