Engineered immune cells with receptor cross-talk
The present disclosure provides engineered immune cells and methods for their creation and use. The immune cells comprise activating and blocking receptors, that exhibit cross-talk between the receptors.
1. An engineered immune cell comprising:
an activating receptor that triggers a cytotoxic signal that promotes a cytotoxic response of the engineered immune cell when the activating receptor binds a first ligand of a target cell; and
a blocking receptor that sends an interfering signal that inhibits the cytotoxic response of the engineered immune cell when the blocking receptor binds a second ligand of the target cell, each receptor comprising an extracellular binding domain, a hinge, a transmembrane domain, and an intracellular domain and said blocking receptor comprising a hinge that comprises a peptide of at least 24 contiguous amino acids of SEQ ID NO: 84, said peptide derived from a leukocyte immunoglobulin-like receptor subfamily B member 1 (LILRB1), wherein cross-talk between the activating receptor and the blocking receptor affects an activation threshold for the cytotoxic response.
2. The immune cell of claim 1 , wherein in the absence of the first and second ligands, the effect of the cross-talk on the activation threshold is minimized and/or reduced.
3. The immune cell of claim 1 , wherein the effect of the cross-talk on the activation threshold increases with proximity of the activating receptor to the blocking receptor.
4. The immune cell of claim 3 , wherein the activating receptor and blocking receptor are covalently linked together, or have physicochemical properties favoring interaction with one another such that the receptors are proximal to one another.
5. The immune cell of claim 1 , wherein when the blocking receptor binds to the second ligand, the cross-talk between the blocking and activating receptors causes the immune cell to exhibit reduced surface expression of the activating receptor.
6. The immune cell of claim 1 , wherein the immune cell comprises a plurality of the activating and blocking receptors, and when the immune cell contacts a target cell the plurality of the activating and blocking receptors diffuses into a region on the surface of the immune cell proximal to the target cell and forms a micro-cluster in which the effect of the cross-talk on the activation threshold is localized.
7. The immune cell of claim 1 , wherein the cross-talk between the activating receptor and the blocking receptor prevents the blocking receptor from binding to the second ligand until the activating receptor binds to the first ligand.
8. A method for treating cancer, the method comprising:
providing an engineered immune cell to a patient, wherein the engineered immune cell comprises an activating receptor and a blocking receptor, each expressed on a surface of the engineered immune cell, wherein:
the activating receptor triggers a cytotoxic signal that promotes a cytotoxic response of the engineered immune cell when the activating receptor binds a first ligand of a target cell; and
the blocking receptor sends an interfering signal that inhibits the cytotoxic response of the engineered immune cell when the blocking receptor binds a second ligand of the target cell, each receptor comprising an extracellular binding domain, a hinge, a transmembrane domain, and an intracellular domain and said blocking receptor comprising a hinge that comprises a peptide of at least 24 contiguous amino acids of SEQ ID NO: 84, said peptide derived from a leukocyte immunoglobulin-like receptor subfamily B member 1 (LILRB1), wherein cross-talk between the activating receptor and the blocking receptor affects an activation threshold for the cytotoxic response.
9. The method of claim 8 , wherein in the absence of the second ligand, the effect of the cross-talk on the activation threshold is minimized and/or reduced.
10. The method of claim 8 , wherein the effect of the cross-talk on the activation threshold increases with proximity of the activating receptor to the blocking receptor.
11. The method of claim 10 , wherein the activating receptor and blocking receptor are linked together or have physicochemical properties favoring interaction with one another, such that the receptors are proximal to one another.
12. The method of claim 8 , wherein when the blocking receptor binds to the second ligand, the cross-talk between the blocking and activating receptors causes the immune cell to exhibit reduced surface expression of the activating receptor.
13. The method of claim 8 , wherein the immune cell comprises a plurality of the activating and blocking receptors, and when the immune cell contacts a target cell the plurality of the activating and blocking receptors diffuses into a region on the surface of the immune cell proximal to the target cell and forms a micro-cluster in which the effect of the cross-talk on the activation threshold is localized.
14. The method of claim 8 , wherein the cross-talk between the activating receptor and the blocking receptor prevents the blocking receptor from binding to the second ligand until the activating receptor binds to the first ligand.
15. A method of producing an engineered immune cell, the method comprising:
determining an amount of cross-talk between an activating receptor and a blocking receptor for an engineered immune cell, wherein the amount of cross-talk between the activating receptor and the blocking receptor affects an activation threshold for the cytotoxic response; and
producing an engineered immune cell that expresses different concentrations of activating receptors and blocking receptors based on the determined amount of cross-talk between the activating receptor and the blocking receptor, each receptor comprising an extracellular binding domain, a hinge, a transmembrane domain, and an intracellular domain and said blocking receptor comprising a hinge that comprises a peptide of at least 24 contiguous amino acids of SEQ ID NO: 84, said peptide derived from a leukocyte immunoglobulin-like receptor subfamily B member 1 (LILRB1).
16. The method of claim 15 , wherein in the absence of cognate ligands for the activating and blocking receptors, the amount of the cross-talk is minimized and/or reduced.
17. The method of claim 15 , wherein producing an engineered immune cell that expresses different concentrations of activating receptors and blocking receptors is further based on a ratio of a quantity of an activating ligand to a quantity of a blocking ligand that are expressed in non-tumor cells of a sample.
18. The method of claim 17 , wherein the cross-talk between the activating receptor and the blocking receptor prevents the blocking receptor from binding to the blocking ligand until the activating receptor binds to the activating ligand.
19. The method of claim 15 , wherein an amount of the cross-talk between the activating receptor and blocking receptor increases with proximity of the activating receptor to the blocking receptor.
20. The method of claim 19 , wherein the activating receptor and blocking receptor are covalently linked, or have physicochemical properties favoring interaction with one another such that the receptors are proximal to one another.