Immune cells expressing engineered antigen receptors

Abstract

Provided herein are immune cells expressing antigenic receptors, such as a chimeric antigen receptor and a T cell receptor. Further provided herein are methods of treating immune-related disorder by administering the antigen-specific immune cells.

What is claimed is: 
     
         1 . An immune cell engineered to express human IL-15 (hIL-15) and at least two antigen receptors, wherein the at least two antigen receptors comprise a chimeric antigen receptor (CAR) and/or a T cell receptor (TCR). 
     
     
         2 . The immune cell of  claim 1 , wherein the immune cell is engineered to express hIL-15, the CAR, and the TCR. 
     
     
         3 . The immune cell of  claim 1 , wherein the immune cell is engineered to express hIL-15 and two CARs. 
     
     
         4 . The immune cell of  claim 1 , wherein the immune cell is engineered to express hIL-15 and two TCRs. 
     
     
         5 . The immune cells of  claim 1 , wherein the immune cell is engineered to express 3, 4, or 5 antigen receptors. 
     
     
         6 . The immune cell of any one of  claims 1 - 5 , wherein the immune cell is further defined as a T cell, peripheral blood lymphocyte, NK cell, invariant NK cell, NKT cell, or stem cell. 
     
     
         7 . The immune cell of any one of  claims 1 - 5 , wherein the immune cell is a T cell. 
     
     
         8 . The immune cell of any one of  claims 1 - 5 , wherein the immune cell is an NK cell. 
     
     
         9 . The immune cell of  claim 6 , wherein the stem cell is a mesenchymal stem cell (MSC) or an induced pluripotent stem (iPS) cell. 
     
     
         10 . The immune cell of  claim 1 , wherein the immune cell is derived from an iPS cell. 
     
     
         11 . The immune cell of  claim 7 , wherein the T cell is a CD8 +  cell, CD4 +  T cell, or gamma-delta cell. 
     
     
         12 . The immune cell of  claim 7 , wherein the T cell is a cytotoxic T lymphocyte (CTL). 
     
     
         13 . The immune cell of  claim 6 , wherein the immune cell is allogeneic. 
     
     
         14 . The immune cell of  claim 6 , wherein the immune cell is autologous. 
     
     
         15 . The immune cell of any of  claims 1 - 13 , wherein the immune cell is engineered to express one or more additional cytokines. 
     
     
         16 . The immune cell of  claim 15 , wherein the one or more additional cytokines are IL-21 and/or IL-2. 
     
     
         17 . The immune cell of any one of  claims 1 - 13 , wherein the immune cell is engineered to have essentially no expression of glucocorticoid receptor, TGFβ receptor, and/or CISH. 
     
     
         18 . The immune cell of  claim 17 , wherein said immune cell is engineered using one or more guide RNAs and a Cas9 enzyme. 
     
     
         19 . The immune cell of  claim 18 , wherein the one or more guide RNAs comprise SEQ ID NOs. 1-2. 
     
     
         20 . The immune cell of  claim 18 , wherein the one or more guide RNAs comprise SEQ ID NOs. 3-4. 
     
     
         21 . The immune cell of  claim 17 , wherein the TGFβ receptor is further defined as TGFβ-RII. 
     
     
         22 . The immune cell of any one of  claims 1 - 13 , wherein the immune cell is isolated from peripheral blood, cord blood, or bone marrow. 
     
     
         23 . The immune cell of any one of  claims 1 - 13 , wherein the immune cell is isolated from cord blood. 
     
     
         24 . The immune cell of  claim 23 , wherein the cord blood is pooled from 2 or more individual cord blood units. 
     
     
         25 . The immune cell of any one of  claims 1 - 13 , wherein the immune cell further expresses a suicide gene. 
     
     
         26 . The immune cell of  claim 25 , wherein the suicide gene is CD20, CD52, EGFRv3, or inducible caspase 9. 
     
     
         27 . The immune cell of  claim 25 , wherein the suicide gene is inducible caspase 9. 
     
     
         28 . The immune cell of  claim 1 , wherein DNA encoding the at least two antigen receptors is integrated into the genome of the cell. 
     
     
         29 . The immune cell of  claim 1 , wherein DNA encoding the CAR and/or TCR is integrated into the genome of the cell. 
     
     
         30 . The immune cell of  claim 1 , wherein the at least two antigen receptors comprise antigen binding regions selected from the group consisting of F(ab′)2, Fab′, Fab, Fv, and scFv. 
     
     
         31 . The immune cell of  claim 30 , wherein the antigen binding regions of the at least two antigen receptors bind one or more tumor associated antigens. 
     
     
         32 . The immune cell of  claim 31 , wherein the tumor associated antigens are CD19, CD319/CS1, ROR1, CD20, carcinoembryonic antigen, alphafetoprotein, CA-125, MUC-1, epithelial tumor antigen, melanoma-associated antigen, mutated p53, mutated ras, HER2/Neu, ERBB2, folate binding protein, HIV-1 envelope glycoprotein gp120, HIV-1 envelope glycoprotein gp41, GD2, CD123, CD23, CD30, CD56, c-Met, mesothelin, GD3, HERV-K, IL-11Ralpha, kappa chain, lambda chain, CSPG4, ERBB2, WT-1, EGFRvIII, TRAIL/DR4, and/or VEGFR2. 
     
     
         33 . The immune cell of  claim 30 , wherein the antigen binding region of a first antigen receptor is distinct from the antigen binding region of a second antigen receptor. 
     
     
         34 . The immune cell of  claim 32 , wherein the antigen binding region of the first antigen receptor binds to a first antigen and the antigen binding region of the second antigen receptor binds to a second antigen. 
     
