Genetically modified non-human animals and methods of use thereof

Patent metadata

Field	Value
Publication number	US-11576356-B2
Application number	US-202016782708-A
Country	US
Kind code	B2
Filing date	Feb 5, 2020
Priority date	Apr 13, 2015
Publication date	Feb 14, 2023
Grant date	Feb 14, 2023

Abstract

Genetically modified non-human animals expressing human SIRPα and human IL-15 from the non-human animal genome are provided. Also provided are methods for making non-human animals expressing human SIRPα and human IL-15 from the non-human animal genome, and methods for using non-human animals expressing human SIRPα and human IL-15 from the non-human animal genome. These animals and methods find many uses in the art, including, for example, in modeling human T cell and/or natural killer (NK) cell development and function, in modeling human pathogen infection of human T cells and/or NK cells, and in various in vivo screens.

First claim

That which is claimed is:

1. A genetically modified mouse, comprising:
a null mutation in the mouse SIRPα gene at the mouse SIRPα gene locus and a nucleic acid sequence encoding a human SIRPα protein incorporated into the genome of the genetically modified mouse and operably linked to an endogenous mouse SIRPα gene promoter at the mouse SIRPα gene locus; and
a null mutation in the mouse IL-15 gene at the mouse IL-15 gene locus and a nucleic acid sequence encoding a human IL-15 protein incorporated into the genome of the genetically modified mouse and is operably linked to an endogenous mouse IL-15 gene promoter at the mouse IL-15 gene locus,
wherein the genetically modified mouse expresses the human SIRPα protein and the human IL-15 protein, and
wherein the genetically modified mouse is immunodeficient and comprises a Rag2 gene knock-out and an IL2rg gene knock-out, a Rag2 gene knock-out, or an IL2rg gene knock-out.

2. The genetically modified mouse according to claim 1 , wherein the null mutation is a deletion of at least mouse SIRPα exons 2-4 and wherein the genetically modified mouse is heterozygous for the allele comprising the nucleic acid sequence encoding the human SIRPα protein.

3. The genetically modified mouse according to claim 1 , wherein the null mutation is a deletion of at least mouse SIRPα exons 2-4 and wherein the genetically modified mouse is homozygous for the allele comprising the nucleic acid sequence encoding the human SIRPα protein.

4. The genetically modified mouse according to claim 1 , wherein the human SIRPα protein is a functional fragment of a full length human SIRPα protein.

5. The genetically modified mouse according to claim 4 , wherein the functional fragment comprises an extracellular domain of human SIRPα.

6. The genetically modified mouse according to claim 1 , wherein the human IL-15 protein is a functional fragment of a full-length human IL-15 protein.

7. The genetically modified mouse according to claim 1 , wherein the null mutation is a deletion of at least mouse IL-15 exons 5-8, wherein the genetically modified mouse is heterozygous for the allele comprising the nucleic acid sequence encoding the human IL-15 protein.

8. The genetically modified mouse according to claim 1 , wherein the null mutation is a deletion of at least mouse IL-15 exons 5-8, wherein the genetically modified mouse is homozygous for the allele comprising the nucleic acid sequence that encodes the human IL-15 protein.

9. The genetically modified mouse according to claim 1 , wherein the genetically modified mouse comprises an engraftment of human hematopoietic cells.

10. The genetically modified mouse according to claim 9 , wherein the genetically modified mouse comprises an infection with a human pathogen.

11. The genetically modified mouse according to claim 10 , wherein the human pathogen activates, induces and/or targets T cells.

12. The genetically modified mouse according to claim 10 , wherein the human pathogen activates, induces and/or targets natural killer (NK) cells.

13. The genetically modified mouse according to claim 10 , wherein the human pathogen is a pathogen that infects human intestine or the human lung.

14. An animal engraftment model, comprising a genetically modified mouse comprising:
a null mutation in the mouse SIRPα gene at the mouse SIRPα gene locus and a nucleic acid sequence incorporated into the genome of the genetically modified mouse, which sequence encodes a human SIRPα protein and is operably linked to an endogenous mouse SIRPα gene promoter at the mouse SIRPα gene locus;
a null mutation in the mouse IL-15 gene at the mouse IL-15 gene locus and a nucleic acid sequence incorporated into the genome of the genetically modified mouse, which sequence encodes a human IL-15 protein and is operably linked to an endogenous mouse IL-15 gene promoter at the mouse IL-15 gene locus; and
an engraftment of human hematopoietic cells, wherein the genetically modified mouse (i) expresses the human SIRPα protein and the human IL-15 protein, and (ii) comprises human intraepithelial lymphocytes (IELs) in the lung or small intestine and Peyer's patches of the genetically modified mouse,
wherein the genetically modified mouse is immunodeficient and comprises a Rag2 gene knock-out and an IL2rg gene knock-out, a Rag2 gene knock-out, or an IL2rg gene knock-out.

15. The engraftment model according to claim 14 , wherein the null mutation is a deletion of at least mouse SIRPα exons 2-4, wherein the genetically modified mouse is heterozygous for the allele comprising the nucleic acid sequence that encodes the human SIRPα protein.

16. The engraftment model according to claim 14 , wherein the null mutation is a deletion of at least mouse SIRPα exons 2-4, wherein the genetically modified mouse is homozygous for the allele comprising the nucleic acid sequence that encodes the human SIRPα protein.

17. The engraftment model according to claim 14 , wherein the human SIRPα protein is a functional fragment of a full length human SIRPα protein and the human IL-15 protein is a functional fragment of a full-length human IL-15 protein.

18. The engraftment model according to claim 17 , wherein the functional fragment comprises an extracellular domain of human SIRPα.

19. The engraftment model according to claim 14 , wherein the null mutation is a deletion of at least mouse IL-15 exons 5-8, wherein the genetically modified mouse is heterozygous for the allele comprising the nucleic acid sequence that encodes the human IL-15 protein.

20. The engraftment model according to claim 14 , wherein the null mutation is a deletion of at least mouse IL-15 exons 5-8, wherein the genetically modified mouse is homozygous for the allele comprising the nucleic acid sequence that encodes the human IL-15 protein.

Assignees

• Regeneren Pharmaceuticals Inc
• Univ Yale
• Institute For Res In Biomedicine Irb
• Regeneron Pharma

Inventors

• Herndler-Brandstetter Dietmar
• Flavell Richard A
• Frleta Davor
• Gurer Cagan
• Manz Markus Gabriel
• Murphy Andrew J
• Palm Noah W
• Shan Liang
• Stevens Sean
• Strowig Till
• Yancopoulos George D
• De Zoete Marcel

Classifications

• A61P35/00

  Antineoplastic agents · CPC title

• C07K14/47

  from mammals · CPC title

• C07K14/5443

  IL-15 · CPC title

• A01K2207/12

  Animals modified by administration of exogenous cells · CPC title

• A01K2217/072

  maintaining or altering function, i.e. knock in · CPC title