Methods and compositions for transgene expression in a herpesvirus vector system
Compositions are provided comprising a replication-competent herpesvirus vector system and/or a herpesvirus vector particle comprising a heterologous recombinant transgene of interest encoding an antigen of interest (e.g., a tumor-associated antigen, a microbial antigen, etc.), wherein the recombinant transgene has been modified to comprise a codon usage signature of a herpesvirus late gene operably linked to an active promoter. Such compositions find use in a variety of methods including, for example, methods of generating an immune response against an antigen of interest in a subject in need thereof, as well as methods for treating or preventing cancer or a microbial infection.
1. A replication-competent herpesvirus vector system comprising (a) a polynucleotide encoding a trans-inducer polypeptide that enhances expression of a herpesvirus late gene, and (b) a polynucleotide comprising a heterologous recombinant transgene of interest operably linked to a promoter, wherein said heterologous recombinant transgene of interest comprises a codon usage signature of the herpesvirus late gene.
2. The replication-competent herpesvirus vector system of claim 1 , wherein said herpesvirus vector system is derived from a Gammaherpes virus.
3. The replication-competent herpesvirus vector system of claim 2 , wherein said Gammaherpesvirus is a gamma-2 herpesvirus.
4. The replication-competent herpesvirus vector system of claim 1 , wherein said herpesvirus vector system is derived from a herpesvirus of the Betaherpesvirinae subfamily.
5. The replication-competent herpesvirus vector system of claim 1 , wherein said herpesvirus vector system is derived from a herpesvirus of the Alphaherpesvirinae subfamily.
6. The replication-competent herpesvirus vector system of claim 1 , wherein said heterologous recombinant transgene of interest encodes an antigen.
7. The replication-competent herpesvirus vector system of claim 6 , wherein said antigen is a microbial antigen or a tumor-associated antigen.
8. The replication-competent herpesvirus vector system of claim 7 , wherein said microbial antigen comprises a viral antigen, a parasitic antigen, a mycobacteria antigen, or a bacterial antigen.
9. The replication-competent herpesvirus vector system of claim 8 , wherein said viral antigen comprises a viral antigen from Retroviridae or Flaviviridae.
10. The replication-competent herpesvirus vector system of claim 9 , wherein said Retroviridae or Flaviviridae viral antigen comprises a viral antigen from a dengue virus, a lentivirus, a human immunodeficiency virus (HIV) or a simian immunodeficiency virus (SIV).
11. The replication-competent herpesvirus vector system of claim 8 , wherein said viral antigen comprises an envelope polypeptide.
12. The replication-competent herpesvirus vector system of claim 11 , wherein said envelope polypeptide is from HIV or SIV.
13. The replication-competent herpesvirus vector system of claim 12 , wherein said recombinant transgene comprises the polynucleotide set forth in SEQ ID NO: 1 or a polynucleotide having at least 90% sequence identity to SEQ ID NO: I, wherein said polynucleotide provides an improved immune response when compared to a recombinant transgene that encodes the envelope polypeptide from HIV or SIV but that lacks the codon usage signature.
14. The replication-competent herpesvirus vector system of claim 8 , wherein said bacterial antigen is from Mycobacterium tuberculosis.
15. The replication-competent herpesvirus vector system of claim 7 , wherein said microbial antigen is from Plasmodium.
16. The replication-competent herpesvirus vector system of claim 7 , wherein said tumor-associated antigen (a) is associated with melanoma, lymphoma, leukemia, lung cancer, bladder cancer, colon cancer, breast cancer, prostate cancer, esophageal cancer, liver cancer, pancreatic cancer, ovarian cancer, non-small cell lung cancer, renal cancer, neuroblastoma, colorectal cancer, uterine cancer, acute myelocytic leukemia, sarcoma, brain cancer or bone cancer or (b) comprises BAGE, GAGE, MAGE, NY-ESO-1, SSX, gp100, Melan-A/Mart-1, Tyrosinase, PSA, CEA, Mammaglobin-A, p53, HER-2/neu, livin, survivin, B-catenin-m, B-Actin/4/m, Myosin/m, HSP70-2/m, HLA-A2-R170J, GM2, GD2, GD3, MUC-1, sTn, globo-H, WT1, PR1, E75, ras, AFP, URLC10, VEGFR1 and 2, mutant p53, NY-ESO-1, HPV16 E7, B-catenin, CDK4, CDC27, a actinin-4, TRPI/gp75, TRP2, gangliosides, PSMA, HER2, WT1, EphA3, EGFR, CD20, telomerase, MART-1, or an antigenic portion thereof.
