Spycatcher and spytag: universal immune receptors for t cells

Abstract

The invention provides compositions and methods for adoptive T cell therapy in treating a variety of disorders including cancer, infections, and autoimmune disorders. In one embodiment, the invention provides a universal immune receptor that comprises a protein or peptide tag, such as a SpyCatcher or a SpyTag moiety, bound to an extracellular hinge region, a transmembrane domain, and an intracellular domain for T cell activation.

What is claimed: 
     
         1 . An isolated nucleic acid sequence encoding a universal immune receptor, wherein the universal immune receptor comprises either a SpyCatcher or a SpyTag extracellular binding domain bound to an extracellular hinge region, which is in turn bound to a transmembrane domain which is in turn bound to a T cell receptor intracellular signaling domain. 
     
     
         2 . The isolated nucleic acid sequence of  claim 1 , wherein a SpyCatcher extracellular binding domain is bound to the extracellular hinge domain. 
     
     
         3 . The isolated nucleic acid sequence of  claim 1 , wherein a SpyTag extracellular binding domain is bound to the extracellular hinge domain. 
     
     
         4 . The isolated nucleic acid sequence of  claim 1 , wherein the universal immune receptor comprises a nucleic acid sequence selected from the group consisting of SEQ ID Nos: 1, 3, 5, 7, 9, 11, 13 and 15. 
     
     
         5 . The isolated nucleic acid sequence of  claim 2 , wherein the universal immune receptor comprises a nucleic acid sequence selected from the group consisting of SEQ ID Nos: 5, 7, 13 and 15. 
     
     
         6 . The isolated nucleic acid sequence of  claim 3 , wherein the universal immune receptor comprises a nucleic acid sequence selected from the group consisting of SEQ ID Nos: 1, 3, 9 and 11. 
     
     
         7 . The isolated nucleic acid sequence of any one of  claims 1 - 3 , wherein the T cell receptor intracellular signaling domain further comprises a costimulatory molecule. 
     
     
         8 . The isolated nucleic acid sequence of any one of  claims 1 - 3 , wherein the T cell receptor intracellular signaling domain substantially lacks signaling capacity. 
     
     
         9 . The isolated nucleic acid sequence of  claim 8 , wherein the intracellular domain of the costimulatory molecule is selected from the group consisting of CD27, CD28, CD2, CD3, 4-1BB, OX40, CD30, CD40, PD-1, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD7, LIGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83, and any combination thereof. 
     
     
         10 . An isolated nucleic acid sequence encoding either a SpyCatcher or a SpyTag linked to a molecule comprising at least one selected from the group consisting of an oligonucleotide, an antibody, an antibody fragment, a scFv, a protein scaffold, a peptide, a ligand, an aptamer, a labelling agent, a tumor antigen, a self-antigen, a viral antigen, and any combination thereof. 
     
     
         11 . The isolated nucleic acid sequence of  claim 10 , wherein the labelling agent is selected from the group consisting of myc-tag, FLAG-tag, His-tag, HA-tag, a fluorescent protein (e.g. green fluorescent protein (GFP)), a fluorophore (e.g. tetramethylrhodamine (TRITC), fluorescein isothiocyanate (FITC)), dinitrophenol, peridinin chlorophyll protein complex, green fluorescent protein, phycoerythrin (PE), histidine, biotin, streptavidin, avidin, horse radish peroxidase, palmitoylation, nitrosylation, alkalanine phosphatase, glucose oxidase, Glutathione S-transferase (GST), maltose binding protein, a radioisotope, and any types of compounds used for radioisotope labeling including, 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA), diethylene triamine pentaacetic acid (DTPA), and 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA). 
     
     
         12 . An isolated universal immune receptor comprising either a SpyCatcher or a SpyTag extracellular binding domain bound to an extracellular hinge region, which is in turn bound to a transmembrane domain which is in turn bound to a T cell receptor intracellular signaling domain. 
     
     
         13 . The isolated universal immune receptor of  claim 12 , wherein a SpyCatcher extracellular binding domain is bound to the extracellular hinge domain. 
     
