Tri-segmented arenaviruses as vaccine vectors
The present application relates to arenaviruses with rearrangements of their open reading frames (“ORF”) in their genomes. In particular, described herein is a modified arenavirus genomic segment, wherein the arenavirus genomic segment is engineered to carry a viral ORF in a position other than the wild-type position of the ORF. Also described herein are trisegmented arenavirus particles comprising one L segment and two S segments or two L segments and one S segment. The arenavirus, described herein may be suitable for vaccines and/or treatment of diseases and/or for the use in immunotherapies.
1. An arenavirus genomic segment, wherein the genomic segment is engineered to carry a viral open reading frame (“ORF”) in a position other than the wild-type position of the ORF, wherein the arenavirus genomic segment is selected from the group consisting of:
(i) an S segment, wherein the ORF encoding the nucleoprotein (“NP”) is under control of an arenavirus 5′ untranslated region (“UTR”);
(ii) an S segment, wherein the ORF encoding the matrix protein Z (“Z protein”) is under control of an arenavirus 5′ UTR;
(iii) an S segment, wherein the ORF encoding the RNA dependent RNA polymerase L (“L protein”) is under control of an arenavirus 5′ UTR;
(iv) an S segment, wherein the ORF encoding the viral glycoprotein (“GP”) is under control of an arenavirus 3′ UTR;
(v) an S segment, wherein the ORF encoding the L protein is under control of an arenavirus 3′ UTR; and
(vi) an S segment, wherein the ORF encoding the Z protein is under control of an arenavirus 3′ UTR.
2. The arenavirus genomic segment of claim 1 , wherein the arenavirus 3′ UTR is the 3′ UTR of the arenavirus S segment, and wherein the arenavirus 5′ UTR is the 5′ UTR of the arenavirus S segment.
3. A cDNA of the arenavirus genomic segment of claim 1 .
4. A DNA expression vector comprising the cDNA of claim 3 .
5. A host cell comprising the vector of claim 4 .
6. An arenavirus particle comprising the arenavirus genomic segment of claim 1 and a second arenavirus genomic segment so that the arenavirus particle comprises an S segment and an L segment.
7. The arenavirus particle of claim 6 , wherein the arenavirus particle is:
(i) infectious and replication competent;
(ii) attenuated; or
(iii) infectious but unable to produce further infectious progeny in non-complementing cells.
8. The arenavirus particle of claim 7 , wherein:
(i) at least one of the four ORFs encoding GP, NP, Z protein, and L protein is removed or functionally inactivated;
(ii) at least one of the four ORFs encoding GP, NP, Z protein, and L protein is removed and replaced with a heterologous ORF from an organism other than an arenavirus;
(iii) only one of the four ORFs encoding GP, NP, Z protein and L protein is removed and replaced with a heterologous ORF from an organism other than an arenavirus;
(iv) the ORF encoding GP is removed and replaced with a heterologous ORF from an organism other than an arenavirus;
(v) the ORF encoding NP is removed and replaced with a heterologous ORF from an organism other than an arenavirus;
(vi) the ORF encoding the Z protein is removed and replaced with a heterologous ORF from an organism other than an arenavirus; or
(vii) the ORF encoding the L protein is removed and replaced with a heterologous ORF from an organism other than an arenavirus.
9. The arenavirus particle of claim 8 , wherein the heterologous ORF encodes an antigen derived from an infectious organism, tumor, or allergen.
10. The arenavirus particle of claim 9 , wherein the heterologous ORF encoding an antigen is selected from human immunodeficiency virus antigens, hepatitis C virus antigens, varizella zoster virus antigens, cytomegalovirus antigens, Mycobacterium tuberculosis antigens, and tumor associated antigens.
11. A vaccine comprising the arenavirus particle of claim 8 and a pharmaceutically acceptable carrier.
12. A pharmaceutical composition comprising an arenavirus particle of claim 8 and a pharmaceutically acceptable carrier.
13. The arenavirus genomic segment of claim 1 , wherein the arenavirus genomic segment is derived from lymphocytic choriomeningitis virus (“LCMV”).
