Non-hla restricted t cell receptors and uses thereof

Abstract

The presently disclosed subject matter provides methods and compositions for enhancing the immune response toward cancers and pathogens. It relates to novel designs of T cell receptors (TCRs) and engineered immunoresponsive cells comprising the same. The novel TCR binds to an antigen in an HLA-independent manner. In certain embodiments, the novel TCR provides enhanced sensitivity for a target gene having a low expression level.

What is claimed is: 
     
         1 . A recombinant T cell receptor (TCR) comprising an antigen binding chain that comprises an extracellular antigen-binding domain and a constant domain, wherein the recombinant TCR binds to an antigen in an HLA-independent manner, wherein the constant domain comprises a native or modified TRAC peptide and/or a native or modified TRBC peptide. 
     
     
         2 . The recombinant TCR of  claim 1 , wherein the recombinant TCR is expressed from an expression cassette placed in an endogenous TRAC locus and/or a TRBC locus of an immunoresponsive cell. 
     
     
         3 . The recombinant TCR of  claim 2 , wherein the placement of the expression cassette (a) disrupts or abolishes the endogenous expression of a TCR comprising a native TCR α chain and/or a native TCR β chain in the immunoresponsive cell; or (b) prevents or eliminates mispairing between the recombinant TCR and a native TCR α chain and/or a native TCR β chain in the immunoresponsive cell. 
     
     
         4 . The recombinant TCR of  claim 1 , wherein the constant domain is capable of forming a homodimer or a heterodimer with another constant domain. 
     
     
         5 . The recombinant TCR of  claim 1 , wherein the antigen binding chain is capable of associating with a CD3ζ polypeptide. 
     
     
         6 . The recombinant TCR of  claim 5 , wherein the antigen binding chain, upon binding to an antigen, is capable of activating the CD3ζ polypeptide associated to the antigen binding chain. 
     
     
         7 . The recombinant TCR of  claim 6 , wherein activation of the CD3ζ polypeptide is capable of activating an immunoresponsive cell. 
     
     
         8 . The recombinant TCR of  claim 5 , wherein the CD3ζ polypeptide is endogenous and integrated in the native CD3 complex. 
     
     
         9 . The recombinant TCR of  claim 5 , wherein the CD3ζ polypeptide is exogenous and optionally integrated with a co-stimulatory molecule selected from the group consisting of a CD28 polypeptide, a 4-1BB polypeptide, an OX40 polypeptide, an ICOS polypeptide, a DAP-10 polypeptide and any combination thereof. 
     
     
         10 . The recombinant TCR of  claim 1 , wherein the antigen binding chain further comprises a co-stimulatory region, wherein the recombinant TCR, upon binding to an antigen, is capable of stimulating an immunoresponsive cell. 
     
     
         11 . The recombinant TCR of  claim 10 , wherein the co-stimulatory region comprises a co-stimulatory molecule selected from the group consisting of a CD28 polypeptide, a 4-1BB polypeptide, an OX40 polypeptide, an ICOS polypeptide, a DAP-10 polypeptide and any combination thereof. 
     
     
         12 . The recombinant TCR of  claim 11 , wherein the co-stimulatory region comprises a CD28 polypeptide. 
     
     
         13 . The recombinant TCR of  claim 1 , wherein the recombinant TCR is capable of integrating with a CD3ζ complex and providing HLA-independent antigen recognition. 
     
     
         14 . The recombinant TCR of  claim 13 , wherein the CD3ζ complex is endogenous. 
     
     
         15 . The recombinant TCR of  claim 14 , wherein the recombinant TCR replaces an endogenous TCR in a CD3/TCR complex. 
     
     
         16 . The recombinant TCR of  claim 1 , wherein the extracellular antigen-binding domain is capable of dimerizing with another extracellular antigen-binding domain. 
     
     
         17 . The recombinant TCR of  claim 1 , wherein the extracellular antigen-binding domain comprises a ligand for a cell-surface receptor, a receptor for a cell surface ligand, an antigen binding portion of an antibody, or an antigen binding portion of a TCR. 
     
     
         18 . The recombinant TCR of  claim 1 , wherein the extracellular antigen-binding domain comprises one or two immunoglobulin variable region(s). 
     
     
         19 . The recombinant TCR of  claim 17 , wherein the extracellular antigen-binding domain comprises a heavy chain variable region (V H ) of an antibody, and/or a light chain variable region (V L ) of an antibody. 
     
     
         20 . The recombinant TCR of  claim 1 , wherein the extracellular antigen-binding domain is capable of dimerizing with another extracellular antigen-binding domain. 
     
