Heterodimeric proteins for modulating gamma delta t cells

Abstract

The present technology relates, inter alia, to compositions and methods, including heterodimeric proteins that find use in the treatment of disease, such as immunotherapies for cancer and autoimmunity.

1 .- 74 . (canceled) 
     
     
         75 . A heterodimeric protein comprising an alpha chain and a beta chain wherein the alpha chain and the beta chain each independently comprise:
 (a) a first domain comprising one or more butyrophilin family proteins selected from BTN3A1, BTN3A2, BTN2A1, BTNL3, and BTNL8, or fragments thereof comprising the extracellular domains of the butyrophilin family proteins;   (b) a second domain comprising a targeting domain selected from an antibody (scFv), or a single chain antibody, wherein the targeting domain is capable of binding an antigen on the surface of a cancer cell; and   (c) a linker that adjoins the first and second domain, wherein:
 the linker of one of the alpha and beta chains comprises a first charge polarized core domain comprising:
 a peptide comprising positively charged amino acid residues at the amino terminus of the first charge polarized core domain, and 
 a peptide comprising negatively charged amino acid residues at the carboxy terminus of the first charge polarized core domain; and 
 
 the linker of other of the alpha and beta chains comprises a second charge polarized core domain comprising:
 a peptide comprising negatively charged amino acid residues at the amino terminus of the second charge polarized core domain, and 
 a peptide comprising positively charged amino acid residues at the carboxy terminus of the second charge polarized core domain. 
 
   
     
     
         76 . The heterodimeric protein of  claim 75 , wherein the alpha chain and the beta chain self-associate to form a heterodimer. 
     
     
         77 . The heterodimeric protein of  claim 75 , wherein the targeting domain comprises an scFv. 
     
     
         78 . The heterodimeric protein of  claim 75 , wherein the first domain of alpha and beta chains comprises:
 (a) human BTNL3, and human BTNL8;   (b) human BTN2A1 and human BTN3A1; or   (b) human BTN3A1 and human BTN3A2.   
     
     
         79 . The heterodimeric protein of  claim 81 , wherein the first domain is capable of binding a yb (gamma delta) T cell receptor. 
     
     
         80 . The heterodimeric protein of  claim 75 , wherein the first domain comprises a polypeptide having an amino acid sequence that has at least 95% identity with an amino acid sequence selected from SEQ ID NOs: 49-52 and 102. 
     
     
         81 . The heterodimeric protein of  claim 75 , wherein the linker comprises:
 (a) the first charge polarized core domain adjoined to the first domain of alpha chain at the carboxy terminus of the first domain of alpha chain, and   (b) the second charge polarized core domain adjoined to the first domain of beta chain at the carboxy terminus the first domain of beta chain.   
     
     
         82 . The heterodimeric protein of  claim 75 , wherein the linker comprises:
 (a) the first charge polarized core domain adjoined to the targeting domain of alpha chain at the amino terminus of the targeting domain of alpha chain, and   (b) the second charge polarized core domain adjoined to the targeting domain of beta chain at the amino terminus of the targeting domain of beta chain.   
     
     
         83 . The heterodimeric protein of  claim 75 , wherein the peptide comprising negatively charged amino acid residues comprises the sequence DEGGED (SEQ ID NO: 13) or GSGSDEGGEDGS (SEQ ID NO: 14). 
     
     
         84 . The heterodimeric protein of  claim 75 , wherein the peptide comprising positively charged amino acid residues comprises the sequence RKGGKR (SEQ ID NO: 11) or GSGSRKGGKRGS (SEQ ID NO: 12). 
     
     
         85 . The heterodimeric protein of  claim 75 , wherein:
 the peptide comprising positively charged amino acid residues comprises the sequence RKGGKR (SEQ ID NO: 11) or GSGSRKGGKRGS (SEQ ID NO: 12); and   the peptide comprising negatively charged amino acid residues comprises the sequence DEGGED (SEQ ID NO: 13) or GSGSDEGGEDGS (SEQ ID NO: 14).   
     
     
         86 . The linker of  claim 85 , wherein the linker forms a heterodimer through electrostatic interactions between positively charged amino acid residues and negatively charged amino acid residues on the first and second charge polarized core domains. 
     
     
         87 . The heterodimeric protein of  claim 75 , wherein the linker further comprises the hinge-CH2-CH3 Fc domain derived from IgG1 or the hinge-CH2-CH3 Fc domain derived from IgG4. 
     
     
         88 . The heterodimeric protein of  claim 85 , wherein the linker further comprises the hinge-CH2-CH3 Fc domain derived from IgG1 or the hinge-CH2-CH3 Fc domain derived from IgG4. 
     
     
         89 . The heterodimeric protein of  claim 75 , wherein the first domain and/or the heterodimeric protein modulates or is capable of modulating a gamma delta T cell. 
     
     
         90 . The heterodimeric protein of  claim 89 , wherein the gamma delta T cell is selected from a cell expressing Vγ4, Vγ9δ2, or Vγ7δ4. 
     
     
         91 . The heterodimeric protein of  claim 89 , wherein the modulation of a gamma delta T cell is activation of a gamma delta T cell. 
     
     
         92 . The heterodimeric protein of  claim 75 , wherein the heterodimeric protein is capable of forming a synapse between a gamma delta T cell and a tumor cell and/or the heterodimeric protein is capable of contemporaneous activation and targeting of gamma delta T cells to tumor cells. 
     
     
         93 . A pharmaceutical composition, comprising the heterodimeric protein of  claim 75 , and a pharmacologically acceptable excipient.