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. A method of killing a target cell presenting a target antigen, comprising contacting the target cell with an effector αβ T cell comprising an antibody-T cell receptor (TCR) chimeric molecule (abTCR) that specifically binds to the target antigen, wherein the abTCR comprises:
a) a first polypeptide chain comprising a first antigen-binding domain comprising a V H antibody domain and a C H 1 antibody domain, and a first T cell receptor domain (TCRD) comprising a first transmembrane domain of a first TCR subunit, wherein the first polypeptide chain lacks variable and constant domains of the first TCR subunit; and
b) a second polypeptide chain comprising a second antigen-binding domain comprising a V L antibody domain and a C L antibody domain, and a second TCRD comprising a second transmembrane domain of a second TCR subunit, wherein the second polypeptide chain lacks variable and constant domains of the second TCR subunit,
wherein the V H domain of the first antigen-binding domain pairs with the V L domain of the second antigen-binding domain to form an antigen-binding module that specifically binds to the target antigen,
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, and
wherein the first TCR subunit is a TCR γ chain and the second TCR subunit is a TCR δ chain, or the first TCR subunit is a TCR δ chain and the second TCR subunit is a TCR γ chain.
2. The method of claim 1 , wherein the first TCR subunit is a TCR γ chain, and the second TCR subunit is a TCR δ chain.
3. The method of claim 1 , wherein the first TCR subunit is a TCR δ chain, and the second TCR subunit is a TCR γ chain.
4. The method of claim 1 , wherein the contacting is in vivo.
5. The method of claim 1 , wherein the contacting is in vitro.
6. A method of treating a target antigen-associated disease in an individual in need thereof comprising administering to the individual an effective amount of a composition comprising an effector αβ T cell comprising an abTCR that specifically binds to the target antigen, wherein the abTCR comprises:
a) a first polypeptide chain comprising a first antigen-binding domain comprising a V H antibody domain and a C H 1 antibody domain, and a first TCRD comprising a first transmembrane domain of a first TCR subunit, wherein the first polypeptide chain lacks variable and constant domains of the first TCR subunit; and
b) a second polypeptide chain comprising a second antigen-binding domain comprising a V L antibody domain and a C L antibody domain, and a second TCRD comprising a second transmembrane domain of a second TCR subunit, wherein the second polypeptide chain lacks variable and constant domains of the second TCR subunit,
wherein the V H domain of the first antigen-binding domain pairs with the V L domain of the second antigen-binding domain to form an antigen-binding module that specifically binds to the target antigen,
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, and
wherein the first TCR subunit is a TCR γ chain and the second TCR subunit is a TCR δ chain, or the first TCR subunit is a TCR δ chain and the second TCR subunit is a TCR γ chain.
7. The method of claim 6 , wherein the first TCR subunit is a TCR γ chain, and the second TCR subunit is a TCR δ chain.
8. The method of claim 6 , wherein the first TCR subunit is a TCR δ chain, and the second TCR subunit is a TCR γ chain.
9. The method of claim 6 , wherein the target antigen-associated disease is cancer.
10. The method of claim 6 , wherein the target antigen-associated disease is viral infection.
11. The method 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.
12. The method of claim 11 , wherein the TCRM comprises a disulfide bond between a residue in the first connecting peptide and a residue in the second connecting peptide.
13. The method of claim 11 , wherein:
(i) the first TCR subunit is a TCR δ chain, and the second TCR subunit is a TCR γ chain, wherein the first connecting peptide comprises the amino acid sequence of SEQ ID NO: 7, and the second connecting peptide comprises the amino acid sequence of SEQ ID NO: 8; or
(ii) the first TCR subunit is a TCR γ chain, and the second TCR subunit is a TCR δ chain, wherein the first connecting peptide comprises the amino acid sequence of SEQ ID NO: 8, and the second connecting peptide comprises the amino acid sequence of SEQ ID NO: 7.
14. The method of claim 11 , wherein:
(i) the first TCR subunit is a TCR δ chain, and the second TCR subunit is a TCR γ chain, wherein the first connecting peptide comprises the amino acid sequence of SEQ ID NO: 11, and the second connecting peptide comprises the amino acid sequence of SEQ ID NO: 12; or
(ii) the first TCR subunit is a TCR γ chain, and the second TCR subunit is a TCR δ chain, wherein the first connecting peptide comprises the amino acid sequence of SEQ ID NO: 12, and the second connecting peptide comprises the amino acid sequence of SEQ ID NO: 11.
15. The method of claim 6 , 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 method of claim 15 , wherein the TCRM comprises a disulfide bond between a residue in the first connecting peptide and a residue in the second connecting peptide.
17. The method of claim 15 , wherein:
(i) the first TCR subunit is a TCR δ chain, and the second TCR subunit is a TCR γ chain, wherein the first connecting peptide comprises the amino acid sequence of SEQ ID NO: 7, and the second connecting peptide comprises the amino acid sequence of SEQ ID NO: 8; or
(ii) the first TCR subunit is a TCR γ chain, and the second TCR subunit is a TCR δ chain, wherein the first connecting peptide comprises the amino acid sequence of SEQ ID NO: 8, and the second connecting peptide comprises the amino acid sequence of SEQ ID NO: 7.
18. The method of claim 15 , wherein:
(i) the first TCR subunit is a TCR δ chain, and the second TCR subunit is a TCR γ chain, wherein the first connecting peptide comprises the amino acid sequence of SEQ ID NO: 11, and the second connecting peptide comprises the amino acid sequence of SEQ ID NO: 12; or
(ii) the first TCR subunit is a TCR γ chain, and the second TCR subunit is a TCR δ chain, wherein the first connecting peptide comprises the amino acid sequence of SEQ ID NO: 12, and the second connecting peptide comprises the amino acid sequence of SEQ ID NO: 11.