Chimeric molecules providing targeted costimulation for adoptive cell therapy

Abstract

The present invention relates to a chimeric molecule useful in adoptive cell therapy (ACT), and cells comprising the same. The chimeric molecule can act as a modulator of cellular activity enhancing responses when an endogenous T-cell receptor (TCR) is engaged with its cognate antigen. The present invention also provides proteins, nucleic acids encoding the chimeric molecule and therapeutic uses thereof.

What is claimed is: 
     
         1 . An engineered protein that has at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 100%, or any integer that is between 80 and 100%, identity to SEQ ID NO: 166, and wherein the sequence is not SEQ ID NO: 123. 
     
     
         2 . An engineered protein that has at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 100%, or any integer that is between 80 and 100%, identity to SEQ ID NO: 167, and wherein the sequence is not SEQ ID NO: 123. 
     
     
         3 . The engineered protein of any one of  claim 1  or  2 , further comprising a binding domain, CD28 domain, and CD40 domain. 
     
     
         4 . The engineered protein of any one of  claims 2 - 3 , further comprising a signal peptide sequence. 
     
     
         5 . The engineered protein of  claim 4 , wherein the signal peptide sequence has at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 100%, or any integer that is between 80 and 100%, identity to the amino acid sequence of SEQ ID NO: 157. 
     
     
         6 . The engineered protein of any one of  claims 3 - 5 , wherein the binding domain comprises a VL sequence, a VH sequence, and an at least one linker. 
     
     
         7 . The engineered protein of  claim 6 , wherein the at least one linker has at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 100%, or any integer that is between 80 and 100%, identity to the amino acid sequence of SEQ ID NO: 159 or 161. 
     
     
         8 . The engineered protein of any one of  claims 6 - 7 , wherein the VL sequence has at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 100%, or any integer that is between 80 and 100%, identity to the amino acid sequence of SEQ ID NO: 158. 
     
     
         9 . The engineered protein of any one of  claims 6 - 8 , wherein the VH sequence has at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 100%, or any integer that is between 80 and 100%, identity to the amino acid sequence of SEQ ID NO: 160. 
     
     
         10 . The engineered protein of any one of  claims 3 - 9 , wherein the CD40 domain has at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 100%, or any integer that is between 80 and 100%, identity to the amino acid sequence of SEQ ID NO: 165. 
     
     
         11 . The engineered protein of any one of  claims 3 - 10 , wherein the CD28 domain comprises a CD28 transmembrane domain. 
     
     
         12 . The engineered protein of  claim 11 , wherein the CD28 transmembrane domain has at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 100%, or any integer that is between 80 and 100%, identity to SEQ ID NO: 163. 
     
     
         13 . The engineered protein of any one of  claims 3 - 12 , wherein the CD28 domain comprises a CD28 extracellular domain. 
     
     
         14 . The engineered protein of  claim 13 , wherein the CD28 extracellular domain has at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 100%, or any integer that is between 80 and 100%, identity to SEQ ID NO: 162. 
     
     
         15 . The engineered protein of any one of  claims 3 - 14 , wherein the CD28 domain comprises a CD28 intracellular domain. 
     
     
         16 . The engineered protein of  claim 15 , wherein the CD28 intracellular domain has at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 100%, or any integer that is between 80 and 100%, identity to SEQ ID NO: 164. 
     
     
         17 . The engineered protein of any one of  claims 1 - 16 , wherein the protein further comprises 1, 2, 3, 4, 5, or all 6 CDR sequence(s) selected from the group consisting of: QASQSLSNLLA (SEQ ID NO: 168), GASNLES (SEQ ID NO: 169), QGGHYSGL (SEQ ID NO: 170), TNDMN (SEQ ID NO: 171), VIYSDDTPDYATWAKG (SEQ ID NO: 172), and/or GHYDASVYAYALNI (SEQ ID NO: 173). 
     
     
         18 . An engineered protein comprising an amino acid sequence that is at least 80% identical to the amino acid sequence of SEQ ID NO: 166 or 167, wherein the amino acid sequence does not include at least one of: QKLISEEDLE (SEQ ID NO: 174) or 
       
         
           
                 
               
                   (SEQ ID NO: 175) 
                 
                   LVKQSPMLVAYDNAVNLSCKYSYNLFSREFRASLHKGLDSAVEVCVVYGN 
                 
                     
                 
                   YSQQLQVYSKTGFNCDGKLGNESVTFYLQNLYVNQTDIYFCKI. 
                 
