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 V H and C H 1 antibody domains 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 V L and C L antibody domains and a second TCRD comprising a second transmembrane domain of a second TCR subunit,
wherein the V H and C H 1 domains of the first antigen-binding domain and the V L and C L domains of the second antigen-binding domain form an antigen-binding module that specifically binds to CD19,
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,
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,
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,
and wherein 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.
2. The abTCR 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 abTCR 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 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.
5. 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.
6. 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.
7. The abTCR of claim 5 , 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.
8. The abTCR of claim 7 , 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.
9. The abTCR of claim 7 , 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.
10. A composition comprising nucleic acid(s) encoding the first and second polypeptide chains of the abTCR of claim 1 .
11. 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.
12. An effector cell presenting on its surface the abTCR of claim 1 .
13. An effector cell comprising the nucleic acid(s) of the composition of claim 10 .
14. The effector cell of claim 12 , wherein the effector cell does not express TCRγ and/or TCRδ.
15. The effector cell of claim 14 , wherein the effector cell is an αβ T cell.
16. The effector cell of claim 12 , wherein the effector cell is modified to block or decrease the expression of a first endogenous TCR subunit and/or a second endogenous TCR subunit.
17. The effector cell of claim 12 , wherein the effector cell is selected from the group consisting of a cytotoxic T cell, a helper T cell, a natural killer T cell, and a suppressor T cell.
18. The effector cell of claim 12 , comprising a vector comprising a first nucleic acid sequence encoding the first polypeptide chain of the abTCR and a second nucleic acid sequence encoding the second polypeptide chain of the abTCR.
19. A pharmaceutical composition comprising the effector cell of claim 12 .
20. 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 19 .
21. The method of claim 20 , wherein the CD19-associated disease is cancer.
22. The method of claim 21 , wherein the cancer is lymphoma, or leukemia.
23. The abTCR of claim 1 , wherein the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 56, and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 54.
24. An antibody-T cell receptor (TCR) chimeric molecule (abTCR) that specifically binds to a complex comprising an AFP peptide and a major histocompatibility complex (MHC) protein, comprising:
a) a first polypeptide chain comprising a first antigen-binding domain comprising Vu and C H 1 antibody domains 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 V L and C L antibody domains and a second TCRD comprising a second transmembrane domain of a second TCR subunit,
wherein the V H and C H 1 domains of the first antigen-binding domain and the V L and C L domains of the second antigen-binding domain form an antigen-binding module that specifically binds to the complex comprising the AFP peptide and the MHC protein,
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,
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, 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,
and wherein the V H antibody domain comprises the amino acid sequence of SEQ ID NO: 38 and the V L antibody domain comprises the amino acid sequence of SEQ ID NO: 40.
25. The abTCR of claim 24 , wherein the first TCR subunit is a TCR γ chain, and the second TCR subunit is a TCR δ chain.
26. The abTCR of claim 24 , wherein the first TCR subunit is a TCR δ chain, and the second TCR subunit is a TCR γ chain.
27. The abTCR of claim 24 , 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.
28. The abTCR of claim 24 , 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.
29. The abTCR of claim 24 , 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.
30. The abTCR of claim 28 , 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.
31. The abTCR of claim 30 , 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.
32. The abTCR of claim 30 , 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.
33. The abTCR of claim 24 , wherein the first polypeptide chain comprises the amino acid sequence of SEQ ID NO: 35, and the second polypeptide chain comprises the amino acid sequence of SEQ ID NO: 36.
34. A composition comprising nucleic acid(s) encoding the first and second polypeptide chains of the abTCR of claim 24 .
35. A complex comprising the abTCR of claim 24 and at least one TCR-associated signaling module selected from the group consisting of CD3δε, CD3γε, and ζζ.
36. An effector cell presenting on its surface the abTCR of claim 24 .
37. An effector cell comprising the nucleic acid(s) of the composition of claim 34 .
38. The effector cell of claim 36 , wherein the effector cell does not express TCRγ and/or TCRδ.
39. The effector cell of claim 38 , wherein the effector cell is an αβ T cell.
40. The effector cell of claim 36 , wherein the effector cell is modified to block or decrease the expression of a first endogenous TCR subunit and/or a second endogenous TCR subunit.
41. The effector cell of claim 36 , wherein the effector cell is selected from the group consisting of a cytotoxic T cell, a helper T cell, a natural killer T cell, and a suppressor T cell.
42. The effector cell of claim 36 , comprising a vector comprising a first nucleic acid sequence encoding the first polypeptide chain of the abTCR and a second nucleic acid sequence encoding the second polypeptide chain of the abTCR.
43. A pharmaceutical composition comprising the effector cell of claim 36 .
44. A method of treating an AFP-associated disease in an individual in need thereof comprising administering to the individual an effective amount of the pharmaceutical composition of claim 43 .
45. The method of claim 44 , wherein the AFP-associated disease is cancer.
46. The method of claim 45 , wherein the cancer is hepatocellular carcinoma.