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 nucleic acid comprising a polynucleotide, in order from 5′ to 3′, encoding
(a) a chimeric antigen receptor, comprising, in order from the amino terminus of the receptor, (i) an scFv antigen recognition moiety; (ii) a transmembrane region; and (iii) a CD3 zeta polypeptide;
(b) linker polypeptide, wherein the linker polypeptide is a cleavable 2A polypeptide that links the chimeric antigen receptor to the chimeric stimulating molecule; and
(c) a chimeric stimulating molecule, wherein:
the chimeric stimulating molecule comprises (i) a truncated MyD88 polypeptide lacking the TIR domain, and (ii) a CD40 cytoplasmic polypeptide region lacking the CD40 extracellular domain;
the chimeric stimulating molecule is constitutively active;
the chimeric stimulating molecule does not include a dimerization or multimerization molecule binding region; and
the polynucleotide is operably linked to a promoter.
2. The nucleic acid of claim 1 , wherein the chimeric stimulating molecule does not include a membrane-targeting region.
3. The nucleic acid of claim 1 , wherein the transmembrane region is a CD8 transmembrane region.
4. The nucleic acid of claim 3 , wherein the chimeric antigen receptor comprises a CD8 stalk region between the scFv antigen recognition moiety and the CD8 transmembrane region.
5. The nucleic acid of claim 1 , wherein the 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.
6. The nucleic acid of claim 1 , wherein the antigen recognition moiety binds to an antigen selected from the group consisting of PSCA, CD19, and Her2/Neu.
7. The nucleic acid of claim 1 , wherein the antigen recognition moiety binds to Her2/Neu.
8. The nucleic acid of claim 7 , wherein the antigen recognition moiety comprises the amino acid sequence of SEQ ID NO: 155 and the amino acid sequence of SEQ ID NO: 159.
9. The nucleic acid of claim 1 , wherein the antigen recognition moiety binds to CD19.
10. The nucleic acid of claim 9 , wherein the antigen recognition moiety comprises the amino acid sequence of SEQ ID NO: 131 and the amino acid sequence of SEQ ID NO: 135.
11. A modified T cell comprising the nucleic acid of claim 1 .
12. A modified T cell comprising the nucleic acid of claim 2 .
13. A modified T cell comprising the nucleic acid of claim 5 .
14. A modified cell comprising the nucleic acid of claim 9 .
15. The modified cell of claim 14 , wherein the modified cell is a T cell, tumor infiltrating lymphocyte, NK-T cell, TCR-expressing cell, or NK cell.
16. The modified cell of claim 14 , wherein the modified cell is a T cell.