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EP-3189148-B1 - COSTIMULATION OF CHIMERIC ANTIGEN RECEPTORS BY MYD88 AND CD40 POLYPEPTIDES

EP3189148B1EP 3189148 B1EP3189148 B1EP 3189148B1EP-3189148-B1

Inventors

  • SPENCER, DAVID
  • FOSTER, Aaron, Edward
  • SLAWIN, KEVIN

Dates

Publication Date
20260506
Application Date
20150901

Claims (20)

  1. A nucleic acid comprising a promoter operably linked to a polynucleotide encoding a cytoplasmic chimeric stimulating protein, wherein the cytoplasmic chimeric stimulating protein 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; with the proviso that the cytoplasmic chimeric stimulating protein does not include a multimerizing or dimerizing ligand binding site.
  2. The nucleic acid of claim 1, wherein the cytoplasmic chimeric stimulating protein is constitutively active.
  3. The nucleic acid of claims 1 or 2, wherein the nucleic acid further comprises a second polynucleotide encoding a chimeric antigen receptor, or a T cell receptor.
  4. The nucleic acid of claim 3, wherein the second polynucleotide encodes a chimeric antigen receptor comprising a transmembrane region, a T cell activation molecule, and an antigen recognition moiety.
  5. The nucleic acid of claim 4, wherein the chimeric antigen receptor is encoded 5' of the cytoplasmic chimeric stimulating protein.
  6. The nucleic acid of any one of clams 3 to 5, wherein one promoter is operably linked to both the first and second polynucleotides.
  7. The nucleic acid of any one of claims 3 to 5, comprising a polynucleotide encoding a 2A polypeptide linker between the first polynucleotide and the second polynucleotide.
  8. A nucleic acid comprising a promoter operably linked to a polynucleotide encoding a chimeric antigen receptor polypeptide, wherein the chimeric antigen receptor polypeptide 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.
  9. The nucleic acid of any one of claims 1 to 8, wherein the nucleic acid comprises a polynucleotide that encodes a chimeric Caspase-9 polypeptide comprising a multimeric ligand binding region and a Caspase-9 polypeptide.
  10. A modified cell comprising a nucleic acid of claims 1 or 2.
  11. A modified cell comprising a nucleic acid of any one of claims 3 to 9.
  12. A modified cell comprising a nucleic acid of any one of claims 1 to 2, wherein the modified cell comprises a nucleic acid comprising a polynucleotide encoding a chimeric antigen receptor, or a T cell receptor.
  13. A modified cell comprising a nucleic acid of any one of claims 1 to 8, wherein the modified cell comprises a polynucleotide encoding a chimeric Caspase-9 polypeptide, wherein the chimeric Caspase-9 polypeptide comprises a multimeric ligand binding region and a Caspase-9 polypeptide.
  14. The nucleic acid of any one of claims 4 to 8, or a modified cell of claim 11 or 12, wherein the chimeric antigen receptor comprises an antigen recognition moiety that 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.
  15. The modified cell of any one of claims 10 to 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 15, wherein the cell is a T cell.
  17. A chimeric antigen receptor polypeptide encoded by a nucleic acid of claim 8.
  18. A modified cell of any one of claims 10 to 16 for use in stimulating a T cell-mediated immune response in a subject.
  19. A modified cell of any one of claims 10 to 16 for use in providing anti-tumor immunity to a subject.
  20. A modified cell of any one of claims 10 to 16 for use in treating a subject having a disease or condition associated with an elevated expression of a target antigen.

Description

Field The present invention is defined by the claims and the present disclosure 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 present disclosure also relates to costimulation of therapeutic cells that express chimeric antigen receptors that recognize target antigens using chimeric MyD88- and CD40- derived polypeptides. The present disclosure 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. Background T cell activation is an important step in the protective immunity against pathogenic microorganisms (e.g., viruses, bacteria, and parasites), foreign proteins, and harmful chemicals in the environment, and also as immunity against cancer and other hyperproliferative diseases. T cells express receptors on their surfaces (i.e., T cell receptors) that recognize antigens presented on the surface of cells. During a normal immune response, binding of these antigens to the T cell receptor, in the context of MHC antigen presentation, initiates intracellular changes leading to T cell activation. Chimeric antigen receptors (CARs) are artificial receptors designed to convey antigen specificity to T cells without the requirement for MHC antigen presentation. They include an antigen-specific component, a transmembrane component, and an intracellular component selected to activate the T cell and provide specific immunity. Chimeric antigen receptor-expressing T cells may be used in various therapies, including cancer therapies. Costimulating polypeptides may be used to enhance the activation of CAR-expressing T cells against target antigens, and therefore increase the potency of adoptive immunotherapy. Brentjens et al. describe the use of a second generation of a CD19 CAR, expressing the CD28 co-stimulatory domain and CD3ζ for use in the treatment of adults with relapsed B-acute lymphoblastic leukaemia (Brentjens et al., Sci. Transl. Med. 5, 177ra38-177ra38, 2013). WO 2010/033949 A1 discloses a method for activating an antigen-presenting cell, comprising a cytoplasmic CD40 polypeptide region and a MyD88 peptide or truncated version thereof. Further, WO 2015/123527 A1 describes the co-expression of a co-stimulatory MyD88/CD40 molecule together with a CAR has been disclosed. Additionally, WO 2014/151960 A2 relates to the transduction of T-cells with an inducible MyD88/CD40 construct in combination or alone with a CAR. This construct comprises an Fv'Fv' sequence for dimerization and activation. Summary The present invention is defined by the claims. Accordingly the present invention relates in a first aspect to a nucleic acid comprising a promoter operably linked to a polynucleotide encoding a cytoplasmic chimeric stimulating protein, wherein the cytoplasmic chimeric stimulating protein 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; with the proviso that the cytoplasmic chimeric stimulating protein does not include a multimerizing or dimerizing ligand binding site. In accordance with a preferred embodiment the cytoplasmic chimeric stimulating protein is constitutively active. In accordance with a preferred embodiment the nucleic acid further comprises a second polynucleotide encoding a chimeric antigen receptor, or a T cell receptor. In accordance with a preferred embodiment the second polynucleotide encodes a chimeric antigen receptor comprising a transmembrane region, a T cell activation molecule, and an antigen recognition moiety. In accordance with a preferred embodiment the chimeric antigen receptor is encoded 5' of the cytoplasmic chimeric stimulating protein. In accordance with a preferred embodiment one promoter is operably linked to both the first and second polynucleotides. In accordance with a preferred embodiment the nucleic acid comprises a polynucleotide encoding a 2A polypeptide linker between the first polynucleotide and the second polynucleotide. The present invention relates in a second aspect to a nucleic acid comprising a promoter operably linked to a polynucleotide encoding a chimeric antigen receptor polypeptide, wherein the chimeric antigen receptor polypeptide 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. In accordance with a preferred embodiment the nucleic acid comprises a polynucleotide that encodes a chimeric Caspase-9 polypeptide comprising a multimeric ligand binding region and a Caspase-9 polypeptide. The