Alphavirus neoantigen vectors
Disclosed herein are alphavirus vectors that include neoantigen-encoding nucleic acid sequences derived from a tumor of a subject. Also disclosed are nucleotides, cells, and methods associated with the vectors including their use as vaccines.
1. A method for inducing an immune response in a subject, comprising:
(A) administering a first vaccine composition, wherein the first vaccine composition comprises a chimpanzee adenovirus vector comprising a first neoantigen cassette; and
(B) administering a second vaccine composition comprising an neoantigen expression system, wherein the neoantigen expression system comprises one or more vectors, wherein the one or more vectors each comprise:
(a) an RNA alphavirus backbone, wherein the RNA alphavirus backbone comprises:
(i) at least one promoter nucleotide sequence, and
(ii) at least one polyadenylation (poly(A)) sequence; and
(b) a second neoantigen cassette; and
wherein the first neoantigen cassette and the second neoantigen cassette comprise
at least one neoantigene-encoding nucleic acid sequence that is the same.
2. The method of claim 1 , wherein the first neoantigen cassette and the second neoantigen cassette comprise the same neoantigene-encoding nucleic acid sequences.
3. The method of claim 1 , wherein the second neoantigen cassette comprises at least one neoantigene-encoding nucleic acid sequence that is different with respect to any neoantigene-encoding nucleic acid sequences present in the first neoantigen cassette.
4. The method of claim 1 , wherein the first vaccine composition is administered prior to the second vaccine composition.
5. The method of claim 1 , wherein the first vaccine composition is administered is administered as a priming dose.
6. The method of claim 1 , wherein the RNA alphavirus backbone comprises:
(a) the sequence of SEQ ID NO:3 further comprising a deletion between base pair 7544 and 11175, wherein the second neoantigen cassette is inserted to replace the deletion between base pair 7544 and 11175, and/or
(b) the sequence set forth in SEQ ID NO:6, wherein the second neoantigen cassette is inserted at position 7544 of the sequence set forth in SEQ ID NO:6.
7. The method of claim 1 , wherein one or both of the first neoantigen cassette and the second neoantigen cassette comprise junctional epitope sequences encoded by adjacent neoantigen-encoding nucleic acid sequences in the respective neoantigen cassettes, wherein at least one or each junctional epitope sequence has an affinity of greater than 500 nM for MHC, and optionally wherein at least one or each junctional epitope sequence is non-self.
8. The method of claim 1 , wherein the chimpanzee adenovirus vector comprises a ChAdV68-based vector.
9. The method of claim 1 , wherein one or both of the first neoantigen cassette and the second neoantigen cassette comprise at least two neoantigen-encoding nucleic acid sequences, wherein each neoantigen-encoding nucleic acid sequence comprises an epitope-encoding nucleic acid sequence, and
wherein each of the at least two neoantigen-encoding nucleic acid sequences within the first neoantigen cassette and the second neoantigen cassette, respectively, is linked directly to one another.
10. The method of claim 9 , wherein each neoantigen-encoding nucleic acid sequence comprises:
(A) the epitope-encoding nucleic acid sequence corresponding to the respective neoantigen-encoding nucleic acid sequence;
(B) a 5′ linker sequence that encodes a N-terminal amino acid sequence, and wherein the 5′ linker sequence encodes a peptide that is between 2-20 amino acids in length; and
(C) a 3′ linker sequence that encodes a C-terminal amino acid sequence, and wherein the 3′ linker sequence encodes a peptide that is between 2-20 amino acids in length.
11. The method of claim 10 , wherein:
(A) the 5′ linker sequence comprises a native 5′ linker sequence that encodes a native N-terminal amino acid sequence of the epitope encoded by the epitope-encoding nucleic acid sequence, and
(B) the 3′ linker sequence comprises a native 3′ linker sequence that encodes a native C-terminal amino acid sequence of the epitope encoded by the epitope-encoding nucleic acid sequence.
12. The method of claim 1 , wherein the at least one neoantigen-encoding nucleic acid sequence that is the same comprises an MHC class I epitope-encoding nucleic acid sequence.
13. The method of claim 1 , wherein the at least one neoantigene-encoding nucleic acid sequence that is the same comprises an epitope-encoding nucleic acid sequence comprising an alteration that makes the encoded epitope sequence distinct from a corresponding peptide sequence encoded by a wild-type nucleic acid sequence, optionally wherein the alteration is selected from the group consisting of: a point mutation, a frameshift mutation, a non-frameshift mutation, a deