Engineered cells for adoptive cell therapy

Abstract

Provided are engineered cells for adoptive therapy, including NK cells and T cells. Also provided are compositions for engineering and producing the cells, compositions containing the cells, and methods for their administration to subjects. In some embodiments, the cells contain genetically engineered antigen receptors that specifically bind to antigens, such as chimeric antigen receptors (CARs) and costimulatory receptors. In some embodiments, the cells include receptors targeting multiple antigens. In some embodiments, the cells include repression of one or more gene product, for example, by disruption of a gene encoding the gene product. In some embodiments, a gene encoding an antigen recognized by the engineered antigen receptor is disrupted, reducing the likelihood of targeting of the engineered cells. In some embodiments, the antigen recognized by the engineered antigen receptor is related to a tumor antigen recognized by the engineered antigen receptor.

1 . An engineered immune cell comprising:
 a recombinant receptor that specifically binds to a target antigen, wherein the target antigen is a B cell maturation antigen (BCMA), a transmembrane activator and CAML Interactor (TACI) or a B-cell activating factor receptor (BAFF-R); and   a genetic disruption resulting in reduced expression of the target antigen in the engineered immune cell.   
     
     
         2 . The engineered immune cell of  claim 1 , wherein the genetic disruption comprises a disruption in a gene encoding the target antigen. 
     
     
         3 . The engineered immune cell of  claim 1 , wherein the recombinant receptor is a chimeric antigen receptor (CAR) or a T cell receptor (TCR). 
     
     
         4 . A pharmaceutical composition comprising the engineered immune cell of  claim 1  and a pharmaceutically acceptable carrier. 
     
     
         5 . A method of treatment, comprising administering the engineered immune cell of  claim 1  to a subject having a disease or condition. 
     
     
         6 . A method of producing a genetically engineered immune cell, comprising:
 (a) introducing into an immune cell a nucleic acid encoding a recombinant receptor that specifically binds to a target antigen, wherein the target antigen is a B cell maturation antigen (BCMA), a transmembrane activator and CAML Interactor (TACI) or a B-cell activating factor receptor (BAFF-R); and   (b) effecting repression of expression of the target antigen in the immune cell;   thereby producing a genetically engineered immune cell in which expression of the target antigen is reduced,   wherein steps (a) and (b) are carried out simultaneously or sequentially in any order.   
     
     
         7 . The method of  claim 6 , wherein the effecting repression comprises disrupting a gene encoding the target antigen. 
     
     
         8 . The method of  claim 7 , wherein the disrupting comprises disrupting the gene at the DNA level and/or
 the disrupting is not reversible; and/or   the disrupting is not transient.   
     
     
         9 . The method of  claim 7 , wherein the disrupting comprises introducing into the immune cell a DNA binding protein or DNA-binding nucleic acid that specifically recognizes the gene, or a fusion protein comprising a DNA-targeting protein and a nuclease or an RNA-guided nuclease. 
     
     
         10 . The method of  claim 9 , wherein the disrupting comprises introducing into the immune cell a zinc finger nuclease (ZFN), a TAL-effector nuclease (TALEN), or and a CRISPR-Cas9 combination that specifically binds to, recognizes, or hybridizes to the gene encoding the target antigen. 
     
     
         11 . The method of  claim 6 , wherein the recombinant receptor is a T cell receptor (TCR) or a chimeric antigen receptor (CAR). 
     
     
         12 . A method of producing a genetically engineered immune cell, comprising:
 (a) introducing into an immune cell a nucleic acid encoding a recombinant receptor that specifically binds to a target antigen, wherein the target antigen is a B cell maturation antigen (BCMA), a transmembrane activator and CAML Interactor (TACI) or a B-cell activating factor receptor (BAFF-R); and   (b) effecting repression of expression of a related antigen in the immune cell, wherein the recombinant receptor also binds or is suspected of binding to the related antigen, which is related to the target antigen,   thereby producing a genetically engineered immune cell in which expression of the related antigen is reduced,   wherein steps (a) and (b) are carried out simultaneously or sequentially in any order.   
     
     
         13 . The method of  claim 12 , wherein the target antigen is a B cell maturation antigen (BCMA) and the related antigen is a different BCMA family member. 
     
     
         14 . The method of  claim 13 , wherein the different BCMA family member is a transmembrane activator and CAML Interactor (TACI) or a B-cell activating factor receptor (BAFF-R). 
     
     
         15 . The method of  claim 12 , wherein the effecting repression comprises disrupting a gene encoding the target antigen or the related antigen. 
     
     
         16 . The method of  claim 15 , wherein the disrupting comprises disrupting the gene at the DNA level and/or
 the disrupting is not reversible; and/or   the disrupting is not transient.   
     
     
         17 . The method of  claim 15 , wherein the disrupting comprises introducing into the immune cell a DNA binding protein or DNA-binding nucleic acid that specifically binds to or hybridizes to the gene, or a fusion protein comprising a DNA-targeting protein and a nuclease or an RNA-guided nuclease. 
     
     
         18 . The method of  claim 17 , wherein the disrupting comprises introducing into the immune cell a zinc finger nuclease (ZFN), a TAL-effector nuclease (TALEN), or and a CRISPR-Cas9 combination that specifically binds to, recognizes, or hybridizes to the gene encoding the target antigen. 
     
     
         19 . The method of  claim 12 , wherein the recombinant receptor is a T cell receptor (TCR) or a chimeric antigen receptor (CAR). 
     
