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 effector cell comprising one or more nucleic acid sequence(s) encoding 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, and a first TCR 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, 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 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; and
wherein the first TCRD and the second TCRD form a TCR module (TCRM) that is capable of recruiting at least one TCR-associated signaling module.
2. The effector cell of claim 1 , wherein the effector cell presents the abTCR on its surface.
3. The effector cell of claim 1 , 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.
4. The effector cell 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 effector cell of claim 4 , wherein the first and/or second peptide linkers comprise, individually, a constant domain or fragment thereof from an immunoglobulin or a TCR subunit.
6. The effector cell of claim 5 , wherein the first and/or second peptide linkers comprise, individually, a C H 1, C H 2, C H 3, C H 4, or C L antibody domain, or a fragment thereof.
7. The effector cell of claim 5 , wherein the first and/or second peptide linkers comprise, individually, a Cα, Cβ, Cγ, or Cδ TCR domain, or a fragment thereof.
8. The effector cell of claim 1 , wherein the TCR-associated signaling module is selected from the group consisting of CD3δε, CD376, and ζζ.
9. The effector cell of claim 1 , wherein the first polypeptide chain lacks the variable domain of the first TCR subunit, and wherein the second polypeptide chain lacks the variable domain of the second TCR subunit.
10. The effector cell of claim 1 , wherein:
(i) the first TCR subunit is TCRγ, and the second TCR subunit is TCRδ; or
(ii) the first TCR subunit is TCRδ, and the second TCR subunit is TCRγ; and
wherein the effector cell is an αβ T cell, or a δγ T cell modified to block or decrease the expression of endogenous TCRγ and TCRδ.
11. The effector cell of claim 1 , wherein:
(i) the first TCR subunit is TCRα, and the second TCR subunit is TCRβ; or
(ii) the first TCR subunit is TCRβ, and the second TCR subunit is TCRα; and
wherein the effector cell is a δγ T cell, or a αβ T cell modified to block or decrease the expression of endogenous TCRα and TCRβ.
12. The effector cell of claim 1 , wherein:
(i) the first polypeptide chain comprises a C H 1 antibody domain, and the second polypeptide chain comprises a C L antibody domain; or
(ii) the second polypeptide chain comprises a C H 1 antibody domain, and the first polypeptide chain comprises a C L antibody domain.
13. The effector cell of claim 1 , wherein the first TCRD further comprises a first connecting peptide or fragment thereof of the first TCR subunit N-terminal to the first transmembrane domain, and the second TCRD further comprises a second connecting peptide or fragment thereof of the second TCR subunit N-terminal to the second transmembrane domain.
14. The effector cell of claim 13 , wherein the TCRM comprises a disulfide bond between a residue in the first connecting peptide and a residue in the second connecting peptide.
15. The effector cell of claim 13 , wherein:
(i) the first TCR subunit is TCRδ, and the second TCR subunit is TCRγ, 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;
(ii) the first TCR subunit is TCRγ, and the second TCR subunit is TCRδ, 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;
(iii) the first TCR subunit is TCRδ, and the second TCR subunit is TCRγ, 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
(iv) the first TCR subunit is TCRγ, and the second TCR subunit is TCRδ, 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.
16. The effector cell of claim 13 , wherein:
(i) the first TCR subunit is TCRα, and the second TCR subunit is TCRβ, wherein the first connecting peptide comprises the amino acid sequence of SEQ ID NO: 5, and the second connecting peptide comprises the amino acid sequence of SEQ ID NO: 6;
(ii) the first TCR subunit is TCRD, and the second TCR subunit is TCRα, wherein the first connecting peptide comprises the amino acid sequence of SEQ ID NO: 6, and the second connecting peptide comprises the amino acid sequence of SEQ ID NO: 5;
(iii) the first TCR subunit is TCRα, and the second TCR subunit is TCRβ, wherein the first connecting peptide comprises the amino acid sequence of SEQ ID NO: 9, and the second connecting peptide comprises the amino acid sequence of SEQ ID NO: 10; or
(iv) the first TCR subunit is TCRβ, and the second TCR subunit is TCRα, wherein the first connecting peptide comprises the amino acid sequence of SEQ ID NO: 10, and the second connecting peptide comprises the amino acid sequence of SEQ ID NO: 9.
17. A pharmaceutical composition comprising the effector cell of claim 1 , and a pharmaceutically acceptable carrier.
18. A method of treating a CD19-positive cancer in an individual in need thereof, comprising administering to the individual an effective amount of the pharmaceutical composition of claim 17 , wherein the effector cell specifically binds to CD19.
19. 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.
20. The abTCR of claim 10 , wherein the transmembrane domain of TCR comprises the amino acid sequence of SEQ ID NO: 3, and the transmembrane domain of TCRγ comprises the amino acid sequence of SEQ ID NO: 4.