     
         14 . The isolated universal immune receptor of  claim 12 , wherein a SpyTag extracellular binding domain is bound to the extracellular hinge domain. 
     
     
         15 . The isolated universal immune receptor of  claim 12 , wherein the universal immune receptor comprises an amino acid sequence selected from the group consisting of SEQ ID Nos: 2, 4, 6, 8, 10, 12, 14 and 16. 
     
     
         16 . The isolated universal immune receptor of  claim 13 , wherein the universal immune receptor comprises an amino acid sequence selected from the group consisting of SEQ ID Nos: 6, 8, 14 and 16. 
     
     
         17 . The isolated universal immune receptor of  claim 14 , wherein the universal immune receptor comprises an amino acid sequence selected from the group consisting of SEQ ID Nos: 2, 4, 10 and 12. 
     
     
         18 . The isolated universal immune receptor of any one of  claims 12 - 14 , wherein the T cell receptor intracellular signaling domain further comprises a costimulatory molecule. 
     
     
         19 . The isolated universal immune receptor of  claim 18 , wherein the intracellular domain of the costimulatory molecule is selected from the group consisting of CD27, CD28, CD2, CD3, 4-1BB, OX40, CD30, CD40, PD-1, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD7, LIGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83, and any combination thereof. 
     
     
         20 . The isolated universal immune receptor of any one of  claims 12 - 14 , wherein the isolated universal immune receptor is bound to a composition comprising either a SpyTag- or a SpyCatcher-bound molecule. 
     
     
         21 . The isolated universal immune receptor of  claim 20 , wherein the SpyTag- or SpyCatcher-bound molecule comprises at least one selected from the group consisting of an oligonucleotide, an antibody, an antibody fragment, a scFv, a protein scaffold, a peptide, a ligand, an aptamer, a labelling agent, a tumor antigen, a self-antigen, a viral antigen, and any combination thereof. 
     
     
         22 . The isolated universal immune receptor of  claim 21 , wherein the labelling agent is selected from the group consisting of myc-tag, FLAG-tag, His-tag, HA-tag, a fluorescent protein (e.g. green fluorescent protein (GFP)), a fluorophore (e.g. tetramethylrhodamine (TRITC), fluorescein isothiocyanate (FITC)), dinitrophenol, peridinin chlorophyll protein complex, phycoerythrin (PE), histidine, biotin, streptavidin, avidin, horse radish peroxidase, palmitoylation, nitrosylation, alkalanine phosphatase, glucose oxidase, Glutathione S-transferase (GST), maltose binding protein, a radioisotope, and any types of compounds used for radioisotope labeling including, 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA), diethylene triamine pentaacetic acid (DTPA), and 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA). 
     
     
         23 . A cell comprising a nucleic acid sequence of sequence encoding a universal immune receptor, wherein the universal immune receptor comprises either a SpyCatcher or a SpyTag extracellular binding domain bound to an extracellular hinge region, which is in turn bound to a transmembrane domain which is in turn bound to a T cell receptor intracellular signaling domain. 
     
     
         24 . The cell of  claim 23 , wherein a SpyCatcher extracellular binding domain is bound to the extracellular hinge domain. 
     
     
         25 . The cell of  claim 23 , wherein a SpyTag extracellular binding domain is bound to the extracellular hinge domain. 
     
     
         26 . The cell of any one of  claims 23 - 25 , wherein the cell is selected from the group consisting of a T cell, a Natural Killer (NK) cell, a cytotoxic T lymphocyte (CTL), a macrophage, a stem cell, and a regulatory T cell. 
     
     
         27 . The cell of any one of  claims 23 - 25 , wherein the cell is activated when the encoded isolated universal immune receptor binds to a composition comprising either a SpyTag- or a SpyCatcher-bound molecule. 
     
     
         28 . The cell of  claim 27 , wherein the SpyTag- or SpyCatcher-bound molecule comprises at least one selected from the group consisting of an oligonucleotide, an antibody, an antibody fragment, a scFv, a protein scaffold, a peptide, a receptor, a ligand, an aptamer, a labelling agent, a tumor antigen, a self-antigen, a viral antigen, and any combination thereof. 
     
