Chimeric receptors and methods of use thereof

Abstract

Antigen binding molecules, chimeric receptors, and engineered immune cells are disclosed in accordance with the invention. The invention further relates to vectors, compositions, and methods of treatment and/or detection using the antigen binding molecules and engineered immune cells.

1 - 80 . (canceled) 
     
     
         81 . A method for preparing an immune cell, the method comprising introducing to the immune cell a polynucleotide encoding a chimeric antigen receptor (CAR) that binds to C-type lectin-like-1 (“CLL-1”), wherein the CAR comprises a single chain Fv (scFv) that binds CLL-1, a transmembrane domain, and an intracellular activating domain that is a signaling domain of CD3 zeta, and wherein the scFv comprises a heavy chain variable region (VH) comprising VH complementarity determining regions (“CDRs”) 1, VH CDR2, and VH CDR3, and a light chain variable region (VL) comprising VL CDR1, VL CDR2, and VL CDR3, wherein:
 (a) the VH CDR1 comprises the amino acid sequence of SEQ ID NO: 17 or a variant derived therefrom with an amino acid substitution,
 the VH CDR2 comprises the amino acid sequence of SEQ ID NO: 18 or a variant derived therefrom with an amino acid substitution, 
 the VH CDR3 comprises the amino acid sequence of SEQ ID NO: 19 or a variant derived therefrom with an amino acid substitution, 
 the VL CDR1 comprises the amino acid sequence of SEQ ID NO: 22 or a variant derived therefrom with an amino acid substitution, 
 the VL CDR2 comprises the amino acid sequence of SEQ ID NO: 23 or a variant derived therefrom with an amino acid substitution, and 
 the VL CDR3 comprises the amino acid sequence of SEQ ID NO: 24 or a variant derived therefrom with an amino acid substitution; 
 
 (b) the VH CDR1 comprises the amino acid sequence of SEQ ID NO: 51 or a variant derived therefrom with an amino acid substitution,
 the VH CDR2 comprises the amino acid sequence of SEQ ID NO: 52 or a variant derived therefrom with an amino acid substitution, 
 the VH CDR3 comprises the amino acid sequence of SEQ ID NO: 53 or a variant derived therefrom with an amino acid substitution, 
 the VL CDR1 comprises the amino acid sequence of SEQ ID NO: 56 or a variant derived therefrom with an amino acid substitution, 
 the VL CDR2 comprises the amino acid sequence of SEQ ID NO: 57 or a variant derived therefrom with an amino acid substitution, and 
 the VL CDR3 comprises the amino acid sequence of SEQ ID NO: 58 or a variant derived therefrom with an amino acid substitution; 
 
 (c) the VH CDR1 comprises the amino acid sequence of SEQ ID NO: 73 or a variant derived therefrom with an amino acid substitution,
 the VH CDR2 comprises the amino acid sequence of SEQ ID NO: 74 or a variant derived therefrom with an amino acid substitution, 
 the VH CDR3 comprises the amino acid sequence of SEQ ID NO: 75 or a variant derived therefrom with an amino acid substitution, 
 the VL CDR1 comprises the amino acid sequence of SEQ ID NO: 78 or a variant derived therefrom with an amino acid substitution, 
 the VL CDR2 comprises the amino acid sequence of SEQ ID NO: 79 or a variant derived therefrom with an amino acid substitution, and 
 the VL CDR3 comprises the amino acid sequence of SEQ ID NO: 80 or a variant derived therefrom with an amino acid substitution; or 
 
 (d) the VH CDR1 comprises the amino acid sequence of SEQ ID NO: 95 or a variant derived therefrom with an amino acid substitution,
 the VH CDR2 comprises the amino acid sequence of SEQ ID NO: 96 or a variant derived therefrom with an amino acid substitution, 
 the VH CDR3 comprises the amino acid sequence of SEQ ID NO: 97 or a variant derived therefrom with an amino acid substitution, 
 the VL CDR1 comprises the amino acid sequence of SEQ ID NO: 100 or a variant derived therefrom with an amino acid substitution, 
 the VL CDR2 comprises the amino acid sequence of SEQ ID NO: 101 or a variant derived therefrom with an amino acid substitution, and 
 the VL CDR3 comprises the amino acid sequence of SEQ ID NO: 102 or a variant derived therefrom with an amino acid substitution. 
 
