Compositions and methods for t cell engineering

Abstract

The present disclosure relates to an engineered immune cell and use thereof. The present disclosure provides an engineered immune cell comprising a CAR or engineered TCR, which CAR or engineered TCR can comprise a first antigen binding domain and a second antigen binding domain. The engineered immune cells of the present disclosure, when administered into a subject, can inhibit the host immune cells such as T cells and/or NK cells and enhance the survival and persistence of the engineered immune cells in vivo, thereby exhibiting more effective tumor killing activity.

What is claimed is: 
     
         1 . An engineered immune cell, comprising:
 (i) one or more chimeric polypeptide receptors (CPRs) comprising a binding moiety, wherein the binding moiety comprises:
 a first antigen binding domain exhibiting specific binding to an immune cell antigen, wherein, upon engaging the immune cell antigen with the first antigen binding domain, a subject's immune response toward the engineered immune cell is reduced; and 
 a second antigen binding domain exhibiting specific binding to a disease-associated antigen, wherein the disease-associated antigen and the immune cell antigen are different, 
 wherein each CPR of the one or more CPRs further comprises a transmembrane domain and an intracellular signaling domain; and 
   (ii) a heterologous enhancer moiety capable of enhancing one or more activities of the engineered immune cell,   wherein an endogenous T cell receptor (TCR) of the engineered immune cell is inactivated, and   wherein the engineered immune cell exhibits:
 (1) enhanced degree of persistence by remaining viable for at least about 20 days; 
 (2) enhanced degree of expansion by at least about 10-fold within 15 days; and/or 
 (3) enhanced cytotoxicity against a target cell comprising the disease-associated antigen, as compared to that of a control immune cell that comprises the one or more CPRs and does not comprise the heterologous enhancer moiety. 
   
     
     
         2 . The engineered immune cell of  claim 1 , wherein the engineered immune cell exhibits two or more of (1) the enhanced degree of persistence, (2) the enhanced degree of expansion, and (3) the enhanced cytotoxicity against the target cell. 
     
     
         3 . The engineered immune cell of  claim 1 , wherein (1), (2), and/or (3) is measured in absence of any exogenous enhancer moiety. 
     
     
         4 . The engineered immune cell of  claim 1 , wherein the engineered immune cell exhibits (1) the enhanced degree of persistence and/or (2) the enhanced degree of expansion, in a presence of the disease-associated antigen. 
     
     
         5 . The engineered immune cell of  claim 1 , wherein the engineered immune cell exhibits (1) the enhanced degree of persistence and/or (2) the enhanced degree of expansion, as compared to that of the control immune cell without the heterologous enhancer moiety. 
     
     
         6 . The engineered immune cell of  claim 1 , wherein the enhanced cytotoxicity against the target cell is measured in vivo. 
     
     
         7 . The engineered immune cell of  claim 1 , wherein the heterologous enhancer moiety enhances a cytokine-associated signaling pathway in the engineered immune cell. 
     
     
         8 . The engineered immune cell of  claim 7 , wherein the cytokine is selected from the group consisting of IL-2, IL-3, IL-4, IL-6, IL-7, IL-8, IL-10, IL-11, IL-12, IL-15, IL-17, IL-18, IL-21, IL-23, PD-1, PD-L1, CD122, CSFIR, CTAL-4, TIM-3, CCL21, CCL19, and TGFR beta. 
     
     
         9 . The engineered immune cell of  claim 7 , wherein heterologous enhancer moiety comprises at least a portion of a receptor of the cytokine. 
     
     
         10 . The engineered immune cell of  claim 1 , wherein the one or more CPRs are (a) one or more chimeric antigen receptors (CARs) or (b) one or more engineered T cell receptors (TCRs). 
     
     
         11 . The engineered immune cell of  claim 1 , wherein an endogenous gene encoding the immune cell antigen is inactivated in the engineered immune cell. 
     
     
         12 . The engineered immune cell of  claim 1 , wherein the engineered immune cell further exhibits enhanced expression of endogenous IL-2 and/or endogenous IFN-γ, as compared to that of the control immune cell without the heterologous enhancer moiety. 
     
     
         13 . The engineered immune cell of  claim 1 , wherein a CPR of the one or more CPRs comprises both the first antigen binding domain and the second antigen binding domain. 
     
     
         14 . The engineered immune cell of  claim 1 , wherein (a) a CPR of the one or more CPRs comprises the first antigen binding domain and does not comprise the second antigen binding domain and (b) an additional CPR of the one or more CPRs comprises the second antigen binding domain and does not comprise the first antigen binding domain. 
     
     
         15 . The engineered immune cell of  claim 1 , wherein the immune cell is an NK cell, a T cell, a monocyte, a macrophage, or a granulocyte. 
     
     
         16 . The engineered immune cell of  claim 1 , wherein the engineered immune cell is allogeneic to the subject. 
     
