Constructs targeting AFP peptide/MHC complexes and uses thereof
The present application provides constructs comprising an antibody moiety that specifically binds to a complex comprising an AFP peptide and an MHC class I protein. Also provided are methods of making and using these constructs.
1. An isolated anti-AMC construct comprising an antibody moiety that specifically binds to a complex comprising an alpha-fetoprotein (AFP) peptide and a major histocompatibility (MHC) class I protein (AFP/MHC class I complex, or AMC), wherein the AFP peptide comprises the amino acid sequence of SEQ ID NO: 4, wherein the MHC class I protein is HLA-A02, and wherein the antibody moiety comprises:
a heavy chain variable domain comprising a heavy chain complementarity determining region (HC-CDR)1 comprising the amino acid sequence of SEQ ID NO: 63, an HC-CDR2 comprising the amino acid sequence of SEQ ID NO: 73, and an HC-CDR3 comprising the amino acid sequence of SEQ ID NO: 83; and
a light chain variable domain comprising a light chain complementarity determining region (LC-CDR)1 comprising the amino acid sequence of SEQ ID NO: 96, an LC-CDR2 comprising the amino acid sequence of SEQ ID NO: 106, and an LC-CDR3 comprising the amino acid sequence of SEQ ID NO: 116.
2. The isolated anti-AMC construct of claim 1 , wherein the heavy chain variable domain comprises the amino acid sequence of SEQ ID NO: 23 and the light chain variable domain comprises the amino acid sequence of SEQ ID NO: 33.
3. The isolated anti-AMC construct of claim 1 , wherein the isolated anti-AMC construct is a chimeric antigen receptor (CAR) comprising an extracellular domain comprising the antibody moiety, a transmembrane domain, and an intracellular signaling domain capable of activating an immune cell.
4. The isolated anti-AMC construct of claim 3 , wherein the intracellular signaling domain comprises a CD3ζ intracellular signaling sequence and a co-stimulatory signaling sequence.
5. The isolated anti-AMC construct of claim 4 , wherein the co-stimulatory signaling sequence comprises a CD28 or 4-1BB intracellular signaling sequence.
6. The isolated anti-AMC construct of claim 5 , wherein the co-stimulatory signaling sequence comprises a CD28 intracellular signaling sequence.
7. The isolated anti-AMC construct of claim 6 , wherein the transmembrane domain comprises a CD28 transmembrane region.
8. The isolated anti-AMC construct of claim 3 , wherein the antibody moiety is an scFv.
9. The isolated anti-AMC construct of claim 1 , wherein the isolated anti-AMC construct is a tandem scFv comprising a first scFv linked by a peptide linker to a second scFv, wherein the first scFv is the antibody moiety that specifically binds to the AFP/MHC class I complex.
10. The isolated anti-AMC construct of claim 9 , wherein the second scFv is specific for CD3ε.
11. The isolated anti-AMC construct of claim 10 , wherein the first scFv is N-terminal to the second scFv.
12. The isolated anti-AMC construct of claim 1 , wherein the isolated anti-AMC construct is an immunoconjugate comprising the antibody moiety and an effector molecule, wherein the effector molecule is a therapeutic agent selected from the group consisting of a drug, a toxin, a radioisotope, a protein, a peptide, and a nucleic acid.
13. The isolated anti-AMC construct of claim 1 , wherein the isolated anti-AMC construct is an immunoconjugate comprising the antibody moiety and a label.
14. A nucleic acid encoding the polypeptide components of the isolated anti-AMC construct of claim 1 .
15. A host cell expressing the isolated anti-AMC construct of claim 1 .
16. An effector cell expressing the isolated anti-AMC construct of claim 1 .
17. A CAR T cell expressing the isolated anti-AMC construct of claim 3 , wherein the intracellular signaling domain is capable of activating a T cell.
18. A pharmaceutical composition comprising the CART cell of claim 17 .
19. A method of treating an individual having an AFP-positive cancer, comprising administering to the individual an effective amount of the pharmaceutical composition of claim 18 , wherein the cancer expresses human AFP and HLA-A02.
20. The method of claim 19 , wherein the administration is via intravenous or intratumoral route.
21. The method of claim 19 , wherein the cancer is hepatocellular carcinoma, germ cell tumor, or breast cancer.
22. The method of claim 21 , wherein the cancer is hepatocellular carcinoma.
23. The method of claim 22 , wherein the cancer is metastatic hepatocellular carcinoma.
24. The isolated anti-AMC construct of claim 1 , wherein the MHC class I protein is the HLA-A*02:01 subtype of the HLA-A02 allele.