Chimeric antigen receptor (CAR) signalling system
The present invention relates to a chimeric antigen receptor (CAR) signalling system comprising; (i) a receptor component comprising an extracellular antigen-binding domain, a transmembrane domain and a intracellular first chemical inducer of dimerization binding domain 1 (CBD1); and (ii) an intracellular signalling component comprising a signalling domain and a second chemical inducer of dimerization binding domain 2 (CBD2); wherein CBD1 and CBD2 are capable of simultaneously binding to a chemical inducer of dimerization (CID); wherein, in the absence of the CID, binding of the antigen-binding component to antigen does not result in signalling through the signalling component; whilst, in the presence of the CID, the receptor component and the signalling component heterodimerize and binding of the antigen-binding domain to antigen results in signalling through the signalling domain.
1. A nucleic acid comprising a nucleotide sequence encoding a chimeric antigen receptor (CAR) signalling system, said signalling system comprising;
(i) a receptor component comprising an extracellular antigen-binding domain, a transmembrane domain and an intracellular first chemical inducer of dimerization (CID) binding domain (CBD1); and
(ii) a plurality of soluble intracellular signalling components that localise to the cytoplasm when expressed in a cell, each intracellular signalling component comprising a T cell receptor signalling domain and a second CID binding domain (CBD2), wherein the CBD2 of each intracellular signalling component recognises the same CID but the T cell receptor signalling domains comprise different endodomains;
wherein the nucleic acid sequence encodes a polypeptide that comprises the receptor component and the intracellular signalling components, joined by cleavage sites;
wherein CBD1 and CBD2 are capable of simultaneously binding to a CID; wherein, in the absence of CID, binding of the antigen-binding domain to antigen does not result in signalling through the T cell receptor signalling domain; whereas, in the presence of CID, the receptor component and the intracellular signalling component heterodimerize and binding of the antigen-binding domain to antigen results in T cell receptor signalling through the T cell receptor signalling domain.
2. The nucleic acid according to claim 1 , wherein the encoded receptor component and intracellular signalling components are joined by self-cleaving peptides which are cleaved between the receptor component and the intracellular signalling components after translation.
3. A vector comprising a nucleic acid according to claim 2 .
4. A T cell or NK cell which comprises the nucleic acid according to claim 2 , or which comprises a vector that comprises said nucleic acid.
5. A pharmaceutical composition comprising a plurality of T cells or NK cells according to claim 4 .
6. A method for treating and/or preventing a disease, which comprises the step of administering a pharmaceutical composition according to claim 5 to a subject.
7. A method according to claim 6 , which comprises the following steps:
(i) isolating a T cell or NK containing sample from a subject;
(ii) transducing or transfecting the T or NK cells with a nucleic acid sequence according to claim 2 or a vector comprising the nucleic acid sequence and
(iii) administering the T cells or NK cells from (ii) to the subject.
8. A method according to claim 6 , which further comprises the step of administering to the subject the CID to which the CBD1 and CBD2 are capable of simultaneously binding.
9. A method for treating and/or preventing a disease in a subject which subject comprises T cells or NK cells according to claim 4 , which comprises the step of administering the CID to which the CBD1 and CBD2 are capable of simultaneously binding.
10. A method according to claim 9 , which involves monitoring the progression of disease and/or monitoring toxic activity in the subject and adjusting the dose of the CID to provide acceptable levels of disease progression and/or toxic activity.
11. A method according to claim 6 , wherein the disease is a cancer.
12. A kit which comprises a nucleic acid according to claim 2 or a vector comprising the nucleic acid, and the CID to which the CBD1 and CBD2 are capable of simultaneously binding.
13. A kit which comprises a T or NK cell according to claim 4 and the CID to which the CBD1 and CBD2 are capable of simultaneously binding.
14. A method for making a T or NK cell that comprises a chimeric antigen receptor (CAR) signalling system, which comprises the step of introducing: a nucleic acid according to claim 2 , or a vector comprising the nucleic acid, into a T cell or NK cell.
15. A method according to claim 14 , wherein the T or NK cell is from a sample isolated from a subject.
16. A method for activating a CAR signalling system, in a subject comprising a T or NK cell according to claim 4 , which method comprises the step of administering the CID to the subject.
17. A method for reducing the activity of a CAR signalling system, in a subject comprising a T or NK cell according to claim 4 , which method comprises reducing or stopping administration of the CID to the subject.
18. A method for inducing dimerization in vivo between a receptor component and an intracellular signalling component in a subject comprising a T or NK cell according to claim 4 , which comprises the step of administering a chemical inducer of dimerization (CID) to the subject.
19. A vector comprising a nucleic acid according to claim 1 .
20. A T cell or NK cell which comprises the vector according to claim 19 .
21. A T cell or NK cell that expresses a chimeric antigen receptor (CAR) signalling system, said signalling system comprising;
(i) a receptor component comprising an extracellular antigen-binding domain, a transmembrane domain, and a intracellular first chemical inducer of dimerization (CID) binding domain (CBD1); and
(ii) a plurality of soluble intracellular signalling components that localise to the cytoplasm when expressed in the cell, each intracellular signalling component comprising a T cell receptor signalling domain and a second CID binding domain (CBD2);
wherein CBD1 and CBD2 are capable of simultaneously binding to a CID; wherein, in the absence of CID, binding of the antigen-binding domain to antigen does not result in signalling through the T cell receptor signalling domain; whereas, in the presence of CID, the receptor component and the intracellular signalling component heterodimerize and binding of the antigen-binding domain to antigen results in T cell receptor signalling through the T cell receptor signalling domain,
wherein the CBD2 of each of the plurality of intracellular signalling components recognises the same CID but the T cell receptor signalling domains of the plurality of intracellular signalling components comprise different endodomains.
22. A pharmaceutical composition comprising a plurality of T cells or NK cells according to claim 21 .
23. The nucleic acid according to claim 1 , wherein the CAR signalling system has a single receptor component.
24. The T cell or NK cell according to claim 21 , wherein the CAR signalling system has a single receptor component.