     
         17 . The engineered immune cell of  claim 1 , wherein the immune cell antigen is selected from the group consisting of CD2, CD3, CD4, CD5, CD7, CD8, CD16a, CD16b, CD25, CD27, CD28, CD30, CD38, CD45, CD48, CD50, CD52, CD56, CD57, CD62L, CD69, CD94, CD100, CD102, CD122, CD127, CD132, CD137, CD160, CD161, CD178, CD218, CD226, CD244, CD159a (NKG2A), CD159c (NKG2C), NKG2E, CD279, CD314 (NKG2D), CD305, CD335 (NKP46), CD337, CD319 (CS1), TCRα, TCRβ, and SLAMF7. 
     
     
         18 . The engineered immune cell of  claim 1 , wherein the disease associated antigen is selected from the group consisting of CD2, CD3, CD4, CD5, CD7, CD8, CD19, CD20, CD22, CD25, CD28, CD30, CD33, CD38, CD40, CD44V6, CD47, CD52, CD56, CD57, CD58, CD79b, CD80, CD86, CD81, CD123, CD133, CD137, CD151, CD171, CD276, CLL1, B7H4, BCMA, VEGFR-2, EGFR, GPC3, PMSA, CEACAM6, c-Met, EGFRvIII, ErbB2/HER2, ErbB3, HER-2, HER3, ErbB4/HER-4, EphA2, IGF1R, GD2, O-acetyl GD2, O-acetyl GD3, GHRHR, GHR, Flt1, KDR, Flt4, Flt3, CEA, CA125, CTLA-4, GITR, BTLA, TGFBR1, TGFBR2, TGFBR1, IL6R, gp130, Lewis, TNFR1, TNFR2, PD1, PD-L1, PD-L2, PSCA, HVEM, MAGE-A, MSLN, NY-ESO-1, PSMA, RANK, RORl, TNFRSF4, TWEAK-R, LTPR, LIFRP, LRP5, MUC1, MUC16, TCRα, TCRβ, TLR7, TLR9, PTCH1, WT-1, Robol, Frizzled, OX40, Notch-I-4, APRIL, CS1, MAGE3, Claudin 18.2, Folate receptor α, Folate receptor β, GPC2, CD70, BAFF-R, and TROP-2. 
     
     
         19 . The engineered immune cell of  claim 1 , wherein the immune cell antigen and the disease-associated antigen are, respectively, (a) CD7 and CD19, (b) CD2 and CD19, or (c) TCR and CD19. 
     
     
         20 . The engineered immune cell of  claim 1 , wherein an endogenous gene encoding a subunit of the endogenous TCR is modified, and wherein the gene is selected from the group consisting of TCRα, TCRβ, CD3ε, CD3δ, CD3γ, and CD3ζ. 
     
     
         21 . The engineered immune cell of  claim 1 , wherein the engineered immune cell is derived from an immune cell that is (a) obtained from peripheral blood or cord blood or (b) derived from a stem cell. 
     
     
         22 . A method of treatment of a subject in need thereof, comprising administering to the subject a composition comprising an engineered immune cell, wherein the engineered immune cell comprises:
 (i) one or more chimeric polypeptide receptors (CPRs) comprising a binding moiety, wherein the binding moiety comprises:
 a first antigen binding domain exhibiting specific binding to an immune cell antigen, wherein, upon engaging the immune cell antigen with the first antigen binding domain, a subject's immune response toward the engineered immune cell is reduced, and 
 a second antigen binding domain exhibiting specific binding to a disease-associated antigen, wherein the disease-associated antigen and the immune cell antigen are different, 
 wherein each CPR of the one or more CPRs further comprises a transmembrane domain and an intracellular signaling domain, 
   (ii) a heterologous enhancer moiety capable of enhancing one or more activities of the engineered immune cell,   wherein an endogenous T cell receptor (TCR) of the engineered immune cell is inactivated, and   wherein the engineered immune cell exhibits
 (1) enhanced degree of persistence by remaining viable for at least about 20 days; 
 (2) enhanced degree of expansion by at least about 10-fold within 15 days; and/or 
 (3) enhanced cytotoxicity against a target cell comprising the disease-associated antigen as compared to that of a control immune cell that comprises the one or more CPRs and does not comprise the heterologous enhancer moiety. 
   
     
     
         23 . The method of  claim 22 , wherein the engineered immune cell exhibits two or more of (1) the enhanced degree of persistence, (2) the enhanced degree of expansion, and (3) the enhanced cytotoxicity against the target cell. 
     
     
         24 . The method of  claim 22 , wherein (1), (2), and/or (3) is measured in absence of any exogenous enhancer moiety. 
     
     
         25 . The method of  claim 22 , wherein the heterologous enhancer moiety enhances a cytokine-associated signaling pathway in the engineered immune cell. 
     
     
         26 . The method of  claim 22 , wherein the immune cell antigen and the disease associated antigen are, respectively, (a) CD7 and CD19, (b) CD2 and CD19, or (c) TCR and CD19.