Dimeric antigen receptors
The present disclosure provides dimeric antigen receptors (DAR) constructs comprising a heavy chain binding region on one polypeptide chain and a light chain binding region on a separate polypeptide chain. The two polypeptide chains that make up the dimeric antigen receptors can dimerize to form an antigen binding domain. The dimeric antigen receptors have antibody-like properties as they bind specifically to a target antigen. The dimeric antigen receptors can be used for directed cell therapy.
1. A nucleic acid encoding a precursor polypeptide comprising ten regions ordered from the amino terminus to the carboxyl terminus: (1) a heavy chain leader sequence, (2) an antibody heavy chain variable region comprising a CD38 antibody heavy chain variable region which comprises the amino acid sequence of SEQ ID NO:1, (3) an antibody heavy chain constant region, (4) an optional hinge region, (5) a transmembrane region, (6) an intracellular signaling region, (7) a T2A cleavage sequence, (8) a light chain leader sequence, (9) an antibody light chain variable region comprising a CD38 antibody light chain variable region which comprises the amino acid sequence of SEQ ID NO:3, and (10) an antibody light chain constant region.
2. The nucleic acid of claim 1 , comprising (4) the hinge region.
3. The nucleic acid of claim 1 , wherein (1) the heavy chain leader sequence comprises the amino acid sequence of SEQ ID NO: 10; (3) the antibody heavy chain constant region comprises a CD38 antibody heavy chain constant region which comprises the amino acid sequence of SEQ ID NO:2; (4) the hinge region is selected from a group consisting of a CD28 hinge region comprising the amino acid sequence of SEQ ID NO:5, a CD8 hinge region comprising the amino acid sequence of SEQ ID NO:21, and a hinge region comprising a CD28 hinge region comprising the amino acid sequence of SEQ ID NO:5 and a CD8 hinge region comprising the amino acid sequence of SEQ ID NO:21; (5) the transmembrane region comprises a CD28 transmembrane region which comprises the amino acid sequence of SEQ ID NO:6; (6) the intracellular signaling region comprises any one or any combination of two or more signaling sequences selected from a group consisting of 4-1BB signaling sequence comprising the amino acid sequence of SEQ ID NO:7, CD28 signaling sequence comprising the amino acid sequence of SEQ ID NO:8, CD3zeta (long) signaling sequence comprising the amino acid sequence of SEQ ID NO:9 and/or CD3zeta (short) signaling sequence having an ITAM 3 motif and comprising the amino acid sequence of SEQ ID NO:20; (7) the T2A cleavage sequence comprises the amino acid sequence of SEQ ID NO:12; (8) the light chain leader sequence comprises the amino acid sequence of SEQ ID NO: 11; and (10) the antibody light chain constant region comprises a CD38 antibody light chain constant region which comprises the amino acid sequence of SEQ ID NO:4.
4. A vector operably linked to the nucleic acid of claim 1 .
5. A host cell or a population of host cells harboring the vector of claim 4 .
6. The host cell of claim 5 , wherein the vector is an expression vector, and wherein the expression vector directs transcription and/or translation (expression) of the nucleic acid.
7. The host cell or the population of host cells of claim 5 , wherein the host cell or the population of host cells comprise T lymphocytes (T cells, regulatory T cells, gamma-delta T cells or cytotoxic T cells), NK (natural killer) cells, macrophages, dendritic cells, mast cells, eosinophils, B lymphocytes or monocytes.
8. A method for treating a subject having a disease, disorder or condition associated with detrimental expression of a tumor antigen in the subject, comprising: administering to the subject the host cell or the population of host cells of claim 5 .
9. The method of claim 8 , wherein the disease is a hematologic cancer selected from the group consisting of non-Hodgkin's lymphoma (NHL), Burkitt's lymphoma (BL), B chronic lymphocytic leukemia (B-CLL), B and T acute lymphocytic leukemia (ALL), T cell lymphoma (TCL), acute myeloid leukemia (AML), hairy cell leukemia (HCL), Hodgkin's Lymphoma (HL), chronic myeloid leukemia (CML) and multiple myeloma (MM).
10. A method for treating a subject having a disease, disorder or condition associated with detrimental expression of a tumor antigen in the subject, comprising: administering to the subject the host cell or the population of host cells of claim 6 .