THERAPEUTIC AND DIAGNOSTIC CLONED MHC-UNRESTRICTED RECEPTOR SPECIFIC FOR THE MUC1 TUMOR ASSOCIATED ANTIGEN
The invention provides an isolated nucleic acid encoding a receptor, other than an immunoglobulin, wherein the receptor binds to a MUC1 tumor antigen independently of an major histocompatibility complex (MHC). The invention provides a method of activating a signaling pathway and/or killing a cancer cell using a receptor that is similar to or is a T cell receptor
1 . An isolated nucleic acid encoding a receptor, other than an immunoglobulin, wherein the receptor binds to a MUC1 tumor antigen independently of a major histocompatibility complex (MHC).
2 . The isolated nucleic acid of claim 1 , wherein the receptor binds to MUC1 tumor antigen with about the same affinity for MUC1 as the MA TCR.
3 . The isolated nucleic acid of claim 1 , wherein the receptor binds to MUC1 tumor antigen with a higher affinity for MUC1 as the MA TCR.
4 . The isolated nucleic acid of claim 1 , wherein the receptor binds to MUC1 tumor antigen with a lower affinity for MUC1 as the MA TCR.
5 . The isolated nucleic acid of claim 1 , wherein the receptor is a T cell receptor.
6 - 12 . (canceled)
13 . The isolated nucleic acid of claim 1 , wherein the receptor is a scFv.
14 . The isolated nucleic acid of claim 1 , wherein the receptor is a T cell receptor (TCR).
15 . The isolated nucleic acid of claim 1 , wherein the receptor is a single chain receptor.
16 . The isolated nucleic acid of claim 1 , further comprising a linker of from 1 to about 30 amino acids between the first portion of the amino acid sequence and the second portion of the amino acid sequence.
17 . The isolated nucleic acid of claim 1 , wherein the receptor is expressed in a T cell better than an otherwise identical receptor lacking the linker in an identical T cell.
18 . The isolated nucleic acid of claim 1 , wherein the linker improves the expression of the receptor in a cell in comparison to an otherwise identical receptor lacking the linker.
19 . The isolated nucleic acid of claim 1 , wherein the linker does not have a function selected from the group consisting of an immunological function, a membrane spanning function, a signaling function, and a dimerization function.
20 . The isolated nucleic acid of claim 1 , wherein the Kd of the TCR for a single MUC1 epitope is between 0.2 μM and 200 μM.
21 . The isolated nucleic acid of claim 1 , wherein the receptor is soluble.
22 . The isolated nucleic acid of claim 1 . wherein the receptor is membrane bound.
23 . The isolated nucleic acid of claim 1 , wherein a cell transduced with the isolated nucleic acid and expressing the isolate nucleic acid has an avidity (k d ) for a cancer cell expressing the MUC1 tumor antigen of from about 1×10 −5 M to about 1×10 −12 M.
24 . The isolated nucleic acid of claim 1 , wherein the encoded receptor does not comprise a constant domain of an antibody.
25 . A cell transduced with the nucleic acid of claim 1 .
26 . A lymphocyte transduced with the nucleic acid of claim 1 .
27 . The cell of claim 25 , wherein the cell is a T cell.
28 . The cell of claim 25 , wherein the cell is a B cell.
29 . The cell of claim 25 , wherein the cell is selected from the group consisting of an NK cell, a macrophage, and a dendritic cell.
30 . The cell of claim 25 , wherein the cell is granulocyte.
31 . A cell other than a T cell or B cell comprising a receptor encoded by the nucleic acid of claim 1 .
32 . A population of cells isolated from an animal wherein the cells comprise a receptor encoded by the nucleic acid of claim 1 .
33 . A composition comprising:
the nucleic acid of claim 1 , and
a sterile carrier, pharmaceutically acceptable excipient, adjuvant, and/or buffer that is substantially isotonic.
34 . The composition claim 33 , wherein the composition is suitable for administration to a mammal.
35 . The composition of claim 34 , wherein the mammal is human.
36 . A composition comprising:
the cell of claim 25 , and
a sterile carrier, pharmaceutically acceptable excipient, adjuvant, and/or buffer that is substantially isotonic.
