Genetically modified non-human animal with human or chimeric CD3e

Patent metadata

Field	Value
Publication number	US-12016315-B2
Application number	US-202117168603-A
Country	US
Kind code	B2
Filing date	Feb 5, 2021
Priority date	Dec 12, 2017
Publication date	Jun 25, 2024
Grant date	Jun 25, 2024

Abstract

The present disclosure relates to genetically modified non-human animals that express a human or chimeric (e.g., humanized) CD3e (T-cell surface glycoprotein CD3 epsilon chain), and methods of use thereof.

First claim

What is claimed is:

1. A method of determining effectiveness of a therapeutic agent targeting CD3 for treating cancer, comprising:
administering the therapeutic agent targeting CD3 to a genetically-modified, non-human mammal whose genome comprises a replacement of a nucleic acid sequence encoding all or a portion of the extracellular region of an endogenous CD3e with a nucleic acid sequence encoding the corresponding region of a human CD3e, wherein the replacement results in a sequence encoding a chimeric CD3e comprising an amino acid sequence that is at least 95% identical to amino acids 1-126 of SEQ ID NO: 7, wherein the chimeric CD3e associates with endogenous CD3γ and endogenous CD3δ, forming a functional CD3 complex on T cells in the mammal, wherein the non-human mammal detectably expresses the chimeric CD3e on the surface of one or more T cells, wherein the mammal has a cancer; and
determining the inhibitory effects of the therapeutic agent to the cancer.

2. The method of claim 1 , wherein the cancer comprises one or more cancer cells that are injected into the mammal.

3. The method of claim 1 , wherein the cancer cells are melanoma cells, pancreatic carcinoma cells, mesothelioma cells, or solid tumor cells.

4. A method of determining effects of a therapeutic agent targeting CD3 on immune response, comprising:
administering the therapeutic agent targeting CD3 to a genetically-modified, non-human mammal whose genome comprises a replacement of a nucleic acid sequence encoding all or a portion of the extracellular region of an endogenous CD3e with a nucleic acid sequence encoding the corresponding region of a human CD3e, wherein the replacement results in a sequence encoding a chimeric CD3e comprising an amino acid sequence that is at least 95% identical to amino acids 1-126 of SEQ ID NO: 7, wherein the chimeric CD3e associates with endogenous CD3γ and endogenous CD3δ, forming a functional CD3 complex on T cells in the mammal, wherein the non-human mammal detectably expresses the chimeric CD3e on the surface of one or more T cells; and
determining the effects of the therapeutic agent targeting CD3.

5. The method of claim 4 , wherein the therapeutic agent targeting CD3 inhibits an immune response.

6. The method of claim 4 , wherein the therapeutic agent targeting CD3 stimulates an immune response.

7. The method of claim 4 , wherein the therapeutic agent targeting CD3 activates T cells.

8. The method of claim 4 , wherein the effects are determined by measuring the percentage of T cells in peripheral blood.

9. The method of claim 4 , wherein the sequence encoding the chimeric CD3e is operably linked to an endogenous regulatory element at an endogenous CD3e gene locus.

10. The method of claim 4 , wherein the chimeric CD3e comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 10.

11. The method of claim 4 , wherein the chimeric CD3e comprises an amino acid sequence that is identical to SEQ ID NO: 10.

12. The method of claim 4 , wherein the mammal is a rodent or a mouse.

13. The method of claim 4 , wherein the therapeutic agent targeting CD3 is an anti-human CD3e antibody or antigen-binding fragment thereof.

14. The method of claim 4 , wherein the mammal further comprises a sequence encoding an additional human or chimeric protein.

15. The method of claim 5 , wherein the additional human or chimeric protein is programmed cell death protein 1 (PD-1), cytotoxic T-lymphocyte-associated protein 4 (CTLA-4), Lymphocyte Activating 3 (LAG-3), B And T Lymphocyte Associated (BTLA), Programmed Cell Death 1 Ligand 1 (PD-L1), CD27, CD28, CD47, CD137, CD154, T-Cell Immunoreceptor With Ig And ITIM Domains (TIGIT), T-cell Immunoglobulin and Mucin-Domain Containing-3 (TIM-3), Glucocorticoid-Induced TNFR-Related Protein (GITR), TNF Receptor Superfamily Member 4 (OX40), CD3δ, CD3γ, CD40, or CD278.

16. A method of determining effects of a therapeutic agent targeting CD3 on immune response, comprising:
administering the therapeutic agent targeting CD3 to a genetically-modified, non-human mammal, whose genome comprises a replacement of a nucleic acid sequence encoding an endogenous CD3e with a nucleic acid sequence encoding a human CD3e, wherein the replacement results in a sequence encoding the human CD3e comprising an amino acid sequence that is at least 95% identical to amino acids 1-126 of SEQ ID NO: 7, wherein the human CD3e associates with endogenous CD3γ and endogenous CD3δ, forming a functional CD3 complex on T cells in the mammal, wherein the non-human mammal detectably expresses the human CD3e on the surface of one or more T cells; and
determining the effects of the therapeutic agent targeting CD3.

17. The method of claim 16 , wherein the human CD3e comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 7.

18. The method of claim 16 , wherein the mammal is a rodent or a mouse.

19. The method of claim 16 , wherein the therapeutic agent targeting CD3 is an anti-human CD3e antibody or antigen-binding fragment thereof.

Assignees

• Biocytogen Pharmaceuticals Beijing Co Ltd
• Biocytogen Jiangsu Co Ltd

Inventors

• Shen Yuelei
• Guo Yanan
• Huang Rui
• Zhou Xiaofei
• Bai Yang
• Yao Jiawei
• Guo Chaoshe

Classifications

• A61K35/51

  Umbilical cord; Umbilical cord blood; Umbilical stem cells · CPC title

• A61K49/0008

  Screening agents using (non-human) animal models or transgenic animal models or chimeric hosts, e.g. Alzheimer disease animal model, transgenic model for heart failure · CPC title

• A01K2267/0331

  Animal model for proliferative diseases · CPC title

• A01K2227/105

  Murine · CPC title

• A01K2207/15

  Humanized animals · CPC title