 
     
     
         82 . The method of  claim 81 , wherein:
 (a) the VH comprises the amino acid sequence of SEQ ID NO: 16 or a sequence having at least 95% sequence identity to SEQ ID NO:16, and the VL comprises the amino acid sequence of SEQ ID NO: 21 or a sequence having at least 95% sequence identity to SEQ ID NO:21,   (b) the VH comprises the amino acid sequence of SEQ ID NO: 50 or a sequence having at least 95% sequence identity to SEQ ID NO:50, and the VL comprises the amino acid sequence of SEQ ID NO: 55 or a sequence having at least 95% sequence identity to SEQ ID NO:55,   (c) the VH comprises the amino acid sequence of SEQ ID NO: 72 or a sequence having at least 95% sequence identity to SEQ ID NO:72, and the VL comprises the amino acid sequence of SEQ ID NO: 77 or a sequence having at least 95% sequence identity to SEQ ID NO:77, or   (d) the VH comprises the amino acid sequence of SEQ ID NO: 94 or a sequence having at least 95% sequence identity to SEQ ID NO:94, and the VL comprises the amino acid sequence of SEQ ID NO: 99 or a sequence having at least 95% sequence identity to SEQ ID NO:99.   
     
     
         83 . The method of  claim 81 , wherein the signaling domain of CD3 zeta comprises the amino acid sequence of SEQ ID NO: 10 or a sequence having at least 95% sequence identity to SEQ ID NO:10. 
     
     
         84 . The method of  claim 81 , wherein the CAR further comprises a costimulatory domain signaling region. 
     
     
         85 . The method of  claim 84 , wherein the costimulatory domain signaling region is a signaling region of CD28, OX-40, 4-1BB, CD27, or ICOS. 
     
     
         86 . The method of  claim 84 , wherein the costimulatory domain signaling region comprises the amino acid sequence of SEQ ID NO: 8 or a sequence having at least 95% sequence identity to SEQ ID NO:8. 
     
     
         87 . The method of  claim 86 , wherein the CAR comprises the amino acid sequence of SEQ ID NO: 2 or a sequence having at least 95% sequence identity to SEQ ID NO:2. 
     
     
         88 . The method of  claim 81 , wherein the transmembrane domain is a CD28 or a CD8 transmembrane domain. 
     
     
         89 . The method of  claim 88 , wherein the transmembrane domain comprises the amino acid sequence of SEQ ID NO: 6 or a sequence having at least 95% sequence identity to SEQ ID NO:6. 
     
     
         90 . The method of  claim 88 , wherein the CAR comprises the amino acid sequence of SEQ ID NO: 2 or 14, or a sequence having at least 95% sequence identity to SEQ ID NO:2 or 14. 
     
     
         91 . The method of  claim 81 , wherein scFv further comprises a linker comprising the amino acid sequence of SEQ ID NO: 130 or 132, or a sequence having at least 95% sequence identity to SEQ ID NO:130 or 132. 
     
     
         92 . The method of  claim 81 , wherein the CAR comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 60, 62, 64, 66, 68, 70, 82, 84, 86, 88, 90, 92, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, and 126, or a sequence having at least 90% sequence identity to SEQ ID NO: 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 60, 62, 64, 66, 68, 70, 82, 84, 86, 88, 90, 92, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, or 126. 
     
     
         93 . The method of  claim 81 , wherein the polynucleotide comprises a nucleic sequence selected from the group consisting of SEQ ID NO: 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 59, 61, 63, 65, 67, 69, 81, 83, 85, 87, 89, 91, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, and 125 or a sequence having at least 90% sequence identity to SEQ ID NO: 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46, 48, 60, 62, 64, 66, 68, 70, 82, 84, 86, 88, 90, 92, 104, 106, 108, 110, 112, 114, 116, 118, 120, 122, 124, or 126. 
     
     
         94 . The method of  claim 81 , wherein the polynucleotide is provided in a vector. 
     
     
         95 . The method of  claim 94 , wherein the vector a retroviral vector, a DNA vector, a plasmid, a RNA vector, an adenoviral vector, an adenovirus associated vector, or a lentiviral vector. 
     
     
         96 . The method of  claim 81 , wherein the immune cell is a T cell, a tumor infiltrating lymphocyte (TIL), an NK cell, or an NK-T cell. 
     
     
         97 . The method of  claim 96 , wherein the immune cell is an autologous cell. 
     
     
         98 . The method of  claim 96 , wherein the immune cell is an allogeneic cell. 
     
     
         99 . The method of  claim 81 , wherein the introducing is by transfection or transduction. 
     
     
         100 . The method of  claim 99 , wherein the transfection or transduction is in vivo, in vitro, or ex vivo.