     
         21 . The recombinant TCR of  claim 20 , wherein the extracellular antigen-binding domain comprises (a) a V H  of an antibody, wherein the V H  is capable of dimerizing with another extracellular antigen-binding domain comprising a V L  of the antibody and form a fragment variable (Fv); and/or a V L  of an antibody, wherein the V L  is capable of dimerizing with another extracellular antigen-binding domain comprising a V H  of the antibody and form a fragment variable (Fv). 
     
     
         22 . The recombinant TCR of  claim 1 , wherein the recombinant TCR binds to a tumor antigen. 
     
     
         23 . The recombinant TCR of  claim 22 , wherein the tumor antigen is selected from the group consisting of CD19, MUC16, MUC1, CA1X, CEA, CD8, CD7, CD10, CD20, CD22, CD30, CLL1, CD33, CD34, CD38, CD41, CD44, CD49f, CD56, CD74, CD133, CD138, EGP-2, EGP-40, EpCAM, erb-B2,3,4, FBP, Fetal acetylcholine receptor, folate receptor-a, GD2, GD3, HER-2, hTERT, IL-13R-a2, K-light chain, KDR, LeY, L1 cell adhesion molecule, MAGE-A1, Mesothelin, ERBB2, MAGEA3, p53, MART 1, GP100, Proteinase3 (PR1), Tyrosinase, Survivin, hTERT, EphA2, NKG2D ligands, NY-ES0-1, oncofetal antigen (h5T4), PSCA, PSMA, ROR1, TAG-72, VEGF-R2, WT-1, BCMA, CD123, CD44V6, NKCS1, EGF1R, EGFR-VIII, and CD99, CD70, ADGRE2, CCR1, LILRB2, LILRB4, PRAME, and ERBB. 
     
     
         24 . The recombinant TCR of  claim 23 , wherein the tumor antigen is CD19. 
     
     
         25 . The recombinant TCR of  claim 1 , wherein the recombinant TCR exhibits greater antigen sensitivity than a CAR targeting the same antigen, and/or is capable of inducing an immune response when binds to an antigen that has a low density on the surface of a tumor cell. 
     
     
         26 . The recombinant TCR of  claim 1 , wherein the antigen that has a low density on the cell surface has a density of less than about 10,000 molecules per cell. 
     
     
         27 . An immunoresponsive cell comprising a recombinant TCR that comprises an antigen binding chain that comprises an extracellular antigen-binding domain and a constant domain, wherein the recombinant TCR binds to an antigen in an HLA-independent manner, wherein the constant domain comprises a native or modified TRAC peptide and/or a native or modified TRBC peptide. 
     
     
         28 . The immunoresponsive cell of  claim 27 , wherein at least one antigen binding chain of the recombinant TCR is expressed from an expression cassette placed at an endogenous gene locus of the immunoresponsive cell, optionally wherein two antigen binding chains of the recombinant TCR are expressed from expression cassettes placed at the endogenous gene locus of the immunoresponsive cell, wherein the two antigen binding chains are capable of dimerization. 
     
     
         29 . The immunoresponsive cell of  claim 28 , wherein the endogenous gene locus is a CD3δ locus, a CD3ε locus, a CD247 locus, a B2M locus, a TRAC locus, a TRBC locus, a TRDC locus and/or a TRGC locus. 
     
     
         30 . The immunoresponsive cell of  claim 29 , wherein the endogenous gene locus is a TRAC locus and/or a TRBC locus. 
     
     
         31 . The immunoresponsive cell of  claim 28 , wherein the placement of the expression cassette of the recombinant TCR disrupts or abolishes the endogenous expression of a TCR comprising a native TCR α chain and/or a native TCR β chain in the immunoresponsive cell, whereby preventing or eliminating mispairing between the recombinant TCR and a native TCR α chain and/or a native TCR β chain in the immunoresponsive cell. 
     
     
         32 . The immunoresponsive cell of  claim 28 , wherein the endogenous gene locus comprises a modified transcription terminator region. 
     
     
         33 . The immunoresponsive cell of  claim 32 , wherein the modified transcription terminator region comprises a genomic element selected from the group consisting of a TK transcription terminator, a GCSF transcription terminator, a TCRA transcription terminator, an HBB transcription terminator, a bovine growth hormone transcription terminator, an SV40 transcription terminator, and a P2A element. 
     
     
         34 . The immunoresponsive cell of  claim 28 , wherein when one endogenous T cell receptor locus in a cell is modified to express the at least one antigen binding chain of the recombinant TCR, at least one additional endogenous T cell receptor locus in the cell is modified to eliminate the expression of an endogenous TCR chain. 
     
     
         35 . The immunoresponsive cell of  claim 34 , wherein the at least one additional endogenous T cell receptor locus is further modified to express a gene of interest, optionally wherein the gene of interest is an anti-tumor cytokine, a co-stimulatory molecule ligand, a tracking gene or a suicide gene. 
     
