Transfected T-cells and T-cell receptors for use in immunotherapy against cancers
Disclosed are T-cell receptors (TCRs) binding to tumor-associated antigens (TAAs) for targeting cancer cells, T-cells expressing same, methods for producing same, and methods for treating cancers using same. Disclosed are TCRs and their variants that bind to HLA class I or II molecules with a peptide, such as MAG-003 have the amino acid sequence of KVLEHVVRV (SEQ ID NO:1). The description further relates to peptides, proteins, nucleic acids, cells for use in immunotherapeutic methods, the immunotherapy of cancer, and tumor-associated T-cell peptide epitopes, alone or in combination with other tumor-associated peptides that can for example serve as active pharmaceutical ingredients of vaccine compositions that stimulate anti-tumor immune responses, or to stimulate T-cells ex vivo and transfer into patients. Peptides bound to molecules of the major histocompatibility complex (MHC), or peptides as such, can also be targets of antibodies, soluble T-cell receptors, and other binding molecules.
1. A single chain TCR comprising an alpha chain and a beta chain,
wherein the alpha chain comprises
SEQ ID NO: 42,
SEQ ID NO: 43,
SEQ ID NO: 44, and
the beta chain comprises
SEQ ID NO: 50,
SEQ ID NO: 51, and
SEQ ID NO: 52.
2. The single chain TCR according to claim 1 , wherein the single chain TCR specifically and/or selectively binds to an epitope consisting of the amino acid sequence SEQ ID NO: 1 in a complex with an MHC class I molecule.
3. The single chain TCR according to claim 1 , wherein said single chain TCR is capable of inducing an immune response in a human subject.
4. The single chain TCR according to claim 1 , wherein the CDR3 alpha chain comprises SEQ ID NO: 44 and wherein the CDR3 beta chain comprises SEQ ID NO: 52.
5. The single chain TCR according to claim 1 , wherein
the CDR1 alpha chain comprises SEQ ID NO: 42,
the CDR2 alpha chain comprises SEQ ID NO: 43,
the CDR3 alpha chain comprises SEQ ID NO: 44,
the CDR1 beta chain comprises SEQ ID NO: 50,
the CDR2 beta chain comprises SEQ ID NO: 51, and
the CDR3 beta chain comprises SEQ ID NO: 52.
6. The single chain TCR according to claim 1 , wherein
the CDR1 alpha chain consists of SEQ ID NO: 42,
the CDR2 alpha chain comprises SEQ ID NO: 43,
the CDR3 alpha chain consists of SEQ ID NO: 44,
the CDR1 beta chain consists of SEQ ID NO: 50,
the CDR2 beta chain comprises SEQ ID NO: 51, and
the CDR3 beta chain consists of SEQ ID NO: 52.
7. The single chain TCR according to claim 5 , comprising a TCR variable chain region having at least 95% sequence identity to SEQ ID NO: 41.
8. The single chain TCR according to claim 5 , wherein the construct is humanized, chimerized and/or murinized.
9. The single chain TCR according to claim 1 , wherein
the CDR1 alpha chain consists of SEQ ID NO: 42,
the CDR2 alpha chain consists of SEQ ID NO: 43,
the CDR3 alpha chain consists of SEQ ID NO: 44,
the CDR1 beta chain consists of SEQ ID NO: 50,
the CDR2 beta chain consists of SEQ ID NO: 51, and
the CDR3 beta chain consists of SEQ ID NO: 52.
10. The single chain TCR according to claim 7 , wherein said single chain TCR further comprises a TCR constant region having at least 95%, sequence identity to SEQ ID NO: 46.
11. The single chain TCR according to claim 1 , comprising an alpha TCR chain having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 39 and a beta TCR chain having at least sequence identity to the amino acid sequence of SEQ ID NO: 47.
12. The single chain TCR according to claim 5 , comprising an alpha TCR chain having 100% sequence identity to the amino acid sequence of SEQ ID NO: 39, and a TCR beta chain having 100% sequence identity to the amino acid sequence of SEQ ID NO: 47.
13. The single chain TCR according to claim 12 , wherein said single chain TCR specifically binds to a peptide consisting of the amino acid sequence of KVLEHVVRV (SEQ ID NO: 1) in a complex with an MHC class I molecule.
14. A nucleic acid encoding for an antigen recognizing construct according to claim 1 .
15. A vector comprising a nucleic acid according to claim 14 .
16. A host cell comprising an antigen recognizing construct according to claim 1 , or a nucleic acid encoding for said antigen recognizing construct, or a vector comprising said nucleic acid.
17. The host cell according to claim 16 , wherein the cell is a lymphocyte, optionally a T lymphocyte or T lymphocyte progenitor, optionally a CD4 or CD8 positive T-cell.
18. A pharmaceutical composition comprising the single chain TCR of claim 1 , and a pharmaceutical acceptable carrier, stabilizer and/or excipient.
19. The single chain TCR according to claim 5 , wherein the binding affinity (K D ) for the TCR binding to a peptide consisting of the amino acid sequence of KVLEHVVRV (SEQ ID NO: 1) in a complex with an MHC class I molecule is about 100 μM or less.
20. The single chain TCR according to claim 5 , wherein T-cells transformed with a nucleic acid encoding said single chain TCR exhibits interferon-gamma release after coincubation with a target cell loaded with a peptide consisting of the amino acid sequence of KVLEHVVRV (SEQ ID NO: 1), wherein the peptide has an EC50 value of about 10 nM.
21. The single chain TCR according to claim 12 , wherein T-cells transformed with a nucleic acid encoding said single chain TCR exhibits interferon-gamma release after coincubation with a target cell loaded with a peptide consisting of the amino acid sequence of KVLEHVVRV (SEQ ID NO: 1), wherein the peptide has an EC50 value of about 10 nM.