Polypeptide
The present invention provides a chimeric polypeptide comprising: an antigen-binding domain which constitutively binds to an ectodomain of a first chain of a cytokine receptor; a transmembrane domain; and an endodomain from a second chain of the cytokine receptor which chimeric polypeptide, when expressed in a cell, binds to the endogenous first chain of the cytokine receptor causing constitutive cytokine signalling.
1. A chimeric polypeptide comprising:
an antigen-binding domain that constitutively binds to an ectodomain of a first chain of a cytokine receptor;
a transmembrane domain; and
an endodomain from a second chain of the cytokine receptor;
wherein the chimeric polypeptide, when expressed in a cell that expresses the cytokine receptor, binds to an endogenous first chain of the cytokine receptor expressed in the cell, causing constitutive cytokine signalling in the cell.
2. A chimeric polypeptide according to claim 1 , wherein:
the first chain of the cytokine receptor is a type I cytokine receptor γ-chain and the second chain of the cytokine receptor is type I cytokine receptor α-chain or β-chain,
or
the first chain of the cytokine receptor is a type I cytokine receptor α-chain or β-chain, and the second chain of the cytokine receptor is type I cytokine receptor γ-chain.
3. A chimeric polypeptide according to claim 2 , wherein the type I cytokine receptor α-chain or β-chain is from IL-2 receptor, IL-4 receptor, IL-7 receptor, IL-9 receptor, IL-13 receptor or IL-15 receptor.
4. A chimeric polypeptide according to claim 3 , wherein the type I cytokine receptor α-chain or β-chain is IL-7 receptor α-chain.
5. A chimeric polypeptide according to claim 1 , wherein the antigen-binding domain comprises a dAb or scFv that binds the ectodomain of the first chain of the cytokine receptor.
6. A constitutively-active cytokine receptor which comprises a chimeric polypeptide according to any preceding claim 1 in association with the first chain of the cytokine receptor.
7. A cell which comprises a constitutively-active cytokine receptor which comprises a chimeric polypeptide according to claim 1 in association with the endogenous first chain of the cytokine receptor.
8. A cell according to claim 7 , which also comprises a chimeric antigen receptor or an engineered T-cell receptor.
9. A nucleic acid comprising a nucleotide sequence encoding a chimeric polypeptide, the chimeric polypeptide comprising:
an antigen-binding domain that constitutively binds to an ectodomain of a first chain of a cytokine receptor;
a transmembrane domain; and
an endodomain from a second chain of the cytokine receptor;
wherein the chimeric polypeptide, when expressed in a cell that expresses the cytokine receptor, binds to an endogenous first chain of the cytokine receptor expressed in the cell, causing constitutive cytokine signaling in the cell.
10. A nucleic acid construct which comprises a first nucleic acid sequence encoding a chimeric polypeptide according to claim 1 ; and a second nucleic acid encoding a chimeric antigen receptor or an engineered T-cell receptor.
11. A vector comprising a nucleic acid according to claim 10 or a nucleic acid construct according to claim 10 .
12. A method for making a cell according to claim 7 , which comprises the step of introducing: a nucleic acid sequence into a cell, the nucleic acid sequence encoding a chimeric polypeptide comprising:
an antigen-binding domain that constitutively binds to an ectodomain of a first chain of a cytokine receptor;
a transmembrane domain; and
an endodomain from a second chain of the cytokine receptor;
wherein the chimeric polypeptide, when expressed in a cell that expresses the cytokine receptor, binds to an endogenous first chain of the cytokine receptor expressed in the cell, causing constitutive cytokine signalling in the cell.
13. A method for inducing constitutive cytokine signalling in a cell, which comprises the step of expressing a chimeric polypeptide according to claim 1 in the cell.
14. A method for blocking deleterious cytokine signalling in a cell, which comprises the step of expressing a chimeric polypeptide according to claim 1 in the cell, wherein the antigen binding domain constitutively binds to common gamma chain, and wherein one or more deleterious cytokine(s) signal(s) through a cytokine receptor which comprises common gamma chain.
15. A pharmaceutical composition comprising a plurality of cells according to claim 7 .
16. A method for treating a disease in a subject, which comprises the step of administering a pharmaceutical composition according to claim 15 to the subject.
17. A method for treating a disease in a subject which comprises the following steps:
(i) isolation of a cell-containing sample from the subject;
(ii) transduction or transfection of cells from the sample with the nucleic acid of claim; and
(iii) administering a pharmaceutical composition comprising the cells from (ii) to the subject.
18. A method according to claim 16 , wherein the disease is a cancer.
19. The chimeric polypeptide according to claim 1 , wherein the chimeric polypeptide further comprises a spacer that connects the antigen-binding domain with the transmembrane domain.
20. The chimeric polypeptide according to claim 19 , wherein the spacer comprises an IgG1 Fc region, an IgG1 hinge region, or a CD8 stalk.
21. The chimeric polypeptide according to claim 2 , wherein the first chain of the cytokine receptor is the type I cytokine receptor γ-chain.
22. The chimeric polypeptide according to claim 21 , wherein the antigen binding domain comprises an scFv which comprises light chain CDRs 1-3 of SEQ ID NO: 11-13, respectively, and heavy chain CDRs 1-3 shown in SEQ ID NO: 14-16, respectively.
23. The chimeric polypeptide according to claim 21 , wherein the transmembrane domain is derived from a cytokine receptor or from CD28.
24. The chimeric polypeptide according to claim 22 , wherein the transmembrane domain comprises SEQ ID NO: 7, 8, 9, or 10.
25. The chimeric polypeptide according to claim 23 , wherein the chimeric polypeptide further comprises a spacer that connects the antigen-binding domain with the transmembrane domain, wherein the spacer comprises an IgG1 Fc region, an IgG1 hinge region, or a CD8 stalk.
26. The chimeric polypeptide according to claim 24 , wherein the chimeric polypeptide further comprises a spacer that connects the antigen-binding domain with the transmembrane domain, wherein the spacer comprises SEQ ID NO: 4, 5, or 6.
27. The chimeric polypeptide according to claim 25 , wherein the endodomain of the chimeric polypeptide is from the IL-7 receptor α-chain.
28. The chimeric polypeptide according to claim 27 , wherein the endodomain of the chimeric polypeptide comprises SEQ ID NO: 3.
29. A chimeric polypeptide comprising:
an antigen-binding domain that comprises a dAb or scFv that binds to an ectodomain of a type I cytokine receptor y-chain;
a transmembrane domain derived from a cytokine receptor or from CD28;
a spacer derived from an IgG1 Fc region, an IgG1 hinge region, or a CD8 stalk;
an endodomain derived from a type I cytokine receptor α-chain or β-chain from IL-2 receptor, IL-4 receptor, IL-7 receptor, IL-9 receptor, IL-13 receptor or IL- 15 receptor;
wherein the chimeric polypeptide, when expressed in a cell that expresses the cytokine receptor, binds to an endogenous type I cytokine receptor γ-chain expressed in the cell, causing constitutive cytokine signalling in the cell.