     
         36 . The immunoresponsive cell of  claim 27 , wherein the immunoresponsive cell is selected from the group consisting of a T cell, a human embryonic stem cell, and a pluripotent stem cell from which lymphoid cells may be differentiated. 
     
     
         37 . The immunoresponsive cell of  claim 36 , wherein the immunoresponsive cell is a T cell. 
     
     
         38 . The immunoresponsive cell of  claim 37 , wherein the T cell is selected from the group consisting of a cytotoxic T lymphocyte (CTL), a regulatory T cell, and a Natural Killer T (NKT) cell. 
     
     
         39 . The immunoresponsive cell of  claim 27 , further comprising at least one exogenous co-stimulatory ligand. 
     
     
         40 . The immunoresponsive cell of  claim 39 , wherein the exogenous co-stimulatory ligand is selected from the group consisting of CD80, CD86, 41BBL, CD275, CD40L, OX40L, and combinations thereof. 
     
     
         41 . The immunoresponsive cell of  claim 39 , wherein (a) the cell further comprises or consists of one exogenous co-stimulatory ligand, optionally wherein the one exogenous co-stimulatory ligand is CD80 or 4-1BBL; or (b) the cell further comprises or consists of two exogenous co-stimulatory ligands, optionally wherein the two exogenous co-stimulatory ligands are CD80 and 4-1BBL. 
     
     
         42 . The immunoresponsive cell of  claim 27 , further comprising at least one chimeric costimulatory receptor (CCR). 
     
     
         43 . The immunoresponsive cell of  claim 42 , wherein the CCR comprises a co-stimulatory molecule selected from the group consisting of a CD28 polypeptide, a 4-1BB polypeptide, an OX40 polypeptide, an ICOS polypeptide, a DAP-10 polypeptide, and combinations thereof. 
     
     
         44 . A pharmaceutical composition comprising an effective amount of an immunoresponsive cell of  claim 27  and a pharmaceutically acceptable excipient, optionally wherein the pharmaceutical composition is for treating a neoplasm. 
     
     
         45 . A method of reducing tumor burden in a subject, and/or treating or preventing a neoplasm in a subject, and/or lengthening survival of a subject having a neoplasm, the method comprising administering to the subject an effective amount of the immunoresponsive cells of  claim 27 . 
     
     
         46 . A method for producing an antigen-specific immunoresponsive cell, the method comprising introducing into an immunoresponsive cell a nucleic acid molecule encoding a recombinant TCR comprising an antigen binding chain that comprises an extracellular antigen-binding domain and a constant domain, wherein the recombinant TCR binds to an antigen in an HLA-independent manner, wherein the constant domain comprises a native or modified TRAC polypeptide and/or a native or modified TRBC polypeptide. 
     
     
         47 . The method of  claim 46 , wherein at least one antigen binding chain of the recombinant TCR is expressed from an expression cassette placed at an endogenous gene locus of the immunoresponsive cell, optionally wherein two antigen binding chains of the recombinant TCR are expressed from expression cassettes placed at the endogenous gene locus of the immunoresponsive cell, wherein the two antigen binding chains are capable of dimerization. 
     
     
         48 . The method of  claim 47 , wherein the endogenous gene locus is a CD3δ locus, a CD3ε locus, a CD247locus, a B2M locus, a TRAC locus, a TRBC locus, a TRDC locus and/or a TRGC locus, optionally wherein wherein the endogenous gene locus is a TRAC locus and/or a TRBC locus. 
     
     
         49 . The method of  claim 47 , wherein the endogenous gene locus comprises a modified transcription terminator region, optionally wherein the modified transcription terminator region comprises a genomic element selected from the group consisting of a TK transcription terminator, a GCSF transcription terminator, a TCRA transcription terminator, a HBB transcription terminator, a bovine growth hormone transcription terminator, an SV40 transcription terminator, and a P2A element. 
     
     
         50 . The method of  claim 47 , wherein when one endogenous T cell receptor locus in a cell is modified to express the at least one antigen binding chain of the recombinant TCR, at least one additional endogenous T cell receptor locus in the cell is modified to eliminate the expression of an endogenous TCR chain, optionally wherein the at least one additional endogenous T cell receptor locus is further modified to express a gene of interest, optionally wherein the gene of interest is an anti-tumor cytokine, a co-stimulatory molecule ligand, a tracking gene or a suicide gene. 
     
     
         51 . A nucleotide acid molecule encoding a recombinant TCR of  claim 1 . 
     
     
         52 . A vector comprising the nucleic acid molecule of  claim 51 . 
     
     
         53 . A kit comprising a recombinant TCR of  claim 1 , optionally wherein the kit further comprises written instructions for treating and/or preventing a neoplasm, a pathogen infection, an autoimmune disorder, or an allogeneic transplant.