Genetically modified non-human animals and methods of use thereof

Patent metadata

Field	Value
Publication number	US-10433527-B2
Application number	US-201815980602-A
Country	US
Kind code	B2
Filing date	May 15, 2018
Priority date	Sep 7, 2012
Publication date	Oct 8, 2019
Grant date	Oct 8, 2019

Abstract

The invention relates generally to genetically modified non-human animals expressing human polypeptides and their methods of use.

First claim

The invention claimed is:

1. A method of transplanting human cancer cells, comprising:
transplanting human cancer cells into a genetically modified, immunodeficient mouse; and
transplanting human hematopoietic cells into the genetically modified, immunodeficient mouse,
wherein the genetically modified, immunodeficient mouse comprises in its genome a recombination activating gene 2 (Rag-2) gene knock-out, an IL2 receptor gamma chain (IL2 rg) gene knock-out, a replacement of a mouse M-CSF gene with a nucleic acid encoding a human M-CSF polypeptide at a mouse M-CSF gene locus, a replacement of a mouse IL-3 gene with a nucleic acid encoding a human IL-3 polypeptide at a mouse IL-3 gene locus, a replacement of a mouse GM-CSF gene with a nucleic acid encoding a human GM-CSF polypeptide at a mouse GM-CSF gene locus, an insertion of a nucleic acid encoding a human SIRPA polypeptide, and a replacement of a mouse TPO gene with a nucleic acid encoding a human TPO polypeptide at a mouse TPO gene locus, wherein each of the nucleic acids encoding the human M-CSF polypeptide, the human IL-3 polypeptide, the human GM-CSF polypeptide, the human SIRPA polypeptide, and the human TPO polypeptide is operably linked to a promoter, and wherein the mouse expresses the human M-CSF polypeptide, the human IL-3 polypeptide, the human GM-CSF polypeptide, the human SIRPA polypeptide, and the human TPO polypeptide.

2. The method of claim 1 , wherein the human cancer cells are primary human cancer cells isolated from a patient.

3. The method of claim 2 , wherein the human cancer cells and the human hematopoietic cells are isolated from the same patient.

4. The method of claim 1 , wherein the human cancer cells are from a cancer cell line.

5. The method of claim 1 , wherein the human cancer cells are selected from leukemia cells, breast cancer cells, lung cancer cells, and melanoma cells.

6. The method of claim 5 , wherein the human cancer cells are leukemia cells.

7. The method of claim 5 , wherein the human cancer cells are melanoma cells.

8. The method of claim 1 , wherein the human hematopoietic cells comprise CD34+cells.

9. The method of claim 1 , wherein the human SIRPA polypeptide is a biologically active fragment of a full-length human SIRPA polypeptide.

10. A method of human hematopoietic cell engraftment, comprising:
administering at least one human hematopoietic cell to a genetically modified, immunodeficient mouse, wherein the genetically modified, immunodeficient mouse comprises in its genome a recombination activating gene 2 (Rag-2) gene knock-out, an IL2 receptor gamma chain (IL2 rg) gene knock-out, a replacement of a mouse M-CSF gene with a nucleic acid encoding a human M-CSF polypeptide at a mouse M-CSF gene locus, a replacement of a mouse IL-3 gene with a nucleic acid encoding a human IL-3 polypeptide at a mouse IL-3 gene locus, a replacement of a mouse GM-CSF gene with a nucleic acid encoding a human GM-CSF polypeptide at a mouse GM-CSF gene locus, an insertion of a nucleic acid encoding a human SIRPA polypeptide, and a replacement of a mouse TPO gene with a nucleic acid encoding a human TPO polypeptide at a mouse TPO gene locus, wherein each of the nucleic acids encoding the human M-CSF polypeptide, the human IL-3 polypeptide, the human GM-CSF polypeptide, the human SIRPA polypeptide, and the human TPO polypeptide is operably linked to a promoter, and wherein the mouse expresses the human M-CSF polypeptide, the human IL-3 polypeptide, the human GM-CSF polypeptide, the human SIRPA polypeptide, and the human TPO polypeptide, and wherein the method does not comprise sub-lethally irradiating the genetically modified, immunodeficient mouse or treating the genetically modified, immunodeficient mouse with a radiomimetic drug prior to the administering.

11. The method of claim 10 , wherein the human SIRPA polypeptide is a biologically active fragment of a full-length human SIRPA polypeptide.

12. The method of claim 10 , wherein the human hematopoietic cells comprise CD34+cells.

Assignees

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

Inventors

• Flavell Richard
• Manz Markus
• Rongvaux Anthony
• Strowig Till
• Willinger Tim
• Murphy Andrew J
• Stevens Sean
• Yancopoulos George

Classifications

• A01K2207/15

  Humanized animals · CPC title

• A01K67/0271

  Chimeric vertebrates, e.g. comprising exogenous cells · CPC title

• A01K2207/12

  Animals modified by administration of exogenous cells · CPC title

• A01K2217/15

  Animals comprising multiple alterations of the genome, by transgenesis or homologous recombination, e.g. obtained by cross-breeding · CPC title

• A01K2267/0331

  Animal model for proliferative diseases · CPC title