Therapeutic and diagnostic cloned mhc-unrestricted receptor specific for the muc1 tumor associated antigen

Abstract

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).