     
         20 . An engineered immune cell, comprising:
 (a) a first recombinant receptor that specifically binds to a first antigen and is capable of inducing an activating signal to the cell; and   (b) a second recombinant receptor that specifically binds to a second antigen or a second recombinant receptor which is a chimeric costimulatory receptor that specifically binds to a second antigen and is capable of inducing a costimulatory signal to the cell,   wherein the first antigen and the second antigen are distinct and at least one of the first antigen or the second antigen is a B cell maturation antigen (BCMA), a transmembrane activator and CAML Interactor (TACI) or a B-cell activating factor receptor (BAFF-R).   
     
     
         21 . The engineered immune cell of  claim 20 , wherein the other of the first antigen or the second antigen is an antigen expressed in a tumor. 
     
     
         22 . The engineered immune cell of  claim 20 , wherein the first antigen or the second antigen is a BCMA. 
     
     
         23 . The engineered immune cell of  claim 22 , wherein the other of the first antigen or the second antigen is a BCMA-related family member. 
     
     
         24 . The engineered immune cell of  claim 23 , wherein the different BCMA family member is a transmembrane activator and CAML Interactor (TACI) or a B-cell activating factor receptor (BAFF-R). 
     
     
         25 . The engineered immune cell of  claim 22 , wherein the other of the first antigen or the second antigen is a FcRH5, a CS1, a CD38 or a CD138. 
     
     
         26 . The engineered immune cell of  claim 20 , further comprising a genetic disruption in a gene encoding the first antigen, and/or in a gene encoding the second antigen, said genetic disruption resulting in reduced expression of the first and/or second antigen in the engineered immune cell. 
     
     
         27 . The engineered immune cell of  claim 20 , wherein the first recombinant receptor is a T cell receptor (TCR) or a chimeric antigen receptor (CAR). 
     
     
         28 . A pharmaceutical composition comprising the engineered immune cell of  claim 20  and a pharmaceutically acceptable carrier. 
     
     
         29 . A method of treatment, comprising administering the engineered immune cell of  claim 20  to a subject having a disease or condition. 
     
     
         30 . A method of producing a genetically engineered immune cell, the method comprising:
 (a) introducing into an immune cell a nucleic acid encoding a first recombinant receptor that specifically binds to a first antigen; and   (b) introducing into the immune cell a nucleic acid encoding a second recombinant receptor that specifically binds to a second antigen or a second recombinant receptor that is a chimeric costimulatory receptor and specifically binds to a second antigen,   thereby producing the engineered immune cell,   wherein the first antigen and the second antigen are distinct and at least one of the first antigen or the second antigen is a B cell maturation antigen (BCMA), a transmembrane activator and CAML Interactor (TACI) or a B-cell activating factor receptor (BAFF-R) and   steps (a) and (b) are carried out simultaneously or sequentially in any order.   
     
     
         31 . The method of  claim 30 , wherein the other of the first antigen or the second antigen is an antigen expressed in a tumor. 
     
     
         32 . The method of  claim 30 , wherein the first antigen or the second antigen is a BCMA. 
     
     
         33 . The method of  claim 32 , wherein the other of the first antigen or the second antigen is a BCMA-related family member. 
     
     
         34 . The method of  claim 33 , wherein the different BCMA family member is a transmembrane activator and CAML Interactor (TACI) or a B-cell activating factor receptor (BAFF-R). 
     
     
         35 . The method of  claim 33 , wherein the other of the first antigen or the second antigen is a FcRH5, a CS1, a CD38 or a CD138. 
     
     
         36 . The method of  claim 30 , further comprising (c) effecting repression of expression of the first antigen and/or the second antigen in the immune cell, wherein the effecting repression comprises disrupting a gene encoding the first antigen and/or the second antigen. 
     
     
         37 . The method of  claim 36 , wherein the disrupting comprises introducing into the immune cell a DNA binding protein or DNA-binding nucleic acid that specifically binds to or hybridizes to the gene encoding the first antigen and/or the second antigen or introducing a zinc finger nuclease (ZFN), a TAL-effector nuclease (TALEN), or and a CRISPR-Cas9 combination that specifically binds to, recognizes, or hybridizes to the gene encoding the first antigen and/or the second antigen. 
     
     
         38 . A composition or a combination, comprising:
 (a) a first engineered immune cell expressing a first recombinant receptor that specifically binds to a first antigen and not expressing a second recombinant receptor that specifically binds to a second antigen; and   (b) a second engineered immune cell expressing the second recombinant receptor,   wherein the first antigen and the second antigen are distinct and at least one of the first antigen or the second antigen is a B cell maturation antigen (BCMA), a transmembrane activator and CAML Interactor (TACI) or a B-cell activating factor receptor (BAFF-R).   
     
     
         39 . The composition or combination of  claim 38 , wherein the first antigen or the second antigen is BCMA. 
     
     
         40 . The composition or combination of  claim 39 , wherein the other of the first antigen or the second antigen is a BCMA-related family member. 
     
     
         41 . The composition or combination of  claim 40 , wherein the different BCMA family member is a transmembrane activator and CAML Interactor (TACI) or a B-cell activating factor receptor (BAFF-R). 
     
     
         42 . The composition or combination of  claim 39 , wherein the other of the first antigen or the second antigen is a FcRH5, a CS1, a CD38 or a CD138. 
     
     
         43 . A method of treatment, comprising administering the composition or combination of  claim 38  to a subject having a disease or condition.