Signalling system
The present invention provides a chimeric antigen receptor (CAR) system comprising; (i) a receptor component comprising a antigen binding domain and a first binding domain; and (ii) a signalling component comprising a signalling domain and a second binding domain which binds the single domain binder of the first binding domain of the receptor component wherein either the first or second binding domains comprise a single domain binder, and wherein, binding of the first and second binding domains is disrupted by the presence of an agent, such that in the absence of the agent, the receptor component and the signalling component heterodimerize and binding of the antigen binding domain to antigen results in signalling through the signalling domain; whereas in the presence of the agent, the receptor component and the signalling component do not heterodimerize and binding of the antigen binding domain to antigen does not result in signalling through the signalling domain.
1. A chimeric antigen receptor (CAR) system comprising:
(i) a receptor component comprising an antigen binding domain, a transmembrane domain and first binding domain which comprises a single domain binder which binds an agent, and
(ii) an intracellular signalling component comprising a signalling domain and a second binding domain which binds the single domain binder of the first binding domain of the receptor component;
wherein binding of the first and second binding domains is disrupted by the presence of an agent, such that in the absence of the agent, the receptor component and the signalling component heterodimerize and binding of the antigen binding domain to antigen results in signalling through the signalling domain; whereas in the presence of the agent, the receptor component and the signalling component do not heterodimerize and binding of the antigen binding domain to antigen does not result in signalling through the signalling domain, and
wherein the agent is:
caffeine and
the single domain binder is a dAb comprising:
(a) CDR1 having the amino acid sequence TIYSMA (SEQ ID NO: 31), CDR2 having the amino acid sequence TVGWSSGITYYMDSVKG (SEQ ID NO: 32), and CDR3 having the amino acid sequence TRAYSVGYDY (SEQ ID NO: 33); or
(b) the amino acid sequence of SEQ ID NO: 11,
and the second binding domain comprises:
(SEQ ID NO. 12)
HTLNKPP,
(SEQ ID NO. 13)
DLSIGNH,
(SEQ ID NO. 14)
HDSPTAA,
(SEQ ID NO. 15)
YPDVPLA, or
(SEQ ID NO. 16)
ACTYLNSAKC.
2. A nucleic acid sequence encoding a CAR signalling system according to claim 1 , wherein the receptor component and signalling component are co-expressed with a self-cleaving peptide which is cleaved between the receptor component and the signalling component after translation of the components.
3. A vector comprising a nucleic acid sequence according to claim 2 .
4. A chimeric antigen receptor (CAR) system comprising;
(i) an intracellular signalling component comprising a signalling domain and a first binding domain comprises a single domain binder, and
(ii) a receptor component comprising an antigen binding domain, a transmembrane domain and second binding domain which binds the single domain binder of the intracellular signalling component;
wherein binding of the first and second binding domains is disrupted by the presence of an agent, such that in the absence of the agent, the receptor component and the signalling component heterodimerize and binding of the antigen binding domain to antigen results in signalling through the signalling domain; whereas in the presence of the agent, the receptor component and the signalling component do not heterodimerize and binding of the antigen binding domain to antigen does not result in signalling through the signalling domain and
wherein the agent is:
caffeine and
the single domain binder is a dAb comprising:
(a) CDR1 having the amino acid sequence TIYSMA (SEQ ID NO: 31), CDR2 having the amino acid sequence TVGWSSGITYYMDSVKG (SEQ ID NO: 32), and CDR3 having the amino acid sequence TRAYSVGYDY (SEQ ID NO: 33); or
(b) the amino acid sequence of SEQ ID NO: 11,
and the second binding domain comprises:
(SEQ ID NO. 12)
HTLNKPP,
(SEQ ID NO. 13)
DLSIGNH,
(SEQ ID NO. 14)
HDSPTAA,
(SEQ ID NO. 15)
YPDVPLA, or
(SEQ ID NO. 16)
ACTYLNSAKC.
5. A nucleic acid sequence encoding a CAR signalling system according to claim 4 , wherein the receptor component and signalling component are co-expressed with a self-cleaving peptide which is cleaved between the receptor component and the signalling component after translation of the components.