37 . The composition claim 36 , wherein the composition is suitable for administration to a mammal.
38 . The composition of claim 38 , wherein the mammal is human.
39 . A composition comprising:
the population of cells of claim 31 , and
a sterile carrier, pharmaceutically acceptable excipient, adjuvant, and/or buffer that is substantially isotonic.
40 . The composition claim 39 , wherein the composition is suitable for administration to a mammal.
41 . The composition of claim 40 , wherein the mammal is human.
42 . A gene delivery vector comprising the nucleic acid of claim 1 .
43 . The gene delivery vector of claim 42 , wherein the gene delivery vector is a viral vector.
44 . The gene delivery vector of claim 42 , wherein the viral vector is a retroviral vector.
45 . The gene delivery vector of claim 42 , wherein the gene delivery vector is selected from the group consisting of a herpes viral vector, an adenoviral vector, and an adeno-associated viral vector.
46 . The gene delivery vector of claim 42 , wherein the gene delivery vector is a lentiviral vector.
47 . The gene delivery vector of claim 42 , wherein the gene delivery vector is an “MFG” vector.
48 . The gene delivery vector of claim 42 , wherein the gene delivery vector is a non-viral vector.
49 . The gene delivery vector of claim 42 , wherein the gene delivery vector is a liposomal vector.
50 . An isolated or substantially purified receptor encoded by the nucleic acid of claim 1 .
51 . The isolated or substantially purified receptor of claim 50 , wherein the receptor is soluble.
52 . The isolated or substantially purified receptor of claim 50 , wherein the receptor is membrane bound.
53 . An immunocytochemistry stain comprising the receptor of claim 50 complexed with a labeling agent.
54 . An immunocytochemistry stain comprising the receptor of claim 51 complexed with a labeling agent.
55 . An immunocytochemistry stain comprising the receptor of claim 52 complexed with a labeling agent.
56 . (canceled)
57 . A method of activating a signaling pathway in a cell having a signaling pathway, the method comprising:
a. transducing the cell having a signaling pathway with at least one nucleic acid encoding a receptor, wherein the receptor
i. is expressed and displayed on the surface of the transduced cell,
ii. binds to a MUC1 tumor antigen independently of an major histocompatibility complex (MHC), and
b. contacting the transduced cell to a cell expressing the MUC1 tumor antigen thereby activating the signaling pathway.
58 . The method of claim 57 , wherein the receptor is a T cell receptor.
59 . A method of activating a signaling pathway in a cell having a signaling pathway, the method comprising:
a. transducing the cell having a signaling pathway with at least one nucleic acid encoding a receptor, which comprises the nucleic acid of claim 1 , wherein the receptor
i. is expressed and displayed on the surface of the transduced cell,
ii. binds to a MUC1 tumor antigen independently of an major histocompatibility complex (MHC), and
b. contacting the transduced cell to a cell expressing the MUC1 tumor antigen thereby activating the signaling pathway.
60 . The method of claim 59 , wherein the receptor is a T cell receptor.
61 . A method of activating a signaling pathway in a cell comprising a signaling pathway, the method comprising transducing the cell with a receptor having affinity for MUC1, wherein the affinity is determined by a first amino acid sequence and a second amino acid sequence, wherein
the first amino acid sequence consists essentially of the portion of MA Vα23 shown in FIG. 1 (SEQ ID NO:1), and
the second amino acid sequence consists essentially of the portion of MA Vβ8.3 shown in FIG. 1 (SEQ ID NO:2).
62 . A method of killing a cancer cell, the method comprising
a. isolating a population of cells comprising a receptor, wherein the receptor binds to a MUC1 tumor antigen independently of an major histocompatibility complex (MHC), and
b. contacting the isolated population of cells to a cell expressing the MUC1 tumor antigen thereby killing the cancer cell.
63 . The method of claim 60 , wherein the population comprises T cells.
64 . The method of claim 61 , wherein the population consists essentially of T cells.
65 . The method of claim 62 , wherein the population does not comprise cells, other than T cells, that comprise a receptor that binds to a MUC1 tumor antigen independently of a major histocompatibility complex (MHC).