Use of the cd2 signaling domain in second-generation chimeric antigen receptors

Abstract

The present invention provides compositions and methods for treating cancer in a human. The invention includes relates to administering a genetically modified T cell expressing a CAR having an antigen binding domain, a transmembrane domain, a CD2 signaling domain, and a CD3 zeta signaling domain. The invention also includes incorporating CD2 into the CAR to alter the cytokine production of CAR-T cells in both negative and positive directions.

1 - 5 . (canceled) 
     
     
         6 . An isolated chimeric antigen receptor (CAR) comprising an antigen binding domain, a transmembrane domain, a costimulatory signaling region, and a CD3 zeta signaling domain, wherein:
 (a) the antigen binding domain targets a tumor antigen selected from the group consisting of CD19, CD20, CD22, ROR1, mesothelin, CD33, IL3Ra, c-Met, PSMA, Glycolipid F77, EGFRvIII, GD-2, NY-ESO-1 TCR, MAGE A3 TCR, and MUC1 (CA15-3), and any combination thereof; and   (b) the costimulatory signaling region comprises a CD2 signaling domain.   
     
     
         7 . The isolated CAR of  claim 6 , wherein the antigen binding domain is an antibody or an antigen-binding fragment thereof. 
     
     
         8 . (canceled) 
     
     
         9 . The isolated CAR of  claim 6 , wherein the costimulatory signaling region further comprises the intracellular domain of a costimulatory molecule selected from the group consisting of CD27, CD28, 4-1BB, OX40, CD30, CD40, PD-1, ICOS, lymphocyte function-associated antigen-I (LFA-1), CD7, LIGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83, and any combination thereof. 
     
     
         10 - 28 . (canceled) 
     
     
         29 . A method of generating a persisting population of genetically engineered T cells in a human diagnosed with cancer, the method comprising administering to the human a T cell genetically engineered to express the isolated CAR of  claim 6 , wherein the persisting population of genetically engineered T cells persists in the human for at least one month after administration. 
     
     
         30 . The method of  claim 29 , wherein the persisting population of genetically engineered T cells comprises at least one cell selected from the group consisting of a T cell that was administered to the human, a progeny of a T cell that was administered to the human, and a combination thereof. 
     
     
         31 . The method of  claim 29 , wherein the persisting population of genetically engineered T cells comprises a memory T cell. 
     
     
         32 . The method of  claim 29 , wherein the persisting population of genetically engineered T cells persists in the human for at least three months after administration. 
     
     
         33 . The method of  claim 29 , wherein the persisting population of genetically engineered T cells persists in the human for at least four months, five months, six months, seven months, eight months, nine months, ten months, eleven months, twelve months, two years, or three years after administration. 
     
     
         34 . The method of  claim 29 , wherein the cancer is treated. 
     
     
         35 . A method of expanding a population of genetically engineered T cells in a human diagnosed with cancer, the method comprising administering to the human a T cell genetically engineered to express the isolated CAR of  claim 6 , wherein the administered genetically engineered T cell produces a population of progeny T cells in the human. 
     
     
         36 . The method of  claim 35 , wherein the progeny T cells in the human comprise a memory T cell. 
     
     
         37 . The method of  claim 35 , wherein the T cell is an autologous T cell. 
     
     
         38 . The method of  claim 35 , wherein the population of progeny T cells persists in the human for at least three months after administration. 
     
     
         39 . The method of  claim 35 , wherein the population of progeny T cells persist in the human for at least four months, five months, six months, seven months, eight months, nine months, ten months, eleven months, twelve months, two years, or three years after administration. 
     
     
         40 . A method of modulating the amount of cytokine secreted by a T cell, said method comprising genetically engineering the T cell to express a CAR, wherein the CAR comprises an antigen binding domain, a transmembrane domain, a costimulatory domain region comprising a CD2 signaling domain, and a CD3 zeta signaling domain. 
     
     
         41 . The method of  claim 40 , wherein modulating the amount of cytokine secreted by a T cell reduces the proliferation of T regulatory cells. 
     
     
         42 . A method of reducing the amount of activation-induced calcium influx into a T cell, said method comprising genetically engineering the T cell to express a CAR, wherein the CAR comprises an antigen binding domain, a transmembrane domain, a costimulatory domain region comprising a CD2 signaling domain, and a CD3 zeta signaling domain. 
     
     
         43 . The method of  claim 42 , wherein reducing the amount of activation-induced calcium influx into a T cell prevents the activation-induced cell death of the T cell. 
     
     
         44 . The method of  claim 40 , wherein the CD2 signaling domain is encoded by a nucleic acid sequence comprising nucleotides 996-1346 of SEQ ID NO: 1 
     
     
         45 . The method of  claim 40 , wherein the antigen binding domain targets a tumor antigen selected from the group consisting of CD19, CD20, CD22, ROR1, mesothelin, CD33, IL3Ra, c-Met, PSMA, Glycolipid F77, EGFRvIII, GD-2, NY-ESO-1 TCR, MAGE A3 TCR, and MUC1 (CA15-3), and any combination thereof. 
     
     
         46 . The method of  claim 40 , wherein the nucleic acid encoding the CAR comprises the nucleic acid sequence of SEQ ID NO: 1. 
     
     
         47 . The method of  claim 40 , wherein the CD2 signaling domain increases cytokine production. 
     
     
         48 . The method of  claim 40 , wherein the CD2 signaling domain reduces cytokine production. 
     
     
         49 . The method of  claim 40 , wherein the costimulatory signaling region further comprises the intracellular domain of a costimulatory molecule selected from the group consisting of CD27, CD28, 4-1BB, OX40, CD30, CD40, PD-1, ICOS, lymphocyte function-associated antigen-I (LFA-1), CD7, LIGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83, and any combination thereof. 
     
     
         50 . The method of  claim 40 , wherein the costimulatory signaling region further comprises the costimulatory domain of CD28. 
     
     
         51 . The method of claim  18 , wherein a modified T cell expressing a CAR comprising the CD2 and the CD28 costimulatory domains releases significantly less IL-2 compared to a modified T cell expressing a CAR only comprising the CD28 costimulatory domain. 
     
     
         52 . The method of  claim 42 , wherein the costimulatory signaling region further comprises the intracellular domain of a costimulatory molecule selected from the group consisting of CD27, CD28, 4-1BB, OX40, CD30, CD40, PD-1, ICOS, lymphocyte function-associated antigen-I (LFA-1), CD7, LIGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83, and any combination thereof. 
     
     
         53 . The isolated CAR of  claim 6 , wherein the CD2 signaling domain is encoded by a nucleic acid sequence comprising nucleotides 996-1346 of SEQ ID NO: 1.