Chimeric antigen receptor (car) signalling system

Abstract

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)