Affinity-enhanced monomeric streptavidin chimeric antigen receptor (CAR)
A chimeric antigen receptor is disclosed that includes: (a) an extracellular high affinity streptavidin;(b) a hinge domain from CD8; (c) a CD28 transmembrane domain; (d) an intracellular 4-1BB and/or CD28 signaling domain; and (e) an intracellular CD3 zeta signaling domain, wherein (a)-(e) are in N-terminal to C-terminal order. Nucleic acids encoding this chimeric antigen receptor, and T and natural killer (NK) cells transformed with this chimeric antigen receptor are also disclosed. The use of this chimeric antigen receptor for the treatment of tumors is also disclosed.
1. A method for treating a subject with a tumor, comprising
administering to the subject a therapeutically effective amount of a monoclonal antibody that specifically binds a tumor associated antigen expressed by the tumor, wherein the monoclonal antibody is biotinylated, and
administering to the subject a pharmaceutical composition, thereby treating the tumor in the subject,
wherein the pharmaceutical composition comprises i) an effective amount of an expression vector comprising a promoter operably linked to a nucleic acid molecule encoding a chimeric antigen receptor comprising amino acids 1-369 of SEQ ID NO: 11 or amino acids 1-371 of SEQ ID NO: 12 or ii) an effective amount of CD3+T cells and/or natural killer cells transduced with the expression vector of (i), and a pharmaceutically acceptable carrier, thereby treating the tumor in the subject.
2. The method of claim 1 , wherein the pharmaceutical composition comprises the CD3+T cells, and wherein the T cells are autologous to the subject.
3. The method of claim 1 , wherein
a) the pharmaceutical composition comprises the CD3+ T cells, wherein the CD3+ T cells are CD3 + CD4 + T cells and/or CD3 + CD8 + T cells; and/or
b) the tumor associated antigen is MUC-1, CD19 or CD20.
4. The method of claim 1 , wherein the subject is human.
5. The method of claim 1 , wherein a) the tumor is a lymphoma or leukemia; or b) the tumor is a solid tumor.
6. The method of claim 1 , wherein the chimeric antigen receptor comprises amino acids 1-371 of SEQ ID NO: 12.
7. The method of claim 1 , wherein the expression vector comprises the nucleic acid sequence of SEQ ID NO: 13.
8. The method of claim 1 , wherein the expression vector comprises the nucleic acid sequence of SEQ ID NO: 14.
9. The method of claim 1 , wherein the expression vector comprises (a) the nucleic acid sequence of SEQ ID NO: 15; and/or (b) the nucleic acid of SEQ ID NO: 16.
10. The method of claim 1 , wherein the expression vector comprises the nucleic acid sequence of SEQ ID NO: 17 or SEQ ID NO: 18.
11. The method of claim 1 , wherein the expression vector comprises the nucleic acid sequence of SEQ ID NO: 19.
12. The method of claim 1 , wherein the expression vector is codon-optimized for expression in human cells.
13. The method of claim 1 , wherein the expression vector comprises the nucleic acid sequence of SEQ ID NO: 20 or SEQ ID NO: 21.
14. The method of claim 1 , wherein the expression vector is a viral vector.
15. The method of claim 14 , wherein the viral vector is a lentiviral vector or a gamma retroviral vector.
16. The method of claim 1 , wherein the tumor associated antigen is EGFR or HER2.
17. A method for treating a subject with a tumor, comprising
transducing CD3+ T cells and/or natural killer cells from the subject with an expression vector comprising a promoter operably linked to a nucleic acid molecule encoding a chimeric antigen receptor comprising amino acids 1-369 of SEQ ID NO: 11 or amino acids 1-371 of SEQ ID NO: 12 to produce autologous transduced cells that express the chimeric antigen receptor:
administering to the subject a therapeutically effective amount of a monoclonal antibody that specifically binds a tumor associated antigen expressed by the tumor, wherein the monoclonal antibody is biotinylated; and
administering to the subject a therapeutically effective amount of the autologous transduced cells that express the chimeric antigen receptor, thereby treating the tumor in the subject.
18. The method of claim 17 , wherein the tumor associated antigen is MUC-1, CD19 or CD20.
19. The method of claim 17 , wherein the expression vector is a gamma retroviral vector.
20. The method of claim 17 , wherein the tumor associated antigen is EGFR or HER2.