Genetically modified cells, tissues, and organs for treating disease

Patent metadata

Field	Value
Publication number	US-2019254265-A1
Application number	US-201916295887-A
Country	US
Kind code	A1
Filing date	Mar 7, 2019
Priority date	Dec 10, 2014
Publication date	Aug 22, 2019
Grant date	—

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

1 . (canceled)

2 . A genetically modified stem cell that comprises a disruption in a gene encoding NOD-like receptor family CARD domain containing 5 (NLRC5).

3 . The genetically modified stem cell of claim 2 , wherein said 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).

4 . The genetically modified stem cell of claim 2 , wherein said cell further comprises an exogenous polynucleotide that encodes a protein that comprises a human leukocyte antigen G (HLA-G) polypeptide sequence or a functional fragment thereof.

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

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

7 . The genetically modified stem cell of claim 2 , wherein said cell further comprises an exogenous polynucleotide that encodes a protein that comprises a beta 2 microglobulin (B2M) polypeptide sequence.

8 . The genetically modified stem cell of claim 2 , further comprising reduced protein expression of putative cytidine monophosphate-N-acetylneuraminic acid hydroxylase-like protein (CMAH), β1,4 N-acetylgalactosaminyltransferase (B4Ga1NT2), galactosyltransferase alpha 1,3 (GGTA1), MHC II transactivator (CIITA), or a combination thereof, as compared to a non-genetically modified counterpart stem cell.

9 . The genetically modified stem cell of claim 2 , further comprising a disruption in a gene encoding putative cytidine monophosphate-N-acetylneuraminic acid hydroxylase-like protein (CMAH), β1,4 N-acetylgalactosaminyltransferase (B4GalNT2), galactosyltransferase alpha 1,3 (GGTA1), β-2-microglobulin (B2M), or MEW II transactivator (CIITA).

10 . The genetically modified stem cell of claim 9 , comprising an exogenous polynucleotide encoding HLA-G inserted adjacent to the promoter of or inside one or more of: NOD-like receptor family CARD domain containing 5 (NLRC5), transporter associated with antigen processing 1 (TAP1), putative cytidine monophosphate-N-acetylneuraminic acid hydroxylase-like protein (CMAH), β1,4 N-acetylgalactosaminyltransferase (B4Ga1NT2), galactosyltransferase alpha 1,3 (GGTA1), β-2-microglobulin (B2M), and MEW II transactivator (CIITA).

11 . The genetically modified stem cell of claim 2 , wherein said disruption in said gene encoding NOD-like receptor family CARD domain containing 5 (NLRC5) is a nuclease mediated break in the stem cell DNA.

12 . The genetically modified stem cell of claim 2 , wherein the stem cell is an embryonic stem cell, pluripotent stem cell, or a differentiated stem cell.

13 . The genetically modified stem cell of claim 12 , wherein the stem cell is a human stem cell.

14 . The genetically modified stem cell of claim 12 , wherein the stem cell is from a member of the Laurasiatheria superorder.

15 . The genetically modified stem cell of claim 14 , wherein said member of the Laurasiatheria super order is an ungulate.

16 . The genetically modified stem cell of claim 15 , wherein said ungulate is a pig.

17 . A pharmaceutical composition that comprises a population of cells comprising the genetically modified stem cell of claim 2 , and a pharmaceutically acceptable excipient.

18 . The genetically modified stem cell of claim 12 , wherein said pluripotent stem cell is an induced pluripotent stem cell (iPSC), a clonal iPSC, or an iPS cell line cell.

19 . An insulin producing cell obtained by differentiating the genetically modified stem cell of claim 2 .

20 . A stem cell-derived, partially or fully differentiated cellular graft obtained by differentiating the genetically modified stem cell of claim 2 .

Assignees

• Univ Minnesota

Inventors

• Hering Bernhard J
• Burlak Christopher

Classifications

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• A61P3/10

  for hyperglycaemia, e.g. antidiabetics · CPC title

• A01K2267/025

  Animal producing cells or organs for transplantation · CPC title

• C07K16/2887

  against CD20 · CPC title

• A01K67/0275 Primary

  Genetically modified vertebrates, e.g. transgenic · CPC title