             
                
                
                
                
               
            
           
         
       
     
     
         19 . A CoStAR comprising:
 (a) an optional signal peptide;   (b) a binding domain, wherein the binding domain binds to an anti-pembrolizumab antibody or binding fragment thereof;   (c) a CD28 domain;   (d) a CD40 domain;   
       wherein a) is optionally linked to b), wherein b) is linked to c), wherein c) is linked to d), and 
       wherein the CoStAR comprises an amino acid sequence that:
 i) lacks at least one of: QKLISEEDLE (SEQ ID NO: 174) or 
 
       
         
           
                 
               
                   (SEQ ID NO: 175) 
                 
                   LVKQSPMLVAYDNAVNLSCKYSYNLFSREFRASLHKGLDSAVEVCVVYGN 
                 
                     
                 
                   YSQQLQVYSKTGFNCDGKLGNESVTFYLQNLYVNQTDIYFCKI; 
                 
             
                
                
                
                
               
            
           
         
         ii) has an amino acid sequence that is greater than 95% identical to SEQ ID NO: 166 or 167; 
         iii) has an amino acid sequence that is greater than 80% identical to SEQ ID NO: 166 or 167 and is not SEQ ID NO: 123; or 
         iv) any combination of i-iv. 
       
     
     
         20 . A fusion protein comprising:
 (a) a means for binding to an antibody that binds to pembrolizumab;   (b) a CD28 domain;   (c) a CD40 domain;   wherein a) is linked to b), wherein b) is linked to c), and wherein the fusion protein comprises an amino acid sequence that:   i) lacks at least one of: QKLISEEDLE (SEQ ID NO: 174) or LVKQSPMLVAYDNAVNLSCKYSYNLFSREFRASLHKGLDSAVEVCVVYGNYSQQLQV YSKTGFNCDGKLGNESVTFYLQNLYVNQTDIYFCKI (SEQ ID NO: 175);   ii) has an amino acid sequence that is greater than 95% identical to SEQ ID NO: 166 or 167;   iii) has an amino acid sequence that is greater than 80% identical to SEQ ID NO: 166 or 167 and is not SEQ ID NO: 123; or   iv) any combination of i-iv.   
     
     
         21 . The CoStAR or fusion protein of  claim 19  or  20 , wherein the binding domain or the means for binding to an antibody that binds to pembrolizumab comprises: 1, 2, 3, 4, 5, or all 6 CDR sequence(s) selected from the group consisting of: QASQSLSNLLA (SEQ ID NO: 168), GASNLES (SEQ ID NO: 169), QGGHYSGL (SEQ ID NO: 170), TNDMN (SEQ ID NO: 171), VIYSDDTPDYATWAKG (SEQ ID NO: 172), and/or GHYDASVYAYALNI (SEQ ID NO: 173). 
     
     
         22 . A fusion protein comprising the amino acid sequence of SEQ ID NO: 166. 
     
     
         23 . A fusion protein comprising the amino acid sequence of SEQ ID NO: 167. 
     
     
         24 . A nucleic acid which encodes the protein of any one of the preceding claims. 
     
     
         25 . A vector which comprises the nucleic acid of any one of the preceding claims. 
     
     
         26 . A cell which expresses the protein of any one of the preceding claims. 
     
     
         27 . A cell which expresses at least two proteins of any one of the preceding claims. 
     
     
         28 . A method of making the cell of any one of  claim 26  or  27 , which comprises the step of transducing or transfecting a cell with a vector of  claim 25 . 
     
     
         29 . A method for preparing a population of cells that express a protein of any one of  claims 1  to  23 , comprising detecting expression of the protein on the surface of cells transfected or transduced with a vector according to  claim 25  and selecting cells which are identified as expressing the protein. 
     
     
         30 . A cell population which is enriched for cell expression a protein of any one of  claims 1  to  23 . 
     
     
         31 . A method for treating a disease in a subject in need thereof, which comprises the step of administering the cell of any one of  claims 26 - 27  or the cell population of  claim 30  to the subject.