Compositions and methods for generating an immune response to a tumor associated antigen
The compositions and methods are described for generating an immune response to a tumor associated antigen such as MUC1. The compositions and methods described herein relate to a modified vaccinia Ankara (MVA) vector encoding one or more viral antigens for generating a protective immune response to a neoplasm expressing the tumor associated antigen in the subject to which the vector is administered. The compositions and methods of the present invention are useful both prophylactically and therapeutically and may be used to prevent and/or treat neoplasms and associated diseases.
1. A recombinant modified vaccinia ankara (MVA) viral vector comprising:
(i) a first nucleic acid sequence encoding a chimeric amino acid sequence comprising (a) an extracellular fragment of a tumor associated antigen (TAA), (b) a transmembrane domain of a glycoprotein (GP) of Marburg virus, and (c) an intracellular fragment of the TAA; and
(ii) a second nucleic acid sequence encoding a Marburg virus VP40 matrix protein; wherein both the first nucleic acid sequence and the second nucleic acid sequence are under the control of promoters compatible with poxvirus expression systems, and wherein upon expression, the chimeric amino acid sequence and VP40 matrix protein are capable of assembling together to form virus like particles (VLPs).
2. The recombinant MVA viral vector of claim 1 , wherein the first nucleic acid sequence and the second nucleic acid sequence are inserted into one or more deletion sites of the MVA selected from I, II, III, IV, V or VI.
3. The recombinant MVA viral vector of claim 1 , wherein the first nucleic acid sequence and the second nucleic acid sequence are inserted into the MVA in a natural deletion site, a modified natural deletion site, or between essential or non-essential MVA genes.
4. The recombinant MVA viral vector of claim 1 , wherein the first nucleic acid sequence and the second nucleic acid sequence are inserted into the same natural deletion site, a modified natural deletion site, or between the same essential or non-essential MVA genes.
5. The recombinant MVA viral vector of claim 1 , wherein the first nucleic acid sequence and the second nucleic acid sequence are inserted into different natural deletion sites, different modified deletion sites, or between different essential or non-essential MVA genes.
6. The recombinant MVA viral vector of claim 1 , wherein the first nucleic acid sequence is inserted between two essential and highly conserved MVA genes; and the second nucleic acid sequence is inserted into a restructured and modified deletion III insertion site.
7. The recombinant MVA viral vector of claim 1 , wherein the first nucleic acid sequence is inserted between MVA genes I8R and GIL.
8. The recombinant MVA viral vector of claim 1 , wherein the promoter is Pm2H5, Psyn II, or mH5 promoters or combinations thereof.
9. The recombinant MVA viral vector of claim 1 , wherein the TAA sequence is optimized by one or more methods selected from the group consisting of changing selected codons to other synonymous codons that are optimal for protein expression by MVA, interrupting homopolymer stretches using silent mutations, and interrupting transcription terminator motifs using silent mutations.
10. The recombinant MVA viral vector of claim 1 , wherein the chimeric amino acid sequence comprises (a) an extracellular fragment of mucin-1 (MUC-1), (b) a transmembrane domain of a glycoprotein (GP) of Marburg virus, and (c) an intracellular fragment of MUC-1.
11. The recombinant MVA viral vector of claim 10 , wherein the extracellular fragment comprises the amino acid sequence of SEQ ID NO:1-or SEQ ID NO:2.
12. The recombinant MVA viral vector of claim 10 , wherein the extracellular fragment comprises the amino acid sequence of SEQ ID NO:4.
13. The recombinant MVA viral vector of claim 10 , wherein the extracellular fragment comprises the amino acid sequence of SEQ ID NO:2.
14. The recombinant MVA viral vector of claim 10 , wherein the extracellular fragment comprises the amino acid sequence of SEQ ID NO:3.
15. The recombinant MVA viral vector of claim 10 , wherein the intracellular fragment of MUC-1 comprises amino acids 407-475 of SEQ ID NO: 6.
16. The recombinant MVA viral vector of claim 1 , wherein the transmembrane domain of the GP of Marburg virus comprises the amino acid sequence of SEQ ID NO:13.
17. A pharmaceutical composition comprising at least one recombinant MVA viral vector of claim 1 and a pharmaceutically acceptable carrier.
18. The pharmaceutical composition of claim 17 , wherein the at least one recombinant MVA viral vector is formulated for intraperitoneal, intramuscular, intradermal, epidermal, mucosal or intravenous administration.
19. The pharmaceutical composition of claim 17 , wherein the recombinant MVA viral vector comprises a chimeric amino acid sequence comprising (a) an extracellular fragment of MUC-1, (b) a transmembrane domain of a glycoprotein (GP) of Marburg virus, and (c) an intracellular fragment of MUC-1.