Chimeric antigen receptors based on alternative signal 1 domains

Abstract

Described herein are methods for producing and utilizing an alternative signal 1 domain to construct an optimally signaling CAR. Alternative signal 1 domains of the present technology are based on alternatives to CD3ζ, including mutated ITAMs from CD3ζ (which contains 3 IT AM motifs), truncations of CD3ζ, and alternative splice variants known as CD3s, CD3 theta, and artificial constructs engineered to express fusions between CD3s or CD30 and CD3ζ. CAR polypeptides comprising alternative signal 1 domains are utilized to engineer CAR T cells. Further, this technology related to methods of treating cancer by administering to a subject in need thereof CAR T cells comprising alternative signal 1 domains.

1 . A chimeric antigen receptor (CAR) polypeptide comprising:
 a) an extracellular domain comprising a target-binding sequence, wherein the extracellular domain does not bind to HER2;   b) a transmembrane domain;   c) a co-stimulatory domain; and   d) a T cell intracellular signaling domain comprising a mutation of a tyrosine residue in an immunoreceptor tyrosine based activation motif (ITAM) II.   
     
     
         2 . The CAR polypeptide of  claim 1 , wherein the transmembrane domain is the transmembrane domain from is CD8 or 4-1BB. 
     
     
         3 . The CAR polypeptide of  claim 1 , wherein the co-stimulatory domain is the co-stimulatory domain of 4-1BB. 
     
     
         4 . The CAR polypeptide of  claim 1 , wherein the intracellular signaling domain comprises a CD3ζ ITAM3 sequence of SEQ ID NO: 33. 
     
     
         5 .- 6 . (canceled) 
     
     
         7 . The CAR polypeptide of  claim 1 , wherein the target-binding sequence comprises a ligand of the target or an antibody reagent that specifically binds the target. 
     
     
         8 . The CAR polypeptide of  claim 1 , wherein the target is B cell maturation antigen (BCMA). 
     
     
         9 . The CAR polypeptide of  claim 1 , wherein the target is CD37. 
     
     
         10 . The CAR polypeptide of  claim 1 , wherein the antibody reagent comprises an scFv. 
     
     
         11 .- 23 . (canceled) 
     
     
         24 . A mammalian cell comprising:
 a) the CAR polypeptide of  claim 1 ; or   b) a nucleic acid encoding any of the CAR polypeptides of  claim 1 .   
     
     
         25 . The cell of  claim 24 , wherein the cell is a T cell. 
     
     
         26 . The cell of  claim 24 , wherein the cell is a human cell. 
     
     
         27 . The cell of  claim 24 , wherein the cell is obtained from an individual having or diagnosed as having cancer, a plasma cell disorder, or autoimmune disease. 
     
     
         28 . (canceled) 
     
     
         29 . A method of treating cancer, a plasma cell disorder, or an autoimmune disease in a subject in need thereof, the method comprising:
 a) administering the cell of  claim 24  to the subject.   
     
     
         30 . The method of  claim 29 , wherein the cancer is a CD37+ or BCMA+ cancer. 
     
     
         31 . The method of  claim 29 , wherein the CD37+ cancer is lymphoma or leukemia. 
     
     
         32 . The method of  claim 31 , wherein the lymphoma is B-cell Non-Hodgkin Lymphoma (NHL), mantle cell lymphoma, Burkitt's lymphoma, B cell lymphoblastic lymphoma or T cell lymphoma. 
     
     
         33 .- 35 . (canceled) 
     
     
         36 . A method of treating cancer, a plasma cell disorder, or an autoimmune disease in a subject in need thereof, the method comprising administering a cell of  claim 24  to the subject, wherein the cell comprises CAR comprising an extracellular domain comprising a CD37-binding sequence; wherein the subject is non-responsive to anti-CD19 and/or anti-CD20 therapy. 
     
     
         37 . (canceled) 
     
     
         38 . A method of treating cancer, a plasma cell disorder, or an autoimmune disease in a subject in need thereof, the method comprising:
 administering a cell of  claim 24  to the subject, wherein the cell comprises CAR comprising an extracellular domain comprising a CD37-binding sequence; wherein the subject is non-responsive to anti-CD19 and/or anti-CD20 therapy.   
     
     
         39 . (canceled) 
     
     
         40 . A method of treating cancer, a plasma cell disorder, or an autoimmune disease in a subject in need thereof, the method comprising administering a cell of  claim 24  to the subject, wherein the cell comprises CAR comprising an extracellular domain comprising a CD37-binding sequence; wherein the subject is concurrently administered an anti-CD19 and/or anti-CD20 therapy. 
     
     
         41 . A composition comprising the CAR polypeptide of  claim 1  formulated for the treatment of cancer. 
     
     
         42 .- 43 . (canceled)