Genetically modified mice and engraftment

Patent metadata

Field	Value
Publication number	US-11051499-B2
Application number	US-201916352674-A
Country	US
Kind code	B2
Filing date	Mar 13, 2019
Priority date	Oct 6, 2009
Publication date	Jul 6, 2021
Grant date	Jul 6, 2021

Abstract

A mouse with a humanization of the mIL-3 gene and the mGM-CSF gene, a knockout of a mRAG gene, and a knockout of a mII2rg subunit gene; and optionally a humanization of the TPO gene is described. A RAG/II2rg KO/hTPO knock-in mouse is described. A mouse engrafted with human hematopoietic stem cells (HSCs) that maintains a human immune cell (HIC) population derived from the HSCs and that is infectable by a human pathogen, e.g., S. typhi or M. tuberculosis is described. A mouse that models a human pathogen infection that is poorly modeled in mice is described, e.g., a mouse that models a human mycobacterial infection, wherein the mouse develops one or more granulomas comprising human immune cells. A mouse that comprises a human hematopoietic malignancy that originates from an early human hematopoietic cells is described, e.g., a myeloid leukemia or a myeloproliferative neoplasia.

First claim

We claim:

1. A genetically modified mouse, comprising a replacement of a mouse thrombopoietin (TPO) gene with a human TPO gene at both alleles of a mouse TPO gene locus, wherein the mouse:
does not express mouse TPO,
is immunocompromised for a mouse immune system,
is engrafted with human hematopoietic cells, and
is infected with a human pathogen.

2. The genetically modified mouse according to claim 1 , wherein the mouse comprises a human hemato-lymphoid system.

3. The genetically modified mouse according to claim 2 , wherein the human hemato-lymphoid system comprises human cells selected from the group consisting of hematopoietic stem cells, hematopoietic CD34+ cells, myeloid precursor cells, myeloid cells, dendritic cells, monocytes, granulocytes, neutrophils, mast cells, lymphocytes, and platelets.

4. The genetically modified mouse according to claim 1 , wherein the mouse is null for a RAG gene and null for the mouse interleukin 2 receptor gamma (IL-2Rγ) gene.

5. The genetically modified mouse according to claim 4 , wherein the mouse comprises a replacement of a mouse IL-3 gene with a human IL-3 gene at a mouse IL-3 gene locus.

6. The genetically modified mouse according to claim 5 , wherein the mouse comprises a replacement of a mouse GM-CSF gene with a human GM-CSF gene at a mouse GM-CSF gene locus.

7. The genetically modified mouse according to claim 1 , wherein the human pathogen is a mycobacterium.

8. The genetically modified mouse according to claim 7 , wherein the mycobacterium is Mycobacterium tuberculosis.

9. The genetically modified mouse according to claim 1 , wherein the human pathogen is Salmonella typhi.

10. A method of producing a genetically modified mouse comprising a human hemato-lymphoid system, the method comprising:
engrafting a population of cells that comprises human hematopoietic cells into an immunodeficient genetically modified mouse, wherein the genetically modified mouse comprises a replacement of a mouse thrombopoietin (TPO) gene with a human TPO gene at both alleles of a mouse TPO gene locus, wherein the mouse does not express mouse TPO, and
infecting the genetically modified mouse with a human pathogen.

11. The method according to claim 10 , wherein the human pathogen is a mycobacterium.

12. The method according to claim 11 , wherein the mycobacterium is Mycobacterium tuberculosis.

13. The method according to claim 10 , wherein the human pathogen is Salmonella typhi.

14. The method according to claim 10 , wherein the population of cells comprising human hematopoietic cells comprises a population of human umbilical cord blood cells or human fetal liver cells.

15. The method according to claim 10 , wherein said population of cells comprising human hematopoietic cells comprises human CD34+ cells.

16. The method according to claim 10 , wherein the human hemato-lymphoid system comprises human cells selected from the group consisting of hematopoietic stem cells, myeloid precursor cells, myeloid cells, dendritic cells, monocytes, granulocytes, neutrophils, mast cells, lymphocytes, and platelets.

17. The method according to claim 10 , further comprising irradiating the genetically modified mouse prior to the engrafting.

18. The method according to claim 10 , wherein the mouse is null for a RAG gene and null for the mouse interleukin 2 receptor gamma (IL-2Rγ) gene.

19. A mouse comprising a humanized cellular immune system, wherein the mouse is null for a RAG gene and null for the mouse interleukin 2 receptor gamma (IL-2Rγ) gene and is immunocompromised for a mouse immune system and comprises:
a replacement of a mouse thrombopoietin (TPO) gene with a human TPO gene at both alleles of a mouse TPO gene locus, wherein the mouse does not express mouse TPO;
a human hemato-lymphoid system; and
an infection with a human pathogen.

20. The mouse according to claim 19 , wherein the human pathogen is Mycobacterium tuberculosis or Salmonella typhi.

Assignees

• Regeneron Pharma
• Univ Yale
• Institute For Res In Biomedicine Irb

Inventors

• Stevens Sean
• Murphy Andrew J
• Flavell Richard
• Eynon Elizabeth
• Galan Jorge
• Willinger Tim
• Manz Markus
• Rongvaux Anthony
• Yancopoulos George D

Classifications

• Y02A50/30

  Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change · CPC title

• A01K2267/0331

  Animal model for proliferative diseases · CPC title

• C07K14/535

  Granulocyte CSF; Granulocyte-macrophage CSF · CPC title

• A01K2217/072

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

• A01K2227/105

  Murine · CPC title