Costimulation of chimeric antigen receptors by myd88 and cd40 polypeptides

Abstract

The technology relates generally to the field of immunology and relates in part to methods for activating T cells and other cells resulting in an immune response against a target antigen. The technology also relates to costimulation of therapeutic cells that express chimeric antigen receptors that recognize target antigens using chimeric MyD88- and CD40-derived polypeptides. The technology further relates in part to therapeutic cells that express chimeric antigen receptors, wherein the chimeric antigen receptors have an endodomain that includes MyD88- and CD40-derived polypeptides, and methods for treating patients using the modified therapeutic cells.

1 . A nucleic acid comprising a promoter operably linked to a polynucleotide encoding a chimeric stimulating molecule, wherein the chimeric stimulating molecule comprises
 (i) a MyD88 polypeptide or a truncated MyD88 polypeptide lacking the TIR domain;   (ii) a CD40 cytoplasmic polypeptide region lacking the CD40 extracellular domain; and   (iii) a membrane targeting region, wherein the membrane targeting region is optionally selected from the group consisting of a myristoylation region, palmitoylation region, prenylation region, and transmembrane sequences of receptors.   
     
     
         2 . A nucleic acid comprising a promoter operably linked to a polynucleotide encoding a cytoplasmic chimeric stimulating molecule, wherein the cytoplasmic chimeric stimulating molecule comprises
 (i) a MyD88 polypeptide or a truncated MyD88 polypeptide lacking the TIR domain; and   (ii) a CD40 cytoplasmic polypeptide region lacking the CD40 extracellular domain.   
     
     
         3 . A nucleic acid comprising a promoter operably linked to a polynucleotide encoding a chimeric antigen receptor, wherein the chimeric antigen receptor comprises
 (i) a transmembrane region;   (ii) a MyD88 polypeptide or a truncated MyD88 polypeptide lacking a TIR domain;   (iii) a CD40 cytoplasmic polypeptide region lacking a CD40 extracellular domain; (iv) a T cell activation molecule; and   (v) an antigen recognition moiety,   wherein the chimeric antigen receptor optionally comprises (i) a transmembrane region; (ii) a T cell activation molecule; and (iii) an antigen recognition moiety, and   wherein the nucleic acid optionally further comprises a polynucleotide encoding a chimeric Caspase-9 polypeptide comprising a multimeric ligand binding region and a Caspase-9 polypeptide.   
     
     
         4 . The nucleic acid of  claim 1 , further comprising:
 (a) a polynucleotide encoding a chimeric antigen receptor; or   (b) a polynucleotide encoding a T cell receptor or a T cell receptor based chimeric antigen receptor.   
     
     
         5 . (canceled) 
     
     
         6 . The nucleic acid of  claim 1 , further comprising a second polynucleotide encoding a chimeric Caspase-9 polypeptide comprising a multimeric ligand binding region and a Caspase-9 polypeptide. 
     
     
         7 . The nucleic acid of  claim 1 , further comprising
 a) a second polynucleotide encoding a chimeric antigen receptor, a T cell receptor, or a T cell receptor based chimeric antigen receptor; and   b) a third polynucleotide encoding a chimeric Caspase-9 polypeptide comprising a multimeric ligand binding region and a Caspase-9 polypeptide, wherein the multimeric ligand binding region is optionally an FKBP12 region, and   wherein one promoter is optionally operably linked to the chimeric stimulating molecule-encoding polynucleotide, and the second, and the third polynucleotides.   
     
     
         8 . The nucleic acid of  claim 6 , wherein one promoter is operably linked to both the chimeric stimulating molecule-encoding polynucleotide and the second polynucleotide, and
 wherein the nucleic acid optionally further comprises a linker polynucleotide encoding a linker polypeptide between the polynucleotides, wherein the linker polypeptide separates the translation products of the during or after translation.   
     
     
         9 .- 12 . (canceled) 
     
     
         13 . The nucleic acid of  claim 4 , wherein the chimeric antigen receptor comprises (i) a transmembrane region; (ii) a T cell activation molecule; and (iii) an antigen recognition moiety. 
     
     
         14 .- 15 . (canceled) 
     
     
         16 . A chimeric stimulating molecule polypeptide encoded by a nucleic acid of  claim 1 . 
     
     
         17 . A chimeric stimulating molecule polypeptide encoded by a nucleic acid of  claim 2 . 
     
     
         18 . A chimeric antigen receptor encoded by a nucleic acid of  claim 3 . 
     
     
         19 . A modified cell transfected or transduced with a nucleic acid of  claim 1 , wherein the modified cell optionally further comprises a nucleic acid comprising a polynucleotide encoding a chimeric antigen receptor, and
 wherein the modified cells are optionally transfected or transduced ex vivo.   
     
     
         20 . A modified cell transfected or transduced with a nucleic acid of  claim 13 ,
 wherein the modified cell is optionally a T cell, tumor infiltrating lymphocyte, NK-T cell, TCR-expressing cell, or NK cell, and   wherein the antigen recognition moiety optionally binds to an antigen selected from the group consisting of PSMA, PSCA, MUC1, CD19, ROR1, Mesothelin, GD2, CD123, MUC16, Her2/Neu, CD20, CD30, PRAME, NY-ESO-1, and EGFRvIII.   
     
     
         21 .- 22 . (canceled) 
     
     
         23 . A modified cell transduced or transfected with a nucleic acid of  claim 3 . 
     
     
         24 . (canceled) 
     
     
         25 . A method for stimulating a T cell-mediated immune response in a subject, comprising administering an effective amount of modified cells of  claim 20 ,
 wherein the T cell-mediated immune response is optionally directed against a target cell, and wherein the target cell is optionally a tumor cell.   
     
     
         26 .- 27 . (canceled) 
     
     
         28 . The method of  claim 25 , comprising measuring the number or concentration of target cells in a first sample obtained from the subject before administering the modified cell, measuring the number concentration of target cells in a second sample obtained from the subject after administration of the modified cell, and determining an increase or decrease of the number or concentration of target cells in the second sample compared to the number or concentration of target cells in the first sample. 
     
     
         29 . A method for treating a subject having a disease or condition associated with an elevated expression of a target antigen, comprising administering to the subject an effective amount of the modified cell of  claim 20 ,
 wherein the modified cell optionally comprises a chimeric Caspase-9 polypeptide comprising a multimeric ligand binding region and a Caspase-9 polypeptide, and the method optionally further comprises administering a multimeric ligand that binds to the multimeric ligand binding region to the subject following administration of the modified cells to the subject, and   wherein optionally after administration of the multimeric ligand, the number of modified cells comprising the chimeric Caspase-9 polypeptide is reduced.   
     
     
         30 .- 31 . (canceled) 
     
     
         32 . The method of  claim 29 , wherein the modified cells are transfected or transduced in vivo. 
     
     
         33 . (canceled) 
     
     
         34 . The nucleic acid of  claim 2 , further comprising a polynucleotide encoding a chimeric antigen receptor. 
     
     
         35 . A modified cell comprising a nucleic acid of  claim 34 .