Biologically relevant orthogonal cytokine/receptor pairs

Abstract

Engineered orthogonal cytokine receptor/ligand pairs, and methods of use thereof, are provided.

What is claimed is: 
     
         1 . An engineered human IL-2 polypeptide, which (i) has significantly reduced binding to native human CD122; and which (ii) comprises at least one amino acid substitution with an amino acid other than that of the native protein at residue T51, R81, or comprises amino acid substitution M23A; and (iii) comprises amino acid substitutions at each of E15, H16, L19, D20. 
     
     
         2 . The engineered human IL-2 polypeptide of  claim 1 , wherein the polypeptide comprises one or more amino acid substitutions selected from: [E15D, E15T, E15A, E15S], [H16N, H16Q], [L19V, L19I, L19A], [D20L, D20M], [Q22S, Q22T, Q22E, Q22K, Q22E], [M23A, M23W, M23H, M23Y, M23F, M23Q, M23Y], [G27K, G27S], [R81D, R81Y], [N88E, N88Q], [T51I]. 
     
     
         3 . The engineered human IL-2 polypeptide of  claim 1  or  claim 2 , wherein the polypeptide comprises amino acid substitutions E15S; H16Q; L19V; and D20L. 
     
     
         4 . The engineered human IL-2 polypeptide of any of  claims 1 - 3 , further comprising amino acid substitution Q22K. 
     
     
         5 . The engineered human IL-2 polypeptide of any of  claims 1 - 4 , comprising amino acid substitution T51I. 
     
     
         6 . The engineered human IL-2 polypeptide of any of  claims 1 - 5 , comprising amino acid substitution R81D or R81Y. 
     
     
         7 . The engineered human IL-2 polypeptide of  claim 1  comprising a set of amino acid substitutions [E15D, H16N, L19V, D20L, Q22T, M23A]. 
     
     
         8 . The engineered human IL-2 polypeptide of  claim 1  comprising a set of amino acid substitutions[E15D, H16N, L19V, D20L, Q22K, M23A]. 
     
     
         9 . The engineered human IL-2 polypeptide of  claim 1  comprising a set of amino acid substitutions [E15S, H16Q, L19V, D20L, M23Q, R81D, T51I]. 
     
     
         10 . The engineered human IL-2 polypeptide of  claim 1  comprising a set of amino acid substitutions [E15S, H16Q, L19V, D20L, M23Q, R81Y]. 
     
     
         11 . The engineered human IL-2 polypeptide of any of  claims 1 - 10 , wherein the polypeptide binds to and activates an orthogonal human CD122 protein. 
     
     
         12 . The engineered human IL-2 polypeptide of  claim 11 , wherein the orthogonal human CD122 protein is modified at one or more residues selected from R41, R42, Q70, K71, T73, T74, V75, S132, H133, Y134, F135, E136, Q214. 
     
     
         13 . The engineered human IL-2 polypeptide of  claim 12 , wherein the orthogonal human CD122 protein is modified at H133 and Y134. 
     
     
         14 . The engineered human IL-2 polypeptide of  claim 13 , wherein the orthogonal human CD122 protein comprises amino acid substitutions H133D and Y134F. 
     
     
         15 . A system for selective activation of a receptor in a cell, the system comprising:
 (a) an orthogonal human CD122 receptor comprising amino acid substitutions at H133 and Y134; and   (b) the engineered human IL-2 polypeptide of any of  claims 1 - 10 .   
     
     
         16 . The system of  claim 15 , wherein the orthogonal receptor is expressed by a mammalian cell. 
     
     
         17 . The system of  claim 16 , wherein the cell is an immune cell or a stem cell. 
     
     
         18 . The system of  claim 17 , wherein the immune cell is a T cell. 
     
     
         19 . The system of  claim 18 , wherein the T cell is a CAR-T cell. 
     
     
         20 . A pharmaceutical composition comprising the engineered human IL-2 polypeptide of any of  claims 1 - 10 ; and a pharmaceutically acceptable excipient. 
     
     
         21 . A nucleic acid encoding the engineered human IL-2 polypeptide of any of  claims 1 - 10 . 
     
     
         22 . An expression vector comprising the nucleic acid of  claim 21 . 
     
     
         23 . A cell genetically engineered to comprise the vector of  claim 22 . 
     
     
         24 . A method of treating an individual, the method comprising introducing an immune effector cell expressing an orthogonal human CD122 receptor comprising amino acid substitutions at H133 and Y134, and selectively activating the cell by contacting with an engineered human IL-2 polypeptide according to any of  claims 1 - 10 . 
     
     
         25 . The method of  claim 24 , wherein the immune effector cell is a T cell. 
     
     
         26 . The method of  claim 25 , wherein the T cell is a CAR-T cell. 
     
     
         27 . The method of any of  claims 24 - 26 , wherein the individual is treated for cancer. 
     
     
         28 . The method of any of  claims 24 - 26 , wherein the individual is treated for autoimmune disease. 
     
     
         29 . The method of any of  claims 24 - 26 , wherein the individual is treated for infection. 
     
     
         30 . A kit comprising the system of  claim 15 .