Cell-based neoantigen vaccines and uses thereof
The present disclosure relates to immune cells that express an exogenous neoantigen and an immunogenicity enhancer, or to T cells that express an exogenous neoantigen, and their use in treating a disease or disorder, such as cancer for tumor associated neoantigens. Related expression constructs, kits, host cells, pharmaceutical compositions, and methods are also provided.
1. A T cell comprising:
a heterologous polynucleotide encoding a fusion protein, wherein the encoded fusion protein comprises:
(i)(a) a tumor neoantigen, wherein the tumor neoantigen comprises a peptide that binds a human Class I Major Histocompatibility Complex (MHC) molecule; and
(i)(b) an antigen peptide or tumor neoantigen peptide, wherein the antigen peptide or tumor neoantigen peptide binds a human Class II MHC molecule; and
(ii) a polynucleotide encoding an immunogenicity enhancer.
2. The T cell of claim 1 , wherein the encoded fusion protein comprises (i)(a) the tumor neoantigen comprising a peptide that binds a human Class I MHC molecule and (i)(b) a tumor neoantigen that binds a human Class II MHC molecule.
3. The T cell of claim 1 , wherein the T cell encodes a plurality of tumor neoantigens.
4. The T cell of claim 1 , wherein the immunogenicity enhancer comprises an interleukin-12 (IL-12), a granulocyte-macrophage colony-stimulating factor (GM-CSF), an inducible cell death factor, a bacterial flagellin, a CD80, a CD137L, a CD40L, a secreted interleukin-2 (IL-2), a secreted IL-2 that binds T cells independent of CD25, a secreted interleukin-15 (IL-15), a secreted IL-15-IL-15Rα complex, a secreted interferon-β (IFNβ), a secreted interferon-α1 (IFN-α1), a secreted interleukin-7 (IL-7), or any combination thereof.
5. The T cell of claim 4 , wherein the immunogenicity enhancer comprises:
(a) a fusion protein that localizes to the cell surface of the T cell and comprises the IL-12;
(b) a fusion protein that localizes to the cell surface of the T cell and comprises the IL-12, and the GM-CSF;
(c) the IL-12 and the GM-CSF;
(d) the inducible cell death factor, wherein the inducible cell death factor is comprised of a receptor interacting serine/threonine kinase 3 (RIPK3);
(e) a fusion protein comprising the inducible cell death factor and a multimerization domain, wherein the inducible cell death factor comprises a RIPK3 kinase domain and the multimerization domain comprises a FK506-binding protein or multimerizing portion thereof;
(f) the bacterial flagellin, wherein the bacterial flagellin is comprised of a Salmonella phase 1 flagellin;
(g) a fusion protein that localizes to the cell surface of the T cell and comprises the bacterial flagellin; or
(h) the GM-CSF.
6. The T cell of claim 4 , wherein the immunogenicity enhancer comprises CD80, CD137L, IFN-β, IL-12, GM-CSF, or any combination thereof.
7. The T cell of claim 1 , further comprising a heterologous polynucleotide encoding a costimulatory molecule.
8. The T cell of claim 7 , wherein the costimulatory molecule comprises a CD80, a CD86, a B7RP1, a CD137L, an OX40L, a CD70, a CD30L, a CD154, an ICAM-1, a CD2BP2, a LIGHT, a KLRD1, a ligand that specifically binds to a CD83, an agonist of CD137 (4-1BB), an agonist of CD134 (OX-40), an agonist of CD27, an agonist of CD28, an agonist of CD40, an agonist of CD122, an agonist of GITR, an agonist of ICOS, or any combination thereof.
9. A composition comprising the T cell of claim 1 and a pharmaceutically acceptable carrier, diluent, or excipient.
10. The T cell of claim 1 , wherein:
(i) the T cell is a naïve, central memory, naïve and central memory, effector memory, or combination thereof;
(ii) the T cell is a CD4+ T cell, a CD8+ T cell, or both;
(iii) the T cell is a human T cell; or
(iv) any combination of (i)-(iii).
11. The T cell of claim 1 , wherein the polynucleotide encoding the fusion protein is comprised in a transposon expression construct that comprises one or more minigenes each encoding a tumor neoantigen.
12. The T cell of claim 1 , wherein the fusion protein comprises an antigen from KRAS, B-Raf, SF31, MYD88, DDX3X, MAPK1, GNB1, p53, raf, ras, myc, pRb, PTEN, CD95, α-fetoprotein (AFP), B7H4, BTLA, CD3, CD19, CD20, CD25, CD22, CD28, CD30, CD40, CD44v6, CD52, CD56, CD79b, CD80, CD81, CD86, CD134 (OX40), CD137 (4-1BB), CD151, CD276, CA125, CEA, CEACAM6, 5 c-Met, CT-7, CTLA-4, EGFR, EGFRvIII, ErbB2, ErbB3, ErbB4, EphA2, FLT1, FLT4, Frizzled, O-acetyl-GD2, GD2, GHRHR, GHR, GITR, gp130, HVEM, IGF1R, IL6R, KDR, L1CAM, Lewis A, Lewis Y, LTβR, LIFRβ, LRP5, MAGE, mesothelin, MUC1, NY-ESO-1, OSMRβ, PD1, PD-L1, PD-L2, PSMA, PTCH1, RANK, Robo1, ROR1, TERT, TGFBR2, TGFBR1, TLR7, TLR9, TNFRSF4, 10 TNFR1, TNFR2, tyrosinase, TWEAK-R, or WT-1.
13. The T cell of claim 1 , wherein the immunogenicity enhancer, when expressed by the T cell, is capable of improving an immune response against the tumor neoantigen or tumor neoantigens as compared to the T cell encoding the tumor neoantigen or tumor neoantigens without the immunogenicity enhancer, wherein the immune response comprises direct and/or indirect activation of T cells, wherein indirect activation is mediated by dendritic cells.
14. The T cell of claim 1 , wherein the fusion protein comprises a peptide linker disposed between and connecting:
(1) two tumor neoantigens; and/or
(2) a tumor neoantigen comprising the peptide that binds a human Class I MHC molecule and an antigen peptide or tumor neoantigen peptide that binds a human Class II MHC molecule.
15. A human T cell comprising:
a heterologous polynucleotide encoding a fusion protein, wherein the encoded fusion protein comprises:
(i)(a) cancer-associated neoantigen, wherein the cancer-associated neoantigen comprises a peptide that binds a human Class I Major Histocompatibility Complex (MHC) molecule; and
(i)(b) an antigen peptide or cancer-associated neoantigen peptide, wherein the antigen peptide or neoantigen peptide binds a human Class II MHC molecule; and
(ii) a polynucleotide encoding an immunogenicity enhancer.
16. The human T cell of claim 15 , wherein the T cell encodes a plurality of cancer-associated neoantigens.
17. The human T cell of claim 15 , wherein the immunogenicity enhancer comprises CD80, CD137L, IFN-β, IL-12, GM-CSF, or any combination thereof.
18. The human T cell of claim 15 , wherein the fusion protein comprises a peptide linker disposed between and connecting:
(1) two cancer-associated neoantigens; and/or
(2) a cancer-associated neoantigen comprising a peptide that binds a human Class I MHC molecule and an antigen peptide or cancer-associated neoantigen peptide that binds a human Class II MHC molecule.
19. A human T cell comprising:
a heterologous polynucleotide encoding a fusion protein, wherein the encoded fusion protein comprises:
(i)(a) cancer-associated neoantigen, wherein the cancer-associated neoantigen comprises a peptide that binds a human Class II Major Histocompatibility Complex (MHC) molecule; and
(i)(b) an antigen peptide or cancer-associated neoantigen peptide, wherein the antigen peptide or cancer-associated neoantigen peptide binds a human Class I MHC molecule; and
(ii) a polynucleotide encoding an immunogenicity enhancer.
20. The human T cell of claim 19 , wherein the immunogenicity enhancer comprises CD80, CD137L, IFN-β, IL-12, GM-CSF, or any combination thereof.
21. The human T cell of claim 19 , wherein the encoded fusion protein comprises a peptide linker disposed between and connecting:
(1) two cancer-associated neoantigens; and/or
(2) (2)(a) a cancer-associated neoantigen comprising a peptide that binds a human Class II MHC molecule and (2)(b) an antigen peptide or cancer-associated neoantigen peptide that binds a human Class I MHC molecule.
22. The human T cell of claim 19 , wherein the T cell encodes a plurality of cancer-associated neoantigens.