Methods and Compositions for Cells Expressing a Chimeric Intracellular Signaling Molecule

Abstract

The present invention relates to compositions and methods for enhancing T cell metabolism and activity for more effective adoptive T cell therapy. By expressing an intracellular signaling molecule in T cells, the T cells are metabolically enhanced with improved cytotoxicity and resistance to immunosuppression imposed by tumor microenvironments. One aspect includes a modified T cell and pharmaceutical compositions comprising the modified cells for adoptive cell therapy and treating a disease or condition associated with enhanced immunity.

1 . A metabolically enhanced T cell comprising a chimeric intracellular signaling molecule, wherein:
 (a) the chimeric intracellular signaling molecule comprises:
 (i) an intracellular domain of a co-stimulatory molecule, wherein the intracellular domain is selected from the intracellular domain of a T cell receptor (TCR), a CD3 zeta, a CD3 gamma, a CD3 delta, a CD3 epsilon, a CD86, a common FcRgamma, FcR beta, CD79a, CD79b, Fcgamma DAP10, DAP12, CD27, CD28, 4-1BB, OX40, CD30, CD40, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, NKG2D, B7-H3, a ligand that specifically binds with CD83, ICAM-1, GITR, BAFFR, LIGHTR, SLAMF7, NKp80, CD127, CD160, CD19, CD4, CD8alpha, CD8beta, IL2R beta, IL2R gamma, IL7R alpha, ITGA4, VLA1, CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, CD11a, CD11b, CD11c, CD11d, ITGAE, CD103, ITGAL, ITGAM, ITGAX, ITGB1, CD29, ITGB2, CD18, ITGB7, TNFR2, TRANCE/RANKL, CD226, CD244, CD84, CD96, CEACAM1, CRTAM, CD229, CD160, PSGL1, CD100, CD69, SLAMF6, SLAMF1, CD150, IPO-3, SLAMF8, CD162, LTBR, LAT, GADS, SLP-76, PAG/Cbp, NKp44, NKp30, or NKp46, and 
 (ii) a myristoylation signal, or 
 (iii) a hinge and a transmembrane domain, wherein the hinge and/or the transmembrane domain is selected from the hinge and/or transmembrane domain of an alpha, beta or zeta chain of a T-cell receptor, CD28, CD3 epsilon, CD45, CD4, CD5, CD8, CD8-alpha, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD154, KIR, OX40, CD2, CD27, LFA-1, ICOS, 4-1BB, GITR, CD40, BAFFR, HVEM, SLAMF7, NKp80, CD160, CD19, IL2R beta, IL2R gamma, IL7Ra, ITGA1, VLA1, CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, CD1 ld, ITGAE, CD103, ITGAL, CD1 1 a, ITGAM, CD1 lb, ITGAX, CD1 1 c, ITGB1, CD29, ITGB2, CD18, ITGB7, TNFR2, DNAM1, SLAMF4, CD84, CD96, CEACAM1, CRTAM, Ly9, CD160 (BY55), PSGL1, CD100, SLAMF1, SLAMF6, SLAMF8, SELPLG, LTBR, PAG, NKp44, NKp30, NKp46, NKG2D, NKG2C, or any combination thereof; 
   (b) the chimeric intracellular signaling molecule substantially lacks an extracellular ligand-binding domain; and   (c) the metabolically enhanced T cell expresses the chimeric intracellular signaling molecule.   
     
     
         2 . The metabolically enhanced T cell of  claim 1 , wherein expression of the chimeric intracellular signaling molecule metabolically enhances the T cell. 
     
     
         3 . The metabolically enhanced T cell of  claim 1 , wherein expression of the chimeric intracellular signaling molecule improves cytotoxicity and resistance to immunosuppression when in a tumor microenvironment. 
     
     
         4 . The metabolically enhanced T cell of  claim 1  further comprising a bispecific antibody. 
     
     
         5 . The metabolically enhanced T cell of  claim 4 , wherein the bispecific antibody comprises a first antigen binding domain that binds to a first antigen and a second antigen binding domain that binds to a second antigen. 
     
     
         6 . The metabolically enhanced T cell of  claim 5 , wherein the first antigen binding domain binds to a target cell and the second antigen binding domain binds to an activated T cell. 
     
     
         7 . The metabolically enhanced T cell of  claim 5 , wherein the bispecific antibody comprises an antigen binding domain comprising a first and a second single chain variable fragment (scFv) molecule. 
     
     
         8 . (canceled) 
     
     
         9 . A pharmaceutical composition comprising the metabolically enhanced T cell of  claim 1  and a pharmaceutically acceptable carrier. 
     
