Compositions and methods for enhancing the stability of transgenes in poxviruses
Provided herein are recombinant poxviruses that are stable through successive passaging of the recombinant poxviruses. More particularly, the recombinant poxviruses comprise one or more modified nucleic acids encoding MUC1, CEA, and/or TRICOM antigens, wherein the recombinant poxviruses are stable through successive passaging. Also, provided herein are compositions and method related thereto.
1. A recombinant poxvirus which is stable through successive passaging of the recombinant poxvirus, the recombinant poxvirus comprising a nucleic acid encoding a MUC1 peptide having three Variable N-Terminal Repeat (VNTR) domains, wherein (a) the order of the nucleic acids encoding at least two VNTR domains of MUC1 is different from their order in PANVAC, and (b) the VNTR domains are codon optimized, wherein the recombinant poxvirus is stable through successive passaging of the recombinant poxvirus.
2. The recombinant poxvirus of claim 1 , wherein said nucleic acid sequence comprises SEQ ID NO:2.
3. The recombinant poxvirus of claim 1 , further comprising a nucleic acid encoding a carcinoembryonic antigen (CEA).
4. The recombinant poxvirus of claim 3 , wherein the nucleic acid encoding a carcinoembryonic antigen (CEA) comprises at least one nucleotide substitution in at least one repetitive nucleotide region, wherein the at least one repetitive nucleotide region is defined as (a) three or more consecutively repeated G or C nucleotides and/or (b) three or more consecutively repeated T nucleotides.
5. The recombinant poxvirus of claim 4 , wherein the nucleic acid encoding a carcinoembryonic antigen (CEA) comprises SEQ ID NO:13 or SEQ ID NO: 14.
6. The recombinant poxvirus of claim 1 , wherein the recombinant poxvirus is selected from an orthopoxvirus and an avipoxvirus.
7. The recombinant poxvirus of claim 6 , wherein the orthopoxvirus is a modified vaccinia virus Ankara (MVA) that is MVA-BN.
8. The recombinant poxvirus of claim 1 , wherein the nucleic acid encoding a MUC1 peptide further comprises at least one nucleotide sequence encoding a peptide fragment selected from the group consisting of:
(SEQ ID NO: 24)
YLAPPAHGV,
(SEQ ID NO: 25)
YLDTRPAPV,
(SEQ ID NO: 26)
YLAIVYLIAL,
(SEQ ID NO: 27)
YLIALAVCQV,
(SEQ ID NO: 28)
YLSYTNPAV,
and
(SEQ ID NO: 29)
SLFRSPYEK.
9. The recombinant poxvirus of claim 1 , wherein the poxvirus further comprises a nucleic acid encoding one or more co-stimulatory molecules selected from B7-1, ICAM-1, LFA-3, and combinations thereof.
10. A host cell comprising the recombinant poxvirus of claim 1 .
11. A composition comprising the recombinant poxvirus of claim 1 that is suitable for administration as part of a homologous prime-boost regimen or a heterologous prime-boost regimen.
12. The recombinant poxvirus of claim 1 that comprises a nucleic acid encoding a MUC1 peptide in which the order of VNTR #2 and VNTR #1 is reversed.
13. The recombinant poxvirus of claim 1 that has stability through passage 7.