Cd33 specific chimeric antigen receptors for cancer immunotherapy

Abstract

The present invention relates to Chimeric Antigen Receptors (CAR) that are recombinant chimeric proteins able to redirect immune cell specificity and reactivity toward selected membrane antigens, and more particularly in which extracellular ligand binding is a scFV derived from a CD33 monoclonal antibody, conferring specific immunity against CD33 positive cells. The engineered immune cells endowed with such CARs are particularly suited for treating lymphomas and leukemia.

1 . A CD33 specific chimeric antigen receptor (CAR) comprising:
 (a) an extracellular ligand binding domain comprising a heavy chain variable region (V H ) and a light chain variable region (V L ) from a monoclonal anti-CD33 antibody;   (b) a hinge selected from an FcRIIIα hinge, a CD8α hinge, and an IgG1 hinge;   (c) a CD8α transmembrane domain; and   (d) a cytoplasmic domain comprising a CD3-ζ signaling domain and a co-stimulatory domain from 4-1BB.   
     
     
         2 . The CAR of  claim 1 , comprising a CD8α hinge and a CD8α transmembrane domain. 
     
     
         3 . The CAR of  claim 1 , wherein said CD8α hinge has at least 80% sequence identity with SEQ ID NO. 4. 
     
     
         4 . The CAR of  claim 1 , comprising a polypeptide sequence having at least 80% sequence identity with SEQ ID NO. 27, SEQ ID NO. 28, SEQ ID NO. 29 or SEQ ID NO. 30. 
     
     
         5 . The CAR of  claim 1 , comprising an FcγRIIIα hinge and a CD8a transmembrane domain. 
     
     
         6 . The CAR of  claim 5 , wherein said FcγRIIIα hinge has at least 80% sequence identity with SEQ ID NO. 3. 
     
     
         7 . The CAR of  claim 1 , comprising a polypeptide sequence having at least 80% sequence identity with SEQ ID NO. 19, SEQ ID NO. 20, SEQ ID NO. 21 or SEQ ID NO. 22. 
     
     
         8 . The CAR of  claim 1 , comprising an IgG1 hinge and a CD8a transmembrane domain. 
     
     
         9 . The CAR of  claim 8 , wherein said IgG1 hinge has at least 80% sequence identity with SEQ ID NO. 5. 
     
     
         10 . The CAR of  claim 1 , comprising a polypeptide sequence having at least 80% sequence identity with SEQ ID NO. 35, SEQ ID NO. 36, SEQ ID NO. 37 or SEQ ID NO. 38. 
     
     
         11 . The CAR of  claim 1 , wherein the V H  region has at least 80% sequence identity with a polypeptide sequence selected from SEQ ID NO. 11, SEQ ID NO. 13, SEQ ID NO. 15 and SEQ ID NO. 17; and wherein the V L  region has at least 80% sequence identity with a polypeptide sequence selected from SEQ ID NO. 12, SEQ ID NO. 14, SEQ ID NO. 16 and SEQ ID NO. 18. 
     
     
         12 . The CAR of  claim 1 , wherein the co-stimulatory domain from 4-1BB has at least 80% sequence identity with SEQ ID NO. 8. 
     
     
         13 . The CAR of  claim 1 , wherein the CD3ζ signaling domain has at least 80% sequence identity with SEQ ID NO. 9. 
     
     
         14 . The CAR of  claim 1 , wherein said CD8a transmembrane domain has at least 80% sequence identity with SEQ ID NO. 6. 
     
     
         15 - 19 . (canceled) 
     
     
         20 . An engineered immune cell expressing at the cell surface membrane the CAR of  claim 1 . 
     
     
         21 . The engineered immune cell of  claim 20 , wherein the engineered immune cell is derived from at least one of an inflammatory T-lymphocyte, a cytotoxic T-lymphocyte, a regulatory T-lymphocyte, or a helper T-lymphocyte. 
     
     
         22 . The engineered immune cell of  claim 20 , wherein expression of T cell receptor (TCR) is suppressed. 
     
     
         23 . The engineered immune cell of  claim 20 , wherein expression of CD33 is suppressed. 
     
     
         24 . The engineered immune cell of  claim 20 , wherein said engineered immune cell is modified to be resistant to at least one of an immunosuppressive drug or a chemotherapy drug. 
     
     
         25 - 42 . (canceled) 
     
     
         43 . A composition comprising:
 a chimeric antigen receptor (CAR) comprising:
 (a) an extracellular ligand binding domain comprising a heavy chain variable region (V H ) and a light chain variable region (V L ) from a monoclonal anti-CD33 antibody; 
 (b) a hinge selected from an FcRIIIα hinge, a CD8a hinge, and an IgG1 hinge; 
 (c) a CD8α transmembrane domain; and 
 (d) a cytoplasmic domain comprising a CD3-ζ signaling domain and a co-stimulatory domain from 4-1BB.