     
         35 . The immune cell of  claim 34 , wherein first antigen is EGFRvIII and the second antigen is NY-ESO. 
     
     
         36 . The immune cell of  claim 34 , wherein first antigen is HER2/Neu and the second antigen is MUC-1. 
     
     
         37 . The immune cell of  claim 34 , wherein first antigen is CA-125 and the second antigen is MUC-1. 
     
     
         38 . The immune cell of  claim 34 , wherein first antigen is CA-125 and the second antigen is WT-1. 
     
     
         39 . The immune cell of  claim 34 , wherein first antigen is EGFRvIII and the second antigen is Mage-A3, Mage-A4, or Mage-A10. 
     
     
         40 . The immune cell of  claim 34 , wherein first antigen is EGFRvIII and the second antigen is TRAIL/DR4. 
     
     
         41 . The immune cell of  claim 34 , wherein first antigen is CEA-CAR and the second antigen is Mage-A3-TCR, Mage-A4-TCR or Mage-A10. 
     
     
         42 . The immune cell of  claim 34 , wherein first antigen is HER2/Neu, CEA-CAR, and/or CA-125, EGFRvIII and the second antigen is MUC-1, WT-1, TRAIL/DR4Mage-A3-TCR, Mage-A4-TCR and/or Mage-A10. 
     
     
         43 . The immune cell of any one of  claims 1 - 13 , wherein the at least two antigen receptors comprise one or more intracellular signaling domains. 
     
     
         44 . The immune cell of  claim 42 , wherein the one or more intracellular signaling domains are T-lymphocyte activation domains. 
     
     
         45 . The immune cell of  claim 42 , wherein the one or more intracellular signaling domains comprise CD3ξ, CD28, OX40/CD134, 4-1BB/CD137, FcεRIγ, ICOS/CD278, ILRB/CD122, IL-2RG/CD132, DAP12, CD70, CD40, or a combination thereof. 
     
     
         46 . The immune cell of  claim 42 , wherein the one or more intracellular signaling domains comprise CD3ξ, CD28, 4-1BB-L, and/or DAP12. 
     
     
         47 . The immune cell of  claim 1 , wherein the at least two antigen receptors comprise one or more transmembrane domains. 
     
     
         48 . The immune cell of  claim 47 , wherein the one or transmembrane domains comprise CD28 transmembrane domain, IgG4Fc hinge, Fc regions, CD4 transmembrane domain, the CD3ξ transmembrane domain, cysteine mutated human CD3ξ domain, CD16 transmembrane domain, CD8 transmembrane domain, and/or erythropoietin receptor transmembrane domain. 
     
     
         49 . A pharmaceutical composition comprising an effective amount of an immune cell of any one of  claims 1 - 48 . 
     
     
         50 . A composition comprising an effective amount of an immune cell of an immune cell of any one of  claims 1 - 48  for the treatment of an immune-related disorder in a subject. 
     
     
         51 . The use of a composition comprising an effective amount of an immune cell of an immune cell of any one of  claims 1 - 48  for the treatment of an immune-related disorder in a subject. 
     
     
         52 . A method of treating an immune-related disorder in a subject comprising administering an effective amount of immune cells of any one of  claims 1 - 48  to the subject. 
     
     
         53 . The method of  claim 52 , wherein the immune-related disorder is a cancer, autoimmune disorder, graft versus host disease, allograft rejection, or inflammatory condition. 
     
     
         54 . The method of  claim 52 , wherein the immune-related disorder is an inflammatory condition and the immune cells have essentially no expression of glucocorticoid receptor. 
     
     
         55 . The method of  claim 54 , wherein the subject has been or is being administered a steroid therapy. 
     
     
         56 . The method of  claim 52 , wherein the immune cells are autologous. 
     
     
         57 . The method of  claim 52 , wherein the immune cells are allogeneic. 
     
     
         58 . The method of  claim 52 , wherein the immune-related disorder is a cancer. 
     
     
         59 . The method of  claim 58 , wherein the cancer is a solid cancer or a hematologic malignancy. 
     
     
         60 . The method of  claim 58 , wherein the cancer is ovarian cancer and the immune cells have antigenic specificity for MUC-1, CA-125, and/or WT-1. 
     
     
         61 . The method of  claim 58 , wherein the cancer is lung cancer and the immune cells have antigenic specificity for NY-ESO, EGFR-vIII, Mage-A3, Mage-A4, Mage-A10, and/or TRAIL/DR4. 
     
     
         62 . The method of  claim 58 , wherein the cancer is pancreatic cancer or colon cancer and the immune cells have antigenic specificity for Mage-A3, Mage-A4, Mage-A10, and/or CEA. 
     
     
         63 . The method of  claim 58 , wherein the cancer is breast cancer and the immune cells have antigenic specificity for MUC-1 and HER2/Neu. 
     
     
         64 . The method of  claim 58 , wherein the cancer is glioblastoma and the immune cells have antigenic specificity for Mage-A3, Mage-A4, Mage-A10v, and/or EGFRvIII. 
     
     
         65 . The method of  claim 58 , wherein the cancer is sarcoma and the immune cells have antigenic specificity for NY-ESO and EGFR-vIII. 
     
     
         66 . The method of  claim 52 , further comprising administering at least a second therapeutic agent. 
     
     
         67 . The method of  claim 66 , wherein the at least a second therapeutic agent comprises chemotherapy, immunotherapy, surgery, radiotherapy, or biotherapy. 
     
     
         68 . The method of  claim 66 , wherein the immune cells and/or the at least a second therapeutic agent are administered intravenously, intraperitoneally, intratracheally, intratumorally, intramuscularly, endoscopically, intralesionally, percutaneously, subcutaneously, regionally, or by direct injection or perfusion.