Genetically modified non-human animal with human or chimeric TIGIT
The present disclosure relates to the genetically modified non-human animals that express a human or chimeric TIGIT (e.g., humanized TIGIT), and methods of use thereof.
1. A method of determining effectiveness of an anti-human TIGIT (T cell immunoreceptor with Ig and ITIM domains) antibody for treating cancer, comprising:
administering the anti-human TIGIT antibody to a genetically-modified mouse, wherein the mouse has a tumor;
determining inhibitory effects of the anti-human TIGIT antibody on the tumor by measuring the size of the tumor in the mouse; and
comparing the size of the tumor in the mouse with the size of a tumor in a control mouse that is not treated with the anti-human TIGIT antibody,
wherein the genome of the mouse comprises a nucleic acid sequence encoding a chimeric TIGIT at an endogenous TIGIT gene locus, wherein the chimeric TIGIT comprises the amino acid sequence of SEQ ID NO: 34, and the nucleic acid sequence encoding the chimeric TIGIT is operably linked to an endogenous regulatory element at the endogenous TIGIT locus, wherein the mouse expresses the chimeric TIGIT and does not express endogenous TIGIT,
wherein the anti-human TIGIT antibody can bind to the expressed chimeric TIGIT protein and block the interaction of the chimeric TIGIT protein with a TIGIT ligand, thereby increasing immune response.
2. The method of claim 1 , wherein the mouse further comprises a sequence encoding an additional human or chimeric protein.
3. The method of claim 2 , wherein the additional human or chimeric protein is programmed cell death protein 1 (PD-1), CTLA-4, Lymphocyte Activating 3 (LAG-3), T-Cell Immunoglobulin And Mucin Domain-Containing Protein 3 (TIM-3), Programmed Cell Death 1 Ligand 1 (PD-L1), TNF Receptor Superfamily Member 9 (4-1BB), CD27, CD28, CD47, OX40, CD27, Glucocorticoid-Induced TNFR-Related Protein (GITR), or B And T Lymphocyte Associated (BTLA).
4. A method of determining effectiveness of an anti-human TIGIT antibody for treating cancer, comprising:
administering the anti-human TIGIT antibody to a genetically-modified mouse, wherein the mouse has a tumor;
determining inhibitory effects of the anti-human TIGIT antibody to the tumor by measuring the size of the tumor; and
comparing the size of the tumor in the mouse with the size of a tumor in a control mouse that is not treated with the anti-human TIGIT antibody,
wherein the genome of the mouse comprises a replacement of a portion of endogenous TIGIT exon 2 with a corresponding portion of human TIGIT exon 2, forming a chimeric TIGIT gene sequence at an endogenous TIGIT gene locus encoding a chimeric TIGIT, wherein a sequence within endogenous TIGIT exon 2 encoding at least 50 amino acid residues of the extracellular region of endogenous TIGIT protein is replaced with a sequence within human TIGIT exon 2 encoding at least 50 amino acid residues of the extracellular region of human TIGIT protein, wherein the chimeric TIGIT gene sequence is operably linked to an endogenous regulatory element, wherein the mouse expresses the chimeric TIGIT and does not express endogenous TIGIT, wherein the anti-human TIGIT antibody can bind to the expressed chimeric TIGIT protein and block the interaction of the chimeric TIGIT protein with a TIGIT ligand, thereby increasing immune response, wherein the chimeric TIGIT comprises the amino acid sequence of SEQ ID NO: 34.
5. The method of claim 1 , wherein the tumor comprises one or more human cancer cells that are injected into the mouse.
6. The method of claim 4 , wherein a sequence within endogenous TIGIT exon 2 encoding at least 80 amino acid residues of the extracellular region of endogenous TIGIT protein is replaced with a sequence within human TIGIT exon 2 encoding at least 80 amino acid residues of the extracellular region of human TIGIT protein.