     
         10 - 30 . (canceled) 
     
     
         31 . A composition comprising the population of metabolically enhanced T cells of  claim 32 . 
     
     
         32 . A population of metabolically enhanced T cells comprising a chimeric intracellular signaling molecule, wherein:
 (a) the chimeric intracellular signaling molecule comprises:
 (i) an intracellular domain of a co-stimulatory molecule, wherein the intracellular domain is selected from the intracellular domain of a T cell receptor (TCR), CD3 zeta, CD3 gamma, a CD3 delta, CD3 epsilon, CD86, FcRgamma, FcR beta, CD79a, CD79b, Fcgamma RIIa, DAP10, DAP12, CD27, CD28, 4-1BB, OX40, CD30, CD40, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, NKG2D, B7-H3, a ligand that specifically binds with CD83, ICAM-1, GITR, BAFFR, LIGHTR, SLAMF7, NKp80, CD127, CD160, CD19, CD4, CD8alpha, CD8beta, IL2R beta, IL2R gamma, IL7R alpha, ITGA4, VLA1, CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, CD11a, CD11b, CD11 c, CD11d, ITGAE, CD103, ITGAL, ITGAM, ITGAX, ITGB1, CD29, ITGB2, CD18, ITGB7, TNFR2, TRANCE/RANKL, CD226, CD244, CD84, CD96, CEACAM1, CRTAM, CD229, CD160, PSGL1, CD100, CD69, SLAMF6, SLAMF1, CD150, IPO-3, SLAMF8, CD162, LTBR, LAT, GADS, SLP-76, PAG/Cbp, NKp44, NKp30, or NKp46, and 
   (ii) a myristoylation signal, or   (iii) a hinge and a transmembrane domain, wherein the hinge and/or the transmembrane domain is selected from the hinge and/or transmembrane domain of an alpha, beta or zeta chain of a T-cell receptor, CD28, CD3 epsilon, CD45, CD4, CD5, CD8, CD8-alpha, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD154, KIR, OX40, CD2, CD27, LFA-1, ICOS, 4-1BB, GITR, CD40, BAFFR, HVEM, SLAMF7, NKp80, CD160, CD19, IL2R beta, IL2R gamma, IL7Ra, ITGA1, VLA1, CD49a, ITGA4, IA4, CD49D, ITGA6, VLA-6, CD49f, ITGAD, CD11d, ITGAE, CD103, ITGAL, CD1 1 a, ITGAM, CD1 1 b, ITGAX, CD1 1 c, ITGB1, CD29, ITGB2, CD18, ITGB7, TNFR2, DNAM1, SLAMF4, CD84, CD96, CEACAM1, CRTAM, Ly9, CD160 (BY55), PSGL1, CD100, SLAMF1, SLAMF6, SLAMF8, SELPLG, LTBR, PAG, NKp44, NKp30, NKp46, NKG2D, NKG2C, or any combination thereof; and   (b) the chimeric intracellular signaling molecule substantially lacks an extracellular ligand-binding domain.   
     
     
         33 . The population of cells of  claim 32  further comprising a bispecific antibody. 
     
     
         34 . The metabolically enhanced T cell of  claim 1 , wherein the chimeric intracellular signaling molecule further comprises a detectable tag. 
     
     
         35 . The metabolically enhanced T cell of  claim 1 , wherein the hinge and transmembrane domain are: (1) CD8-alpha hinge and CD8alpha transmembrane domain; or (2) a CD8-alpha hinge and a CD28 transmembrane domain. 
     
     
         36 . The metabolically enhanced T cell of  claim 4 , wherein the bispecific antibody is chemically heteroconjugated to a polyclonal antibody specific for a tumor-associated antigen (TAA), and the metabolically enhanced T cell specifically binds the TAA polyclonal antibody. 
     
     
         37 . The metabolically enhanced T cell of  claim 4 , wherein the bispecific antibody comprises an antigen binding domain comprising a first whole immunoglobulin molecule and a second whole IgG immunoglobulin molecule. 
     
     
         38 . The metabolically enhanced T cell of  claim 37 , wherein at least one of the first or second whole immunoglobulin molecules comprises IgG, IgA, or IgD. 
     
     
         39 . The metabolically enhanced T cell of  claim 4 , wherein the metabolically enhanced T cell comprises two or more bispecific antibodies and the T cell displays the two or more bispecific antibodies. 
     
     
         40 . The metabolically enhanced T cell of  claim 39 , wherein the two or more bispecific antibodies comprise a combination of antibodies selected from the group consisting of anti-CD3, anti-IgD Fc, and anti-IgA Fc.