Chimeric antigen receptor compositions
Provided herein are compositions which exhibit novel diagnostic capabilities and allow to rapidly add functionality to adoptive immunotherapy. The compositions include isolated nucleic acids encoding proteins including antibody regions capable of binding compounds including a peptidyl moiety (e.g., a meditope). The recombinant proteins provided herein are useful, inter alia, for a broad variety of therapeutic and diagnostic purposes. For example, the recombinant proteins provided herein including embodiments thereof may be used as non-invasive means to characterize chimeric antigen receptor (CAR) T cells before and/or during treatment of diseases (e.g., cancer).
1. An isolated nucleic acid encoding a protein comprising:
(i) an antibody region comprising a central cavity formed by a heavy chain variable (VH) region, a light chain variable (VL) region, a heavy chain constant region (CH) and a light chain constant region (CL), wherein said central cavity forms a peptide binding site comprising framework region amino acid residues; and
(ii) a transmembrane domain.
2. The isolated nucleic acid of claim 1 , wherein said antibody region is an antibody fragment.
3. The isolated nucleic acid of claim 1 , wherein said antibody region comprises an Fc domain.
4. The isolated nucleic acid of claim 1 , wherein said antibody region is a humanized antibody region.
5. The isolated nucleic acid of claim 1 , further comprising a nucleic acid sequence encoding an intracellular T-cell signaling domain.
6. The isolated nucleic acid of claim 5 , wherein said intracellular T-cell signaling domain is a CD3 ζ intracellular T-cell signaling domain.
7. The isolated nucleic acid of claim 6 , further comprising a nucleic acid sequence encoding an intracellular co-stimulatory signaling domain.
8. The isolated nucleic acid of claim 7 , wherein said intracellular co-stimulatory signaling domain is a CD28 intracellular co-stimulatory signaling domain, a 4-1BB intracellular co-stimulatory signaling domain, a ICOS intracellular co-stimulatory signaling domain, or an OX-40 intracellular co-stimulatory signaling domain.
9. The isolated nucleic acid of claim 8 , further comprising a spacer sequence encoding a spacer region.
10. The isolated nucleic acid of claim 9 , wherein said spacer region is between said transmembrane domain and said antibody region.
11. The isolated nucleic acid of claim 10 , further comprising a linker sequence encoding a linker domain.
12. The isolated nucleic acid of claim 11 , wherein said linker domain is between said transmembrane domain and said intracellular T-cell signaling domain.
13. The isolated nucleic acid of claim 11 , wherein said linker domain is between said intracellular T-cell signaling domain and said intracellular co-stimulatory signaling domain.
14. The isolated nucleic acid of claim 11 , wherein said linker domain comprises the sequence GGCGG or GGG.
15. The isolated nucleic acid of claim 1 comprising:
(i) a heavy chain sequence encoding a heavy chain domain of said protein, said heavy chain domain comprising a variable heavy chain domain and said transmembrane domain; and
(ii) a light chain sequence encoding a light chain domain of said protein, said light chain domain comprising a variable light chain domain, wherein said variable heavy chain domain and said variable light chain domain together form at least a portion of said antibody region.
16. The isolated nucleic acid of claim 15 encoding a self-cleaving peptidyl sequence between said heavy chain sequence and said light chain sequence.
17. The isolated nucleic acid of claim 16 , wherein said self-cleaving peptidyl linker sequence is a T2A sequence or a 2A sequence.
18. The isolated nucleic acid of 17 , wherein said light chain sequence is 3′ to said heavy chain sequence.
19. The isolated nucleic acid of 17 , wherein said light chain sequence is 5′ to said heavy chain sequence.
20. A T lymphocyte comprising the isolated nucleic acid of claim 1 .