Enhancing Activity of CAR T Cells by Co-Introducing a Bispecific Antibody

Abstract

The present invention provides compositions and methods for treating cancer in a human. The invention includes administering a T cell, genetically modified to express a chimeric antigen receptor (CAR), a bispecific antibody, or a combination thereof to a subject. The CAR and bispecific antibody of the invention can comprise a human antibody, a humanized antibody, or antigen-binding fragments thereof.

1 . An isolated nucleic acid molecule comprising a nucleic acid sequence encoding a chimeric antigen receptor (CAR), wherein the CAR comprises an antigen binding domain derived from a bispecific antibody, a transmembrane domain, and a CD 3  zeta signaling domain, further wherein the antigen binding domain is selected from the group consisting of a human antibody, a humanized antibody, an antigen binding fragment thereof, and any combination thereof. 
     
     
         2 . The isolated nucleic acid molecule of  claim 1 , wherein the nucleic acid sequence comprises SEQ ID NO:  20 ,  21 , or  22 . 
     
     
         3 . The isolated nucleic acid sequence of  claim 1 , wherein the antigen-binding fragment is a Fab or a scFv. 
     
     
         4 . The isolated nucleic acid molecule of  claim 1 , wherein the antigen binding domain binds to a tumor antigen. 
     
     
         5 . The isolated nucleic acid molecule of  claim 4 , wherein the tumor antigen is associated with a hematologic malignancy. 
     
     
         6 . The isolated nucleic acid molecule of  claim 4 , wherein the tumor antigen is associated with a solid tumor. 
     
     
         7 . The isolated nucleic acid molecule of  claim 4 , wherein the tumor antigen is 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 any combination thereof. 
     
     
         8 . The isolated nucleic acid molecule of  claim 1 , wherein the CAR further comprises a costimulatory signaling region comprising an intracellular domain of a costimulatory molecule selected from the group consisting of CD27, CD28, 4-1BB, OX40, CD30, CD40, PD-1, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83, and any combination thereof. 
     
     
         9 . The isolated nucleic acid molecule of  claim 1 , wherein the nucleic acid sequence of the antigen binding domain derived from a bispecific antibody encodes a bispecific antibody. 
     
     
         10 . The isolated nucleic acid molecule of  claim 9 , wherein the nucleic acid sequence encoding the bispecific antibody comprises the nucleic acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, and any combination thereof. 
     
     
         11 - 47 . (canceled) 
     
     
         48 . An isolated nucleic acid molecule comprising:
 a first nucleic acid sequence encoding a chimeric antigen receptor (CAR), wherein the CAR comprises one or more antigen binding domains of a first bispecific antibody, a transmembrane domain, and a CD3 zeta signaling domain, further wherein each antigen binding domain is selected from the group consisting of a human antibody, a humanized antibody, and an antigen binding fragment thereof; and   a second nucleic acid sequence encoding a second bispecific antibody, wherein the second bispecific antibody does not comprise and is not linked to a transmembrane domain.   
     
     
         49 . The isolated nucleic acid molecule of  claim 48 , wherein the second bispecific antibody comprises a first antigen recognition domain that recognizes the same antigen as the antigen binding domain of the CAR, and a second antigen recognition domain that recognizes CD3. 
     
     
         50 . The isolated nucleic acid molecule of  claim 48 , wherein the first nucleic acid sequence comprises the nucleic acid sequence of SEQ ID NO: 20, 21, or 22. 
     
     
         51 . The isolated nucleic acid molecule of  claim 48 , wherein at least one antigen binding domain of the CAR:
 (a) is an antigen binding fragment; or   (b) binds to a tumor antigen.   
     
     
         52 . The isolated nucleic acid molecule of  claim 51 , wherein the antigen binding fragment is a Fab or a scFv. 
     
     
         53 . The isolated nucleic acid molecule of  claim 51 , wherein the tumor antigen is associated with a hematologic malignancy or a solid tumor. 
     
     
         54 . The isolated nucleic acid molecule of  claim 51 , wherein the tumor antigen is selected from the group consisting of CD19, CD20, CD22, ROR1, mesothelin, CD33, IL3Ra, c-Met, PSMA, Glycolipid F77, EGFRvIII, GD-2, NY-ESO-1, and MAGE A3. 
     
     
         55 . The isolated nucleic acid molecule of  claim 48 , wherein the isolated nucleic acid molecule is an RNA molecule. 
     
     
         56 . The isolated nucleic acid molecule of  claim 48 , wherein the CAR further comprises a costimulatory signaling region comprising an intracellular domain of a costimulatory molecule selected from the group consisting of CD27, CD28, 4-1BB, OX40, CD30, CD40, PD-1, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, CD83, and any combination thereof. 
     
     
         57 . The isolated nucleic acid molecule of  claim 48 , wherein the nucleic acid sequence encoding the one or more antigen binding domains of the first bispecific antibody comprises a nucleic acid sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, and SEQ ID NO: 19.