Chimeric antigen receptors containing a chlorotoxin domain

Abstract

Chimeric transmembrane immunoreceptors (CAR) which include an extracellular domain that includes chlorotoxin or a related toxin, or a variant of chlorotoxin or a related toxin, that binds to human glioma or other human tumor cells, a transmembrane region, a costimulatory domain and an intracellular signaling domain are described.

1 .- 32 . (canceled) 
     
     
         33 . A method for treating a malignant glioma comprising administering a therapeutically effective dose of T cells expressing a chimeric antigen receptor, wherein the chimeric antigen receptor comprises: chlorotoxin; a spacer domain, a transmembrane domain selected from: a CD4 transmembrane domain, a CD8 transmembrane domain, a CD28 transmembrane domain, and a CD3ζ transmembrane domain; a costimulatory domain; and CD3 ζ signaling domain. 
     
     
         34 . The method of  claim 33 , wherein the chlorotoxin comprises the amino acid sequence of SEQ ID NO: 1. 
     
     
         35 . The method of  claim 33 , wherein the spacer region comprises 5-300 amino acids. 
     
     
         36 . The method of  claim 33 , wherein the spacer region comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 2-12. 
     
     
         37 . The method of  claim 33 , wherein the spacer comprises an IgG hinge region. 
     
     
         38 . The method of  claim 33 , wherein the chimeric antigen receptor comprising an amino acid sequence: selected from the group consisting of: an amino acid sequence that is identical to SEQ ID NO: 26 or differs from SEQ ID NO: 26 by no more than 5 single amino acid substitutions; an amino acid sequence that is identical to SEQ ID NO: 27 or differs from SEQ ID NO: 27 by no more than 5 single amino acid substitutions; an amino acid sequence that is identical to SEQ ID NO: 28 or differs from SEQ ID NO: 28 by no more than 5 single amino acid substitutions; an amino acid sequence that is identical to SEQ ID NO: 29 or differs from SEQ ID NO: 29 no more than 5 single amino acid substitutions; an amino acid sequence that is identical to SEQ ID NO: 30 or differs from SEQ ID NO: 30 by no more than 5 single amino acid substitutions; an amino acid sequence that is identical to SEQ ID NO: 31 or differs from SEQ ID NO: 31 by no more than 5 single amino acid substitutions; an amino acid sequence that is identical to SEQ ID NO: 32 or differs from SEQ ID NO: 32 by no more than 5 single amino acid substitutions; an amino acid sequence that is identical to SEQ ID NO: 33 or differs from SEQ ID NO: 33 by no more than 5 single amino acid substitutions; an amino acid sequence that is identical to SEQ ID NO: 34 or differs from SEQ ID NO: 34 by no more than 5 single amino acid substitutions; an amino acid sequence that is identical to SEQ ID NO: 35 or differs from SEQ ID NO: 35 by no more than 5 single amino acid substitutions; an amino acid sequence that is identical to SEQ ID NO: 36 or differs from SEQ ID NO: 36 by no more than 5 single amino acid substitutions; an amino acid sequence that is identical to SEQ ID NO: 37 or differs from SEQ ID NO: 37 by no more than 5 single amino acid substitutions; an amino acid sequence that is identical to SEQ ID NO: 38 or differs from SEQ ID NO: 38 by no more than 5 single amino acid substitutions; an amino acid sequence that is identical to SEQ ID NO: 39 or differs from SEQ ID NO: 39 by no more than 5 single amino acid substitutions; and an amino acid sequence that is identical to SEQ ID NO: 40 or differs from SEQ ID NO: 41 by no more than 5 single amino acid substitutions. 
     
     
         39 . The method of  claim 33 , wherein the chimeric antigen receptor consists of an amino acid sequence: selected from the group consisting of: an amino acid sequence that is identical to SEQ ID NO: 26 or differs from SEQ ID NO: 26 by no more than 5 single amino acid substitutions; an amino acid sequence that is identical to SEQ ID NO: 27 or differs from SEQ ID NO: 27 by no more than 5 single amino acid substitutions; an amino acid sequence that is identical to SEQ ID NO: 28 or differs from SEQ ID NO: 28 by no more than 5 single amino acid substitutions; an amino acid sequence that is identical to SEQ ID NO: 29 or differs from SEQ ID NO: 29 no more than 5 single amino acid substitutions; an amino acid sequence that is identical to SEQ ID NO: 30 or differs from SEQ ID NO: 30 by no more than 5 single amino acid substitutions; an amino acid sequence that is identical to SEQ ID NO: 31 or differs from SEQ ID NO: 31 by no more than 5 single amino acid substitutions; an amino acid sequence that is identical to SEQ ID NO: 32 or differs from SEQ ID NO: 32 by no more than 5 single amino acid substitutions; an amino acid sequence that is identical to SEQ ID NO: 33 or differs from SEQ ID NO: 33 by no more than 5 single amino acid substitutions; an amino acid sequence that is identical to SEQ ID NO: 34 or differs from SEQ ID NO: 34 by no more than 5 single amino acid substitutions; an amino acid sequence that is identical to SEQ ID NO: 35 or differs from SEQ ID NO: 35 by no more than 5 single amino acid substitutions; an amino acid sequence that is identical to SEQ ID NO: 36 or differs from SEQ ID NO: 36 by no more than 5 single amino acid substitutions; an amino acid sequence that is identical to SEQ ID NO: 37 or differs from SEQ ID NO: 37 by no more than 5 single amino acid substitutions; an amino acid sequence that is identical to SEQ ID NO: 38 or differs from SEQ ID NO: 38 by no more than 5 single amino acid substitutions; an amino acid sequence that is identical to SEQ ID NO: 39 or differs from SEQ ID NO: 39 by no more than 5 single amino acid substitutions; and an amino acid sequence that is identical to SEQ ID NO: 40 or differs from SEQ ID NO: 41 by no more than 5 single amino acid substitutions. 
     
     
         40 . The method of  claim 33 , wherein the chimeric antigen receptor comprises an amino acid sequence selected from SEQ ID NOs: 26-55. 
     
     
         41 . The method of  claim 33 , wherein the chimeric antigen receptor consists of an amino acid sequence selected from SEQ ID NOs: 26-55. 
     
     
         42 . The method of  claim 33 , wherein the malignant glioma is anaplastic astrocytoma. 
     
     
         43 . The method of  claim 33 , wherein the malignant glioma is glioblastoma. 
     
     
         44 . The method of  claim 33 , wherein the malignant glioma has reoccurred subsequent to a prior treatment for malignant glioma.