Chimeric antigen receptors recognizing cancer-specific TN glycopeptide variants
Disclosed are binding proteins, or fragments thereof, that specifically binds to a cancer-specific glycosylation variant of a protein and to a second epitope on the same protein, to a different protein presented on the same cell, or to a different protein presented on a different cell, such as an encoded polypeptide binding to both a cancer cell and an activated T cell. Also disclosed are polynucleotides encoding such binding proteins, including polynucleotides comprising codon-optimized coding regions and polynucleotides comprising coding regions that are not codon-optimized for expression in a particular host cell. Also disclosed are methods of making the encoded polypeptide and methods of using the polypeptide to treat, prevent or ameliorate the symptom of a disease such as cancer.
1. A cancer-specific Tn glycopeptide binding partner that binds a cancer-specific Tn glycopeptide, wherein the binding partner comprises the antibody heavy chain variable fragment (VH) of SEQ ID NO:19 or a humanized derivative thereof, and the antibody light chain variable fragment (VL) of SEQ ID NO:20 or a humanized derivative thereof.
2. The cancer-specific Tn glycopeptide binding partner of claim 1 wherein the binding partner comprises the antibody heavy chain variable fragment (VH) of SEQ ID NO:19 and the antibody light chain variable fragment (VL) of SEQ ID NO:20.
3. The cancer-specific Tn glycopeptide binding partner of claim 1 wherein the binding partner is a single-chain variable fragment (scFv).
4. The cancer-specific Tn glycopeptide binding partner of claim 3 wherein the scFv comprises the heavy chain variable fragment N-terminal to the light chain variable fragment.
5. The cancer-specific Tn glycopeptide binding partner of claim 3 wherein the scFv heavy chain variable fragment and light chain variable fragment are covalently bound to a linker sequence of 4-15 amino acids.
6. The cancer-specific Tn glycopeptide binding partner of claim 3 wherein the single-chain variable fragment is contained within a bi-specific T-cell engager.
7. The cancer-specific Tn glycopeptide binding partner of claim 3 wherein the single-chain variable fragment is contained within a chimeric antigen receptor.
8. A polynucleotide comprising a coding region for a cancer-specific Tn glycopeptide binding partner heavy chain variable region and light chain variable region according to claim 1 .
9. The polynucleotide according to claim 8 , wherein the coding region is codon-optimized for expression in a human cell.
10. The polynucleotide according to claim 9 wherein the coding region for the heavy chain variable fragment is set forth in SEQ ID NO:9 and the coding region for the light chain variable fragment is set forth in SEQ ID NO:11.
11. The polynucleotide according to claim 8 wherein the polynucleotide encodes a cancer-specific Tn glycopeptide binding partner selected from the group consisting of a single-chain variable fragment, a multimer of a single-chain variable fragment, a bi-specific single-chain variable fragment and a multimer of a bi-specific single-chain variable fragment.
12. The polynucleotide according to claim 11 wherein the multimer of a single-chain variable fragment is selected from the group consisting of a divalent single-chain variable fragment, a tribody and a tetrabody.
13. The polynucleotide according to claim 11 wherein the multimer of a bi-specific single-chain variable fragment is a bi-specific T-cell engager.
14. The polynucleotide according to claim 8 further comprising a coding region for a peptide selected from the group consisting of a peptide signaling domain of a T cell signaling protein, a peptide modulator of T cell activation, and an enzymatic component of a labeling system.
15. The polynucleotide according to claim 14 wherein the peptide signaling domain of a T cell signaling protein is selected from the group consisting of a 4-1 BB cytosolic signaling domain, a CD3 cytosolic signaling domain, a cytosolic domain of CD28-CD3 fusion and a cytosolic domain of a 4-1 BB-CD3ζ fusion.
16. The polynucleotide according to claim 8 further comprising a peptide modulator of T cell activation selected from the group consisting of IL15, IL15Rα and an IL15/IL15Rα fusion peptide.
17. The polynucleotide according to claim 8 further comprising a coding region for a linker peptide comprising the sequence of SEQ ID NO:14.
18. The polynucleotide according to claim 8 further comprising a coding region for a signal peptide comprising the sequence of SEQ ID NO:8.