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 chimeric antigen receptor (CAR) signalling system comprising;
(i) multiple receptor components comprising an extracellular antigen-binding domain, a transmembrane domain, and a intracellular first chemical inducer of dimerization (CID) binding domain (CBD1), wherein the antigen-binding domain of each receptor component binds to a different antigen; and
(ii) an intracellular signalling component comprising a signalling domain and a second CID binding domain (CBD2);
wherein CBD1 and CBD2 are capable of simultaneously binding to a CID; and
wherein, in the absence of CID, binding of the antigen-binding domain to antigen does not result in signalling through the signalling domain; whilst, in the presence of 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.
2 - 11 . (canceled)
12 . The CAR signalling system according to claim 1 , wherein the CBD1 of the multiple receptor components differ in binding to CID such that each antigen propagates different signalling strengths.
13 . The CAR signalling system according to claim 1 , wherein the signalling domain of the signalling component comprises a single endodomain selected from CD3 zeta endodomain, CD28 endodomain, 41 BB endodomain and OX40 endodomain.
14 . The CAR signalling system according to claim 1 , wherein the signalling domain of the signalling component comprises at least one of CD3 zeta endodomain, CD28 endodomain, 41 BB endodomain and OX40 endodomain.
15 - 18 . (canceled)
19 . A nucleic acid comprising nucleotide sequences encoding the CAR signaling system receptor component according to claim 1 .
20 . (canceled)
21 . A nucleic acid according to claim 19 , wherein the receptor component and signalling component are co-expressed by means of a self-cleaving peptide which is cleaved between the receptor component and the signalling component after translation.
22 . A nucleic acid according to claim 21 , wherein the multiple receptor components or multiple signalling components are co-expressed by means of a self-cleaving peptide which is cleaved between the receptor component(s) and the signalling component(s).
23 . A vector comprising a nucleic acid according to claim 18 .
24 . A retroviral vector or a lentiviral vector or a transposon comprising a vector according to claim 23 .
25 . A T cell or NK cell which expresses a CAR signaling system according to claim 1 .
26 . (canceled)
27 . A pharmaceutical composition comprising a plurality of T cells or NK cells according to claim 25 .
28 . (canceled)
29 . A method for treating and/or preventing a disease, which comprises the step of administering a pharmaceutical composition according to claim 27 to a subject.
30 . A method for treating and/or preventing a disease, which comprises the following steps:
(i) isolation of a T cell or NK containing sample from a subject;
(ii) transduction or transfection of the T or NK cells with a nucleic acid according to claim 18 or a vector comprising the nucleic acid sequence; and
(iii) administering the T cells or NK cells from (ii) to the subject.
31 . A method according to claim 29 , which further comprises the step of administering the CID to which the CBD1 and CBD2 simultaneously bind to the subject.
32 . A method for treating disease in a subject which subject comprises T cells or NK cells according to claim 25 , which comprises the step of administering the CID to which the CBD1 and CBD2 simultaneously bind to the subject.
33 . A method according to claim 29 , 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.
34 . A method according to claim 29 , wherein the disease is a cancer.
35 - 38 . (canceled)
39 . A method for modifying a T or NK cell, which comprises the step of introducing: a nucleic acid according to claim 21 or a vector comprising the nucleic add, into a T cell or NK cell.
40 . A method according to claim 39 , wherein the T or NK cell is from a sample isolated from a subject.
41 . A method for activating the CAR signalling system in a subject comprising a T or NK cell according to claim 25 , which method comprises the step of administering the CID to the subject.
42 . A method for reducing the activity of the CAR signalling system, in a subject comprising a T or NK cell according to claim 25 , which method comprises reducing or stopping administration of the CID to the subject.
43 . (canceled)