     
         29 . The cell of  claim 28 , wherein the labelling agent is selected from the group consisting of myc-tag, FLAG-tag, His-tag, HA-tag, a fluorescent protein (e.g. green fluorescent protein (GFP)), a fluorophore (e.g. tetramethylrhodamine (TRITC), fluorescein isothiocyanate (FITC)), dinitrophenol, peridinin chlorophyll protein complex, phycoerythrin (PE), histidine, biotin, streptavidin, avidin, horse radish peroxidase, palmitoylation, nitrosylation, alkalanine phosphatase, glucose oxidase, Glutathione S-transferase (GST), maltose binding protein, a radioisotope, and any types of compounds used for radioisotope labeling including, 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA), diethylene triamine pentaacetic acid (DTPA), and 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA). 
     
     
         30 . A vector comprising a nucleic acid sequence encoding a universal immune receptor, wherein the universal immune receptor comprises either a SpyCatcher or a SpyTag extracellular binding domain bound to an extracellular hinge region, which is in turn bound to a transmembrane domain which is in turn bound to a T cell receptor intracellular signaling domain. 
     
     
         31 . The vector of  claim 30 , wherein a SpyCatcher extracellular binding domain is bound to the extracellular hinge domain. 
     
     
         32 . The vector of  claim 30 , wherein a SpyTag extracellular binding domain is bound to the extracellular hinge domain. 
     
     
         33 . The vector of any one of  claims 30 - 32 , wherein the encoded isolated universal immune receptor can bind to a composition comprising either a SpyTag- or a SpyCatcher-bound molecule. 
     
     
         34 . The vector of  claim 33 , wherein the SpyTag- or SpyCatcher-bound molecule comprises at least one selected from the group consisting of an oligonucleotide, an antibody, an antibody fragment, a scFv, a protein scaffold, a peptide, a receptor, a ligand, an aptamer, a labelling agent, a tumor antigen, a self-antigen, a viral antigen, and any combination thereof. 
     
     
         35 . The vector of  claim 34 , wherein the labelling agent is selected from the group consisting of myc-tag, FLAG-tag, His-tag, HA-tag, fluorescent protein (e.g. green fluorescent protein (GFP)), a fluorophore (e.g. tetramethylrhodamine (TRITC), fluorescein isothiocyanate (FITC)), dinitrophenol, peridinin chlorophyll protein complex, green fluorescent protein, phycoerythrin (PE), histidine, biotin, streptavidin, avidin, horse radish peroxidase, palmitoylation, nitrosylation, alkalanine phosphatase, glucose oxidase, Glutathione S-transferase (GST), maltose binding protein, a radioisotope, and any types of compounds used for radioisotope labeling including, 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA), diethylene triamine pentaacetic acid (DTPA), and 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA). 
     
     
         36 . A method for stimulating a universal immune receptor-mediated immune response in a mammal, the method comprising administering to a mammal an effective amount of a cell genetically modified to express a universal immune receptor, wherein the universal immune receptor comprises either a SpyCatcher or a SpyTag extracellular binding domain bound to an extracellular hinge region, which is in turn bound to a transmembrane domain which is in turn bound to a T cell receptor intracellular signaling domain. 
     
     
         37 . The method of  claim 36 , wherein a SpyCatcher extracellular binding domain is bound to the extracellular hinge domain. 
     
     
         38 . The method of  claim 36 , wherein a SpyTag extracellular binding domain is bound to the extracellular hinge domain. 
     
     
         39 . The method of  claim 36 , wherein the universal immune receptor comprises an amino acid sequence selected from the group consisting of SEQ ID Nos: 2, 4, 6, 8, 10, 12, 14 and 16. 
     
     
         40 . The method of  claim 37 , wherein the universal immune receptor comprises an amino acid sequence selected from the group consisting of SEQ ID Nos: 6, 8, 14 and 16. 
     
