Costimulation of chimeric antigen receptors by MYD88 and CD40 polypeptides
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 modified cell comprising a nucleic acid comprising a first 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 chimeric stimulating molecule does not include a multimeric ligand binding region, and
wherein the chimeric stimulating molecule is constitutively active.
2. The modified cell of claim 1 , wherein the membrane targeting region is selected from the group consisting of a myristoylation region, a palmitoylation region, a prenylation region, and a transmembrane sequences of receptors.
3. The modified cell of claim 1 , wherein the first polynucleotide is operably linked to a promoter.
4. The modified cell of claim 1 , wherein the nucleic acid further comprises a second polynucleotide encoding a chimeric antigen receptor, comprising:
(i) an scFv antigen recognition moiety;
(ii) a transmembrane region; and
(iii) a CD3 zeta polypeptide.
5. The modified cell of claim 4 , wherein the chimeric antigen receptor comprises a CD8 stalk region between the scFv antigen recognition moiety and the CD8 transmembrane region.
6. The modified cell of claim 4 , wherein the scFv antigen recognition moiety 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.
7. The modified cell of claim 4 , wherein the scFv antigen recognition moiety binds to an antigen selected from the group consisting of PSCA, CD19, and Her2/Neu.
8. The modified cell of claim 7 , wherein the scFv antigen recognition moiety binds to Her2/Neu.
9. The modified cell of claim 4 , wherein the scFv antigen recognition moiety comprises the amino acid sequence of SEQ ID NO: 155 and the amino acid sequence of SEQ ID NO: 159.
10. The modified cell of claim 7 , wherein the scFv antigen recognition moiety binds to CD19.
11. The modified cell of claim 4 , wherein the scFv antigen recognition moiety comprises the amino acid sequence of SEQ ID NO: 131 and the amino acid sequence of SEQ ID NO: 135.
12. The modified cell of claim 1 , wherein the modified cell is a T cell, a tumor infiltrating lymphocyte, a NK-T cell, a TCR-expressing cell, or a NK cell.
13. The modified cell of claim 1 , wherein the modified cell is a T cell.
14. The modified cell of claim 1 , wherein the modified cell is a NK cell.
15. The modified cell of claim 2 , wherein the membrane targeting region is a myristoylation region.
16. The modified cell of claim 1 , wherein the truncated MyD88 polypeptide has the amino acid sequence of SEQ ID NO: 147.
17. The modified cell of claim 1 , wherein the MyD88 polypeptide has the amino acid sequence of SEQ ID NO: 282.
18. The modified cell of claim 1 , wherein the cytoplasmic CD40 polypeptide has the amino acid sequence of SEQ ID NO: 149, or a functional fragment thereof.
19. The modified cell of claim 1 , wherein the nucleic acid further comprises a third polynucleotide encoding a chimeric Caspase-9 polypeptide comprising a multimeric ligand binding region and a Caspase-9 polypeptide.
20. The modified cell of claim 19 , wherein the Caspase-9 polypeptide comprises the amino acid sequence of SEQ ID NO: 153.
21. The modified cell of claim 19 , wherein the multimeric ligand binding region is an FKBP12 region.
22. The modified cell of claim 21 , wherein the FKBP12 region is an FKBP12v36 region.