Anti-CD276 polypeptides, proteins, and chimeric antigen receptors
Polypeptides and proteins that specifically bind to and immunologically recognize CD276 are disclosed. Chimeric antigen receptors (CARs), anti-CD276 binding moieties, nucleic acids, recombinant expression vectors, host cells, populations of cells, and pharmaceutical compositions relating to the polypeptides and proteins are also disclosed. Methods of detecting the presence of cancer in a mammal and methods of treating or preventing cancer in a mammal are also disclosed.
1. A polypeptide comprising a heavy and light chain comprising (i) SEQ ID NOs: 1-6, a heavy and light chain comprising (ii) SEQ ID NOs: 11-16, or a heavy and light chain comprising (iii) SEQ ID NOs: 20-25, wherein the heavy and light chains of (i), (ii), or (iii) bind to CD276.
2. The polypeptide of claim 1 comprising (i) SEQ ID NOs: 7 and 8, (ii) SEQ ID NOs: 17 and 18, or (iii) SEQ ID NOs: 26 and 27.
3. A protein comprising a first polypeptide chain comprising (i) a heavy chain comprising SEQ ID NOs: 1-3, (ii) a heavy chain comprising SEQ ID NOs: 11-13, or (iii) a heavy chain comprising SEQ ID NOs: 20-22 and a second polypeptide chain comprising (iv) a light chain comprising SEQ ID NOs: 4-6, (v) a light chain comprising SEQ ID NOs: 14-16, or (vi) a light chain comprising SEQ ID NOs: 23-25, wherein the first polypeptide of (i), (ii), or (iii) and the second polypeptide of (iv), (v), or (vi) bind to CD276.
4. The protein of claim 3 comprising a first polypeptide chain comprising SEQ ID NO: 7, 17, or 26 and a second polypeptide chain comprising SEQ ID NO: 8, 18, or 27.
5. The polypeptide of claim 1 further comprising a linker amino acid sequence.
6. The polypeptide of claim 5 , wherein the linker comprises SEQ ID NO: 115.
7. The polypeptide of claim 1 comprising SEQ ID NO: 10, 19, or 28.
8. A chimeric antigen receptor (CAR) comprising
(a) an antigen binding domain formed by the polypeptide of claim 1 ,
(b) a transmembrane domain, and
(c) an intracellular T cell signaling domain.
9. The CAR according to claim 8 , wherein the transmembrane domain comprises CD8 and/or CD28.
10. The CAR according to claim 8 , wherein the transmembrane domain comprises any one or more of a CD8 amino acid sequence comprising SEQ ID NO: 29, a CD28 amino acid sequence comprising SEQ ID NO: 30, and a CD8 amino acid sequence comprising SEQ ID NO: 31.
11. The CAR according to claim 8 , wherein the intracellular T cell signaling domain comprises one or more of i) CD28, ii) CD137, and iii) CD3 zeta.
12. The CAR according to claim 8 , wherein the intracellular T cell signaling domain comprises a CD28 amino acid sequence comprising SEQ ID NO: 32 and/or SEQ ID NO: 35.
13. The CAR according to claim 8 , wherein the intracellular T cell signaling domain comprises a CD137 amino acid sequence comprising SEQ ID NO: 33 and/or SEQ ID NO: 37.
14. The CAR according to claim 8 , wherein the intracellular T cell signaling domain comprises a CD3 zeta amino acid sequence comprising any one or more of SEQ ID NOs: 34, 36, and 38.
15. The CAR according to claim 8 comprising an amino acid sequence comprising any one of SEQ ID NOs: 39-47 and 122-130.
16. An anti-CD276 binding moiety comprising the polypeptide of claim 1 , wherein the anti-CD276 binding moiety is an antibody, Fab fragment (Fab), F(ab′) 2 fragment, diabody, triabody, tetrabody, single-chain variable region fragment (scFv), or disulfide-stabilized variable region fragment (dsFv).
17. A conjugate comprising (a) the polypeptide of claim 1 conjugated to (b) an effector molecule.
18. The conjugate of claim 17 , wherein the effector molecule is a drug, toxin, label, small molecule, or an antibody.
19. The conjugate of claim 17 , wherein the effector molecule is PE4E, PE40, PE38, PE25, PE38QQR, PE38KDEL, PE-LR, or PE35.
20. A nucleic acid comprising a nucleotide sequence encoding the polypeptide of claim 1 .
21. The nucleic acid according to claim 20 , comprising a nucleotide sequence selected from the group consisting of SEQ ID NOs: 57-59.
22. The nucleic acid according to claim 21 , comprising a nucleotide sequence encoding a CAR and selected from the group consisting of SEQ ID NOs: 48-56 and 131-139.
23. The nucleic acid according to claim 20 , comprising a nucleotide sequence comprising (i) SEQ ID NOs: 116 and 117, (ii) SEQ ID NOs: 118 and 119, or (iii) SEQ ID NOs: 120 and 121.
24. A recombinant expression vector comprising the nucleic acid of claim 20 .
25. An isolated host cell comprising the recombinant expression vector of claim 24 .
26. A population of cells comprising at least one isolated host cell of claim 25 .
27. A pharmaceutical composition comprising the polypeptide of claim 1 and a pharmaceutically acceptable carrier.
28. A method of detecting the presence of cancer in a mammal, comprising:
(a) contacting a sample comprising one or more cells from the mammal with the polypeptide of claim 1 thereby forming a complex, and
(b) detecting the complex, wherein detection of the complex is indicative of the presence of cancer in the mammal.
29. The polypeptide of claim 1 expressed in one or more host cells for use in treating cancer in a mammal.