Antibody/T-cell receptor chimeric constructs and uses thereof
The present application provides antibody-TCR chimeric constructs comprising an antibody moiety that specifically binds to a target antigen fused to a TCRM capable of recruiting at least one TCR-associated signaling module. Also provided are methods of making and using these constructs.
1. An antibody-T cell receptor (TCR) chimeric molecule (abTCR) that specifically binds to CD19, comprising:
a) a first polypeptide chain comprising a first antigen-binding domain comprising a V H antibody domain, a first constant antibody domain and a first T cell receptor domain (TCRD) comprising a first transmembrane domain of a first TCR subunit; and
b) a second polypeptide chain comprising a second antigen-binding domain comprising a V L antibody domain, a second constant antibody domain and a second TCRD comprising a second transmembrane domain of a second TCR subunit,
wherein the V H antibody domain of the first antigen-binding domain and the V L antibody domain of the second antigen-binding domain form an antigen-binding module that specifically binds to CD19,
wherein:
(i) the first TCR subunit is a TCR γ chain, and the second TCR subunit is a TCR δ chain; or
(ii) the first TCR subunit is a TCR δ chain, and the second TCR subunit is a TCR γ chain;
and wherein the first TCRD and the second TCRD form a T cell receptor module (TCRM) that is capable of recruiting at least one TCR-associated signaling module.
2. Nucleic acid(s) encoding the first and second polypeptide chains of the abTCR of claim 1 .
3. A complex comprising the abTCR of claim 1 and at least one TCR-associated signaling module selected from the group consisting of CD3δε, CD3γε, and ζζ.
4. An effector cell presenting on its surface the abTCR of claim 1 .
5. The effector cell of claim 4 , wherein the effector cell does not express the first TCR subunit and/or the second TCR subunit.
6. A method of killing a target cell presenting CD19, comprising contacting the target cell with the effector cell of claim 4 .
7. A pharmaceutical composition comprising the effector cell of claim 4 , and a pharmaceutically acceptable carrier.
8. A method of treating a CD19-associated disease in an individual in need thereof comprising administering to the individual an effective amount of the pharmaceutical composition of claim 7 .
9. The abTCR of claim 1 , wherein first and second constant antibody domains are each selected from the group consisting of a CH1, CH2, CH3, CH4 and CL antibody domain.
10. The effector cell of claim 4 , wherein the effector cell is a T cell.
11. The abTCR of claim 1 , wherein the first constant antibody domain is a CH1 antibody domain, and the second constant antibody domain is a CL antibody domain.
12. The abTCR of claim 1 , wherein:
(i) the V H antibody domain comprises heavy chain complementarity determining region (HC-CDR) 1, HC-CDR2 and HC-CDR3 of the amino acid sequence of SEQ ID NO: 45 and the V L antibody domain comprises light chain complementarity determining region (LC-CDR) 1, LC-CDR2 and LC-CDR3 of the amino acid sequence of SEQ ID NO: 46;
(ii) the V H antibody domain comprises HC-CDR1, HC-CDR2 and HC-CDR3 of the amino acid sequence of SEQ ID NO: 45, and the V L antibody domain comprises LC-CDR1, LC-CDR2 and LC-CDR3 of the amino acid sequence of SEQ ID NO: 57;
(iii) the V H antibody domain comprises HC-CDR1, HC-CDR2 and HC-CDR3 of the amino acid sequence of SEQ ID NO: 58, and the V L antibody domain comprises LC-CDR1, LC-CDR2 and LC-CDR3 of the amino acid sequence of SEQ ID NO: 57; or
(iv) the V H antibody domain comprises HC-CDR1, HC-CDR2 and HC-CDR3 of the amino acid sequence of SEQ ID NO: 59, and the V L antibody domain comprises LC-CDR1, LC-CDR2 and LC-CDR3 of the amino acid sequence of SEQ ID NO: 57.
13. The abTCR of claim 1 , wherein:
(i) the V H antibody domain comprises the amino acid sequence of SEQ ID NO: 45 and the V L antibody domain comprises the amino acid sequence of SEQ ID NO: 46;
(ii) the V H antibody domain comprises the amino acid sequence of SEQ ID NO: 45, and the V L antibody domain comprises the amino acid sequence of SEQ ID NO: 57;
(iii) the V H antibody domain comprises the amino acid sequence of SEQ ID NO: 58, and the V L antibody domain comprises the amino acid sequence of SEQ ID NO: 57; or
(iv) the V H antibody domain comprises the amino acid sequence of SEQ ID NO: 59, and the V L antibody domain comprises the amino acid sequence of SEQ ID NO: 57.
14. The abTCR of claim 1 , wherein the first polypeptide chain further comprises a first peptide linker between the first antigen-binding domain and the first TCRD and the second polypeptide chain further comprises a second peptide linker between the second antigen-binding domain and the second TCRD.
15. The abTCR of claim 1 , wherein the first TCRD further comprises a first connecting peptide or fragment thereof of a TCR subunit N-terminal to the first transmembrane domain and the second TCRD further comprises a second connecting peptide or fragment thereof of a TCR subunit N-terminal to the second transmembrane domain.
16. The abTCR of claim 1 , wherein the transmembrane domain of a TCR δ chain comprises the amino acid sequence of SEQ ID NO: 3 and the transmembrane domain of a TCR γ chain comprises the amino acid sequence of SEQ ID NO: 4.
17. The abTCR of claim 15 , wherein the connecting peptide of the TCRD comprising a transmembrane domain of a TCR δ chain comprises the amino acid sequence of SEQ ID NO: 7 and the connecting peptide of the TCRD comprising a transmembrane domain of a TCR γ chain comprises the amino acid sequence of SEQ ID NO: 8.
18. The abTCR of claim 15 , wherein the connecting peptide of the TCRD comprising a transmembrane domain of a TCR δ chain comprises the amino acid sequence of SEQ ID NO: 11 and the connecting peptide of the TCRD comprising a transmembrane domain of a TCR γ chain comprises the amino acid sequence of SEQ ID NO: 12.
19. The abTCR of claim 11 , wherein the CH1 antibody domain comprises the amino acid sequence of SEQ ID NO: 39 and the CL antibody domain comprises the amino acid sequence of SEQ ID NO: 41.
20. The abTCR of claim 1 , wherein:
(i) the first polypeptide comprises the amino acid sequence of SEQ ID NO: 42, and the second polypeptide comprises the amino acid sequence of SEQ ID NO: 54;
(ii) the first polypeptide comprises the amino acid sequence of SEQ ID NO: 42, and the second polypeptide comprises the amino acid sequence of SEQ ID NO: 43;
(iii) the first polypeptide comprises the amino acid sequence of SEQ ID NO: 55, and the second polypeptide comprises the amino acid sequence of SEQ ID NO: 54; or
(iv) the first polypeptide comprises the amino acid sequence of SEQ ID NO: 56, and the second polypeptide comprises the amino acid sequence of SEQ ID NO: 54.