Chimeric antigen receptor compositions
Provided herein are compositions, which exhibit 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) a first portion comprising an antibody heavy chain variable domain and an antibody heavy chain constant domain;
(ii) a second portion comprising an antibody light chain variable domain and an antibody light chain constant domain;
(iii) a self-cleaving peptidyl sequence between said first portion and said second portion; and
(iv) a CD3 ξ intracellular T-cell signaling domain;
wherein said first portion further comprises a transmembrane domain; and
wherein the nucleic acid sequence encoding said second portion is 3′ to the nucleic acid sequence encoding said first portion.
2. The isolated nucleic acid of claim 1 , wherein said protein comprises from the amino terminus to the carboxy terminus: said heavy chain variable domain, said heavy chain constant domain, said transmembrane domain and said second portion.
3. The isolated nucleic acid of claim 2 , wherein said antibody heavy chain variable domain and said antibody light chain variable domain are humanized.
4. The isolated nucleic acid of claim 1 , wherein said first portion further comprises an intracellular co-stimulatory signaling domain.
5. The isolated nucleic acid of claim 4 , 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.
6. The isolated nucleic acid of claim 4 , further comprising a spacer region positioned between said heavy chain variable domain and said transmembrane domain.
7. The isolated nucleic acid of claim 6 , wherein said spacer region further comprises a hinge region.
8. The isolated nucleic acid of claim 6 , wherein said first portion further comprises a linker domain.
9. The isolated nucleic acid of claim 8 , wherein said linker domain is between said transmembrane domain and said intracellular T-cell signaling domain.
10. The isolated nucleic acid of claim 9 , wherein said linker domain is between said intracellular T-cell signaling domain and said intracellular co-stimulatory signaling domain.
11. The isolated nucleic acid of claim 8 , wherein said linker domain comprises the sequence GGCGG (SEQ ID NO: 121) or GGG.
12. The isolated nucleic acid of claim 1 , wherein said self-cleaving peptidyl encoding sequence is a T2A encoding sequence or a 2A encoding sequence.
13. The isolated nucleic acid of claim 1 , wherein said protein is an anti-CD19 protein, anti-CD20 protein, anti-CD22 protein, anti-CD30 protein, anti-CD33 protein, anti-CD44v6/7/8 protein, anti-CD123 protein, anti-CEA protein, anti-EGP-2 protein, anti-EGP-40 protein, anti-erb-B2 protein, anti-erb-B3 protein, anti-erb-B4 protein, anti-FBP protein, anti-fetal acetylcholine receptor protein, anti-GD2 protein, anti-GD3 protein, anti-Her2/neu protein, anti-IL-13R-a2 protein, anti-KDR protein, anti k-light chain protein, anti-LeY protein, anti-L1 cell adhesion molecule protein, anti-MAGE-A1 protein, anti-mesothelin protein, anti-murine CMV infected cell protein, anti-MUC2 protein, anti-NKGD2 protein, anti-oncofetal antigen protein, anti-PCSA protein, anti-PSMA protein, anti-TAA protein, anti-EGFR protein, anti-TAG-72 protein or anti-VEGF-72 protein.
14. An expression vector comprising said nucleic acid of claim 1 .
15. A T lymphocyte comprising said expression vector of claim 14 .
16. A T lymphocyte comprising said protein of claim 1 .
17. A method of treating cancer, said method comprising administering to a subject in need thereof an effective amount of said T-lymphocyte of claim 16 , wherein said antibody heavy chain variable domain and said antibody light chain variable domain form part of an anti-cancer antibody region.