14. A tri-segmented arenavirus particle comprising one L segment and two S segments, wherein one of the two S segments is selected from the group consisting of:
(i) an S segment, wherein the ORF encoding the NP is under control of an arenavirus 5′ UTR,
(ii) an S segment, wherein the ORF encoding the Z protein is under control of an arenavirus 5′ UTR,
(iii) an S segment, wherein the ORF encoding the L protein is under control of an arenavirus 5′ UTR,
(iv) an S segment, wherein the ORF encoding the GP is under control of an arenavirus 3′ UTR,
(v) an S segment, wherein the ORF encoding the L protein is under control of an arenavirus 3′ UTR, and
(vi) an S segment, wherein the ORF encoding the Z protein is under control of an arenavirus 3′ UTR.
15. The tri-segmented arenavirus particle of claim 14 , wherein inter-segmental recombination of the two S segments, uniting two arenavirus ORFs on only one instead of two separate segments, abrogates viral promoter activity.
16. The tri-segmented arenavirus particle of claim 14 , wherein one of the two S segments is
an S segment, wherein the ORF encoding the GP is under control of an arenavirus 3′ UTR.
17. The tri-segmented arenavirus particle of claim 16 , wherein the arenavirus 3′ UTR is the 3′ UTR of the arenavirus S segment, and wherein the arenavirus 5′ UTR is the 5′ UTR of the arenavirus S segment.
18. The tri-segmented arenavirus particle of claim 14 , wherein the two S segments comprise (i) one or two heterologous ORFs from an organism other than an arenavirus; or (ii) one or two duplicated arenavirus ORFs; or (iii) one heterologous ORF from an organism other than an arenavirus and one duplicated arenavirus ORF.
19. The tri-segmented arenavirus particle of claim 18 , wherein the heterologous ORF encodes an antigen derived from an infectious organism, tumor, or allergen.
20. The tri-segmented arenavirus particle of claim 19 , wherein the heterologous ORF encoding an antigen is selected from human immunodeficiency virus antigens, hepatitis C virus antigens, varizella zoster virus antigens, cytomegalovirus antigens, Mycobacterium tuberculosis antigens, and tumor associated antigens.
21. The tri-segmented arenavirus particle of claim 14 , wherein the tri-segmented arenavirus particle comprises all four arenavirus ORFs, and wherein the tri-segmented arenavirus particle is infectious and replication competent.
22. A tri-segmented arenavirus particle comprising one L segment and two S segments, wherein a first S segment is engineered to carry an ORF encoding GP in a position under control of an arenavirus 3′ UTR and an ORF encoding a first gene of interest in a position under control of an arenavirus 5′ UTR and a second S segment is engineered to carry an ORF encoding NP in a position under control of an arenavirus 3′ UTR and an ORF encoding a second gene of interest in a position under control of an arenavirus 5′ UTR.
23. A tri-segmented arenavirus particle comprising one L segment and two S segments, wherein a first S segment is engineered to carry an ORF encoding GP in a position under control of an arenavirus 5′ UTR and an ORF encoding a first gene of interest in a position under control of an arenavirus 3′ UTR and a second S segment is engineered to carry an ORF encoding NP in a position is under control of an arenavirus 5′ UTR and an ORF encoding a second gene of interest in a position under control of an arenavirus 3′ UTR.
24. The tri-segmented arenavirus particle of claim 22 or 23 , wherein the gene of interest encodes an antigen derived from an infectious organism, tumor, or allergen.
25. The tri-segmented arenavirus particle of claim 24 , wherein the gene of interest encodes an antigen selected from human immunodeficiency virus antigens, hepatitis C virus antigens, varizella zoster virus antigens, cytomegalovirus antigens, Mycobacterium tuberculosis antigens, and tumor associated antigens.
26. A cDNA of the tri-segmented arenavirus particle genome of claim 14 or 22 or 23 .
27. A DNA expression vector comprising the cDNA of claim 26 .
28. A host cell comprising the vector of claim 27 .
29. A vaccine comprising a tri-segmented arenavirus particle of any one of claim 14 , 22 , or 23 and a pharmaceutically acceptable carrier.
30. A pharmaceutical composition comprising a tri-segmented arenavirus particle of any one of the claims 14 , 22 , or 23 and a pharmaceutically acceptable carrier.
31. The tri-segmented arenavirus particle of any one of claim 14 , 22 , or 23 , wherein the tri-segmented arenavirus particle is derived from LCMV.