6. A vector comprising a nucleic acid sequence according to claim 5 .
7. A cell which expresses
(i) a receptor component comprising an antigen binding domain, a transmembrane domain and first binding domain which comprises a single domain binder which binds an agent, and
(ii) an intracellular signalling component comprising a signalling domain and a second binding domain which binds the single domain binder of the first binding domain of the receptor component;
wherein binding of the first and second binding domains is disrupted by the presence of an agent, such that in the absence of the agent, the receptor component and the signalling component heterodimerize and binding of the antigen binding domain to antigen results in signalling through the signalling domain; whereas in the presence of the agent, the receptor component and the signalling component do not heterodimerize and binding of the antigen binding domain to antigen does not result in signalling through the signalling domain and wherein the agent is:
caffeine and
the single domain binder is a dAb comprising:
(a) CDR1 having the amino acid sequence TIYSMA (SEQ ID NO: 31), CDR2 having the amino acid sequence TVGWSSGITYYMDSVKG (SEQ ID NO: 32), and CDR3 having the amino acid sequence TRAYSVGYDY (SEQ ID NO: 33); or
(b) the amino acid sequence of SEQ ID NO: 11,
and the second binding domain comprises:
(SEQ ID NO. 12)
HTLNKPP,
(SEQ ID NO. 13)
DLSIGNH,
(SEQ ID NO. 14)
HDSPTAA,
(SEQ ID NO. 15)
YPDVPLA, or
(SEQ ID NO. 16)
ACTYLNSAKC.
8. A pharmaceutical composition comprising a plurality of cells according to claim 7 .
9. A method for making a cell according to claim 7 , which comprises the step of introducing into the cell a nucleic acid sequence encoding or a vector encoding a CAR signalling system comprising:
(i) a receptor component comprising an antigen binding domain, a transmembrane domain and first binding domain which comprises a single domain binder which binds an agent, and
(ii) an intracellular signalling component comprising a signalling domain and a second binding domain which binds the single domain binder of the first binding domain of the receptor component;
wherein binding of the first and second binding domains is disrupted by the presence of an agent, such that in the absence of the agent, the receptor component and the signalling component heterodimerize and binding of the antigen binding domain to antigen results in signalling through the signalling domain; whereas in the presence of the agent, the receptor component and the signalling component do not heterodimerize and binding of the antigen binding domain to antigen does not result in signalling through the signalling domain, and
wherein the receptor component and signalling component are co-expressed with a self-cleaving peptide which is cleaved between the receptor component and the signalling component after translation of the components.
10. A cell which expresses
(i) an intracellular signalling component comprising a signalling domain and a first binding domain comprises a single domain binder, and
(ii) a receptor component comprising an antigen binding domain, a transmembrane domain and second binding domain which binds the single domain binder of the intracellular signalling component;
wherein binding of the first and second binding domains is disrupted by the presence of an agent, such that in the absence of the agent, the receptor component and the signalling component heterodimerize and binding of the antigen binding domain to antigen results in signalling through the signalling domain; whereas in the presence of the agent, the receptor component and the signalling component do not heterodimerize and binding of the antigen binding domain to antigen does not result in signalling through the signalling domain and
wherein the agent is:
caffeine and
the single domain binder is a dAb comprising:
(a) CDR1 having the amino acid sequence TIYSMA (SEQ ID NO: 31), CDR2 having the amino acid sequence TVGWSSGITYYMDSVKG (SEQ ID NO: 32), and CDR3 having the amino acid sequence TRAYSVGYDY (SEQ ID NO: 33); or
(b) the amino acid sequence of SEQ ID NO: 11,
and the second binding domain comprises:
(SEQ ID NO. 12)
HTLNKPP,
(SEQ ID NO. 13)
DLSIGNH,
(SEQ ID NO. 14)
HDSPTAA,
(SEQ ID NO. 15)
YPDVPLA, or
(SEQ ID NO. 16)
ACTYLNSAKC.
11. A pharmaceutical composition comprising a plurality of cells according to claim 10 .
12. A method for making a cell according to claim 10 , which comprises the step of introducing into the cell a nucleic acid sequence encoding or a vector encoding a CAR signalling system comprising:
(i) an intracellular signalling component comprising a signalling domain and a first binding domain comprises a single domain binder, and
(ii) a receptor component comprising an antigen binding domain, a transmembrane domain and second binding domain which binds the single domain binder of the intracellular signalling component;
wherein binding of the first and second binding domains is disrupted by the presence of an agent, such that in the absence of the agent, the receptor component and the signalling component heterodimerize and binding of the antigen binding domain to antigen results in signalling through the signalling domain; whereas in the presence of the agent, the receptor component and the signalling component do not heterodimerize and binding of the antigen binding domain to antigen does not result in signalling through the signalling domain, and
wherein the receptor component and signalling component are co-expressed with a self-cleaving peptide which is cleaved between the receptor component and the signalling component after translation of the components.