Genetically modified non-human animal with human or chimeric CD3e
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.
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.