     
         41 . The method of  claim 38 , wherein the universal immune receptor comprises an amino acid sequence selected from the group consisting of SEQ ID Nos: 2, 4, 10 and 12. 
     
     
         42 . The method of claim any one of  claims 36 - 38 , wherein the universal immune receptor further comprises an intracellular domain of a costimulatory molecule. 
     
     
         43 . The method of  claim 42 , wherein the intracellular domain of a costimulatory molecule is selected from the group consisting of CD27, CD28, CD2, CD3, 4-1BB, OX40, CD30, CD40, PD-1, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD7, LIGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83, and any combination thereof. 
     
     
         44 . The method of any one of  claims 36 - 38 , wherein the universal immune receptor-mediated immune response is stimulated when the isolated universal immune receptor binds to a composition comprising either a SpyTag- or a SpyCatcher-bound molecule. 
     
     
         45 . The method of  claim 44 , wherein the SpyTag- or SpyCatcher-bound molecule comprises at least one selected from the group consisting of an oligonucleotide, an antibody, an antibody fragment, a scFv, a protein scaffold, a peptide, a receptor, a ligand, an aptamer, a labelling agent, a tumor antigen, a self-antigen, a viral antigen, and any combination thereof. 
     
     
         46 . The method of  claim 45 , wherein the labelling agent is selected from the group consisting of myc-tag, FLAG-tag, His-tag, HA-tag, a fluorescent protein (e.g. green fluorescent protein (GFP)), a fluorophore (e.g. tetramethylrhodamine (TRITC), fluorescein isothiocyanate (FITC)), dinitrophenol, peridinin chlorophyll protein complex, green fluorescent protein, phycoerythrin (PE), histidine, biotin, streptavidin, avidin, horse radish peroxidase, palmitoylation, nitrosylation, alkalanine phosphatase, glucose oxidase, Glutathione S-transferase (GST), maltose binding protein, a radioisotope, and any types of compounds used for radioisotope labeling including, 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA), diethylene triamine pentaacetic acid (DTPA), and 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA). 
     
     
         47 . The method of any one of  claims 36 - 38 , wherein the method comprises administering either a SpyTag- or a SpyCatcher-bound molecule to the mammal prior to administering the genetically modified cell to the mammal. 
     
     
         48 . The method of any one of  claims 36 - 38 , wherein the method comprises administering a plurality of SpyTag- or SpyCatcher-bound molecules to the mammal prior to administering the genetically modified cell to the mammal. 
     
     
         49 . The method of any one of  claims 36 - 38 , wherein the method comprises sequentially administering a plurality of SpyTag- or SpyCatcher-bound molecules to the mammal prior to administering the genetically modified cell to the mammal. 
     
     
         50 . The method of any one of  claims 36 - 38 , wherein the method comprises binding the universal immune receptor with either a SpyTag- or a SpyCatcher-bound molecule prior to administering the genetically modified cell to the mammal. 
     
     
         51 . The method of any one of  claims 36 - 38 , wherein the method comprises binding the universal immune receptor with a plurality of SpyTag- or SpyCatcher-bound molecules prior to administering the genetically modified cell to the mammal. 
     
     
         52 . The method of any one of  claims 36 - 38 , wherein the method comprises sequentially binding the universal immune receptor with a plurality of SpyTag- or SpyCatcher-bound molecules prior to administering the genetically modified cell to the mammal. 
     
     
         53 . The method of any one of  claims 36 - 38 , wherein the cell is an autologous cell. 
     
     
         54 . The method of any one of  claims 36 - 38 , wherein the cell is selected from the group consisting of a T cell, a Natural Killer (NK) cell, a cytotoxic T lymphocyte (CTL), a macrophage, a stem cell, and a regulatory T cell. 
     
     
         55 . A method of treating a mammal in need thereof, the method comprising administering to the mammal an effective amount of the cell of  claim 23 . 
     
     
         56 . The method of any one of  claim 36  or  55 , wherein the mammal is treated for a disorder selected from the group consisting of a viral, a bacterial and a parasitic infection, an autoimmune disease and a cancer.