17. A host cell or cell line comprising the replication-competent herpesvirus vector system of claim 1 .
18. A replication-competent herpesvirus vector particle comprising within its genome a polynucleotide comprising a sequence encoding a trans-inducer polypeptide that enhances expression of a herpesvirus late gene and a heterologous recombinant transgene of interest operably linked to a promoter, wherein said heterologous recombinant transgene comprises a codon usage signature of the herpesvirus late gene.
19. The replication-competent herpesvirus vector particle of claim 18 , wherein said herpesvirus particle is derived from a Gammaherpes virus.
20. The replication-competent herpesvirus vector particle of claim 19 , wherein said Gammaherpes virus is a gamma-2 herpesvirus.
21. The replication-competent herpesvirus vector particle of claim 18 , wherein said herpesvirus particle is derived from a herpesvirus of the Betaherpesvirinae subfamily.
22. The replication-competent herpesvirus vector particle of claim 18 , wherein said herpesvirus particle is derived from a herpesvirus of the Alphaherpesvirinae subfamily.
23. The replication-competent herpesvirus vector particle of claim 18 , wherein said heterologous recombinant transgene of interest encodes an antigen.
24. The replication-competent herpesvirus vector particle of claim 23 , wherein said antigen is a microbial antigen or a tumor-associated antigen.
25. The replication-competent herpesvirus vector particle of claim 24 , wherein said microbial antigen comprises a viral antigen, a parasitic antigen, a mycobacteria antigen or a bacterial antigen.
26. A pharmaceutical composition comprising the replication-competent herpesvirus vector particle of claim 18 .
27. A method for delivering a transgene of interest to a cell comprising contacting the cell with a replication-competent herpesvirus vector particle of claim 18 .
28. A method of generating an immune response against a microbial antigen in a subject in need thereof comprising administering to said subject a therapeutically effective amount of the replication competent herpesvirus vector particle of claim 18 , wherein the heterologous recombinant transgene encodes the microbial antigen, and wherein an immune response against said microbial antigen is generated in said subject.
29. A method for treating or inhibiting a microbial infection or cancer in a subject in need thereof comprising administering to said subject a therapeutically effective amount of the replication-competent herpesvirus vector particle of claim 18 , wherein the heterologous recombinant transgene encodes a microbial antigen or a tumor-associated antigen, and wherein an immune response against said microbial antigen or tumor-associated antibody is generated in said subject and thereby treats or inhibits said microbial infection or cancer.
30. The replication-competent herpesvirus vector system of claim 1 , wherein the herpesvirus late gene comprises a polynucleotide encoding a capsid protein, a tegument protein, or an envelope protein.
31. The replication-competent herpesvirus vector system of claim 1 , wherein the herpesvirus late gene comprises a polynucleotide encoding a glycoprotein.
32. The replication-competent herpesvirus vector system of claim 31 , wherein the glycoprotein is selected from the group consisting of glycoprotein B (gB), glycoprotein C (gC), glycoprotein D (gD), glycoprotein H (gH), and glycoprotein L (gL).
33. The replication-competent herpesvirus vector system of claim 1 , wherein the trans-inducer polypeptide is ORF57.
34. A method of increasing expression of a transgene by a host cell, comprising (a) altering codons of the transgene to reflect the codon usage signature of a herpesvirus late gene, (b) inserting the transgene with the altered codons into a replication-competent herpesvirus vector, (c) introducing into the host cell the replication-competent herpesvirus vector, and (d) culturing the host cell comprising the replication-competent herpesvirus vector in the presence of a herpesvirus trans-inducer polypeptide that enhances expression of the herpesvirus late gene.
35. The method of claim 34 , wherein the replication-competent herpesvirus vector is derived from a Gammaherpes virus, a Betaherpes virus, or an Alphaherpes virus.
36. The method of claim 34 , wherein the herpesvirus late gene comprises a polynucleotide encoding a capsid protein, a tegument protein, or an envelope protein.
37. The method of claim 34 , wherein the herpesvirus late gene comprises a polynucleotide encoding a glycoprotein.
38. The method of claim 37 , wherein the glycoprotein is selected from the group consisting of glycoprotein B (gB), glycoprotein C (gC), glycoprotein D (gD), glycoprotein H (gH), and glycoprotein L (gL).
39. The method of claim 38 , wherein the trans-inducer polypeptide is ORF57.
40. The method of claim 34 , comprising altering codons of the transgene to codons that are present at high frequency in the coding sequence of the herpesvirus late gene.
41. The method of claim 34 , wherein the altered codons are codons that are present at high frequency in the coding sequence of the herpesvirus late gene.
42. The replication-competent herpesvirus vector system of claim 1 , wherein said heterologous recombinant transgene of interest comprises codons used at high frequency in the coding sequence of the herpesvirus late gene.
43. The replication-competent herpesvirus vector system of claim 1 , wherein said heterologous recombinant transgene of interest has been modified to comprise one or more codons present at high frequency in the coding sequence of the herpesvirus late gene.