Modified Cell Expressing Therapeutic Agent and Uses thereof

Abstract

Compositions and methods for enhancing T cell response which increases the efficacy of CAR T cell therapy for treating cancer are described. Embodiments include a modified cell comprising an isolated nucleic acid comprising a first nucleic acid and a second nucleic acid, the first nucleic acid encoding a chimeric antigen receptor (CAR), the second nucleic acid encoding a therapeutic agent comprising at least one of IFN-γ, IL-2, IL-6, IL-7, IL-15, IL-17, and IL-23. The modified cell expresses and secretes the therapeutic agent.

1 . A method of enhancing cytokine release, the method comprising:
 introducing a polynucleotide encoding IL-12 into a modified T cell comprising a binding molecule binding an antigen;   contacting the modified T cell with a cell comprising the antigen; and   measuring levels of cytokine release of IL-6 and IFNγ of the modified T cells, wherein the levels of cytokine release of IL-6 and IFNγ of the modified T cells are greater than levels of cytokine release of IL-6 and IFNγ of a modified T cell comprising the binding molecule but not the polynucleotide.   
     
     
         2 . The method of  claim 1 , wherein the polynucleotide comprises a promoter comprising a binding site for a transcription modulator that modulates expression and/or secretion of IL-12 in the modified T cell. 
     
     
         3 . The method of  claim 2 , wherein the transcription modulator comprises Hif1a and NFAT, and wherein IL-12 is expressed in response to activation of the modified T cell in hypoxia. 
     
     
         4 . The method of  claim 1 , wherein the binding molecule is a Chimeric Antigen Receptor (CAR). 
     
     
         5 . The method of  claim 4 , wherein the CAR binds tMUC 1, PRLR, CLCA1, MUC12, GUCY2C, GPR35, CR1L, MUC 17, TMPRSS11B, MUC21, TMPRSS11E, CD207, SLC30A8, CFC1, SLC12A3, SSTR1, GPR27, FZD10, TSHR, SIGLEC15, SLC6A3, KISS1R, QRFPR, GPR119, CLDN6, UPK2, ADAM12, SLC45A3, ACPP, MUC21, MUC16, MS4A12, ALPP, CEA, EphA2, FAP, GPC3, IL13-Ra2, Mesothelin, PSMA, ROR1, VEGFR-II, GD2, FR-α, ErbB2, EpCAM, EGFRvIII, PSCA, or EGFR. 
     
     
         6 . The method of  claim 4 , wherein the CAR comprises an extracellular domain, a transmembrane domain, and an intracellular domain, the extracellular domain binding an antigen. 
     
     
         7 . The method of  claim 6 , wherein the intracellular domain comprises a co-stimulatory domain that comprises an intracellular domain of a co-stimulatory molecule comprising CD27, CD28, 4-1BB, OX40, CD30, CD40, PD-1, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, or a combination thereof. 
     
     
         8 . The method of  claim 6 , wherein the antigen comprises Epidermal growth factor receptor (EGFR), Variant III of the epidermal growth factor receptor (EGFRvIII), Human epidermal growth factor receptor 2 (HER2), Mesothelin (MSLN), Prostate-specific membrane antigen (PSMA), Carcinoembryonic antigen (CEA), Disialoganglioside 2 (GD2), Interleukin-13Ra2 (IL13Rα2), Glypican-3 (GPC3), Carbonic anhydrase IX (CAIX), L1 cell adhesion molecule (L1-CAM), Cancer antigen 125 (CA125), Cluster of differentiation 133 (CD133), Fibroblast activation protein (FAP), Cancer/testis antigen 1B (CTAG1B), Mucin 1 (MUC1), Folate receptor-α (FR-α), CD19, FZD10, TSHR, PRLR, Muc 17, GUCY2C, CD207, CD3, CD5, B-Cell Maturation Antigen (BCMA), or CD4. 
     
     
         9 . The method of  claim 1 , wherein the polynucleotide is in a recombinant DNA construct, in an mRNA, or in a viral vector. 
     
     
         10 . The method of  claim 1 , wherein expression of the polynucleotide is regulated by SynNotch polypeptide. 
     
     
         11 . The method of  claim 1 , wherein the binding molecule is a modified TCR. 
     
     
         12 . The method of  claim 11 , wherein the TCR is derived from spontaneously occurring tumor-specific T cells in patients. 
     
     
         13 . The method of  claim 11 , wherein the TCR binds a tumor antigen. 
     
     
         14 . The method of  claim 13 , wherein the tumor antigen comprises CEA, gp100, MART-1, p53, MAGE-A3, or NY-ESO-1, or the TCR comprises TCRγ and TCRδ Chains or TCRα and TCRβ chains, or a combination thereof. 
     
     
         15 . The method of  claim 1 , wherein the modified T cell further comprises an additional CAR binding an additional antigen of a white blood cell (WBC), and the additional antigen is different from the antigen. 
     
     
         16 . The method of  claim 1 , wherein the modified T cell further comprises a dominant negative form of PD-1. 
     
     
         17 . The method of  claim 1 , wherein the modified T cell has a reduced expression of endogenous TRAC gene. 
     
     
         18 . The method of  claim 1 , wherein the modified T cell is a T cell derived from a primary human T cell isolated from a human donor. 
     
     
         19 . The method of  claim 1 , wherein expression of IL-12 is regulated by an inducible gene expression system. 
     
     
         20 . The method of  claim 1 , wherein the binding molecule is a bispecific CAR binding two different antigens.