Constitutively active chimeric cytokine receptors
Provided herein are constitutively active chimeric cytokine receptors (CACCRs). When present on chimeric antigen receptor (CAR)-bearing immune cells, such CACCRs allow for increased immune cell activation, proliferation, persistence, and/or potency. Also provided are methods of making and using the CACCRs described herein.
1. A constitutively active chimeric cytokine receptor (CACCR) composed of two monomers, each monomer comprising:
a. a transmembrane domain;
b. a Janus Kinase (JAK)-binding domain; and
c. a recruiting domain,
wherein the monomers are constitutively dimerized and wherein the transmembrane domain and JAK-binding domain comprises amino acids 478-582 of SEQ ID NO: 6 and comprises amino acid substitutions S505N and W515K.
2. The CACCR of claim 1 , wherein the JAK-binding domain comprises a JAK1-binding domain, a JAK2-binding domain, a JAK3-binding domain or a TYK2-binding domain.
3. The CACCR of claim 1 , wherein the recruiting domain comprises a STAT-recruiting domain from at least one receptor.
4. The CACCR of claim 3 , wherein the STAT-recruiting domain is from IL2Rb.
5. The CACCR of claim 1 , wherein amino acids 478-582 of SEQ ID NO: 6 further comprise an amino acid substitution H499L.
6. The CACCR of claim 5 , wherein the transmembrane domain comprises SEQ ID NO: 12.
7. The CACCR of claim 1 , wherein amino acids 478-582 of SEQ ID NO: 6 further comprise an amino acid substitution G509N.
8. The CACCR of claim 1 , wherein amino acids 478-582 of SEQ ID NO: 6 further comprise amino acid substitutions H499L and G509N.
9. The CACCR of claim 1 , wherein the transmembrane domain comprises SEQ ID NO: 13.
10. The CACCR of claim 1 , wherein the monomers are identical.
11. The CACCR of claim 1 , wherein the monomers are different.
12. The CACCR of claim 1 , wherein the recruiting domain comprises a STAT-recruiting domain from a cytokine receptor.
13. The CACCR of claim 1 , wherein the recruiting domain comprises a STAT-recruiting domain from a receptor selected from BLNK, IL2RG, EGFR, EpoR, GHR, IFNAR1, IFNAR2, IFNAR1/2, IFNLR1, IL10R1, IL12Rb1, IL12Rb2, IL21R, IL2Rb, IL2small, IL7R, IL7Ra, IL9R, IL15R, and IL21R.
14. The CACCR of claim 1 , wherein the recruiting domain comprises the amino acid sequence of one or more of the receptor sequences selected from SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 52, SEQ ID NO: 53, SEQ ID NO: 54, SEQ ID NO: 55, SEQ ID NO: 56, SEQ ID NO: 57, SEQ ID NO: 58, SEQ ID NO: 59, SEQ ID NO: 60, SEQ ID NO: 61, SEQ ID NO: 62, SEQ ID NO: 63, SEQ ID NO: 64, SEQ ID NO: 65, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 73, SEQ ID NO: 74, SEQ ID NO: 75, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 80, SEQ ID NO: 81, SEQ ID NO: 82, SEQ ID NO: 83, SEQ ID NO: 84, SEQ ID NO: 85, SEQ ID NO: 86, SEQ ID NO: 87, SEQ ID NO: 88, SEQ ID NO: 106, and SEQ ID NO: 143.
15. The CACCR of claim 1 , wherein the recruiting domain comprises a STAT-recruiting domain from IL7Ra.
16. The CACCR of claim 1 , wherein the recruiting domain comprises SEQ ID NO: 46.
17. The CACCR of claim 1 , wherein the recruiting domain comprises a STAT-recruiting domain from IL2Rb.
18. The CACCR of claim 1 , wherein the recruiting domain comprises (i) amino acid residues 1 to 41 of SEQ ID NO: 73, (ii) SEQ ID NO: 74, (iii) SEQ ID NO: 77, or (iv) SEQ ID NO: 78.
19. The CACCR of claim 1 , wherein the recruiting domain comprises a STAT-recruiting domain from IL12Rb2.
20. The CACCR of claim 1 , wherein the recruiting domain comprises SEQ ID NO: 86 or 67.
21. The CACCR of claim 1 , wherein the recruiting domain comprises STAT-recruiting domains from two receptors.
22. The CACCR of claim 1 , wherein the recruiting domain comprises STAT-recruiting domains from two cytokine receptors.
23. The CACCR of claim 22 , wherein the two cytokine receptors are selected from the group consisting of IL7Ra, IL2Rb, and IL12Rb2.
24. The CACCR of claim 1 , wherein the recruiting domain comprises SEQ ID NO: 77.
25. The CACCR of claim 1 , wherein the recruiting domain comprises SEQ ID NO: 78.
26. A polynucleotide encoding the CACCR of claim 1 .
27. An expression vector comprising the polynucleotide of claim 26 .
28. The expression vector of claim 27 further comprising a polynucleotide expressing a chimeric antigen receptor (CAR).
29. The expression vector of claim 27 , wherein the vector is a lentiviral vector.
30. An engineered immune cell comprising the expression vector of claim 27 .
31. The engineered immune cell of claim 30 , wherein the immune cell is a T-cell.
32. An engineered immune cell comprising a chimeric antigen receptor (CAR) and at least one CACCR of claim 1 .
33. The engineered immune cell of claim 32 , wherein the CAR and the CACCR are expressed in stoichiometrically equal amounts.
34. The engineered immune cell of claim 33 , wherein the immune cell is a T-cell.
35. A method of preparing an engineered immune cell, the method comprising introducing the polynucleotide of claim 26 into an immune cell.
36. A pharmaceutical composition comprising the engineered immune cell of claim 30 .
37. A kit comprising the engineered immune cell of claim 30 .
38. A method of treating a cancer in a subject, comprising administering to the subject a therapeutically effective amount of the engineered immune cell of claim 30 .
39. The method of claim 38 , wherein the cancer comprises a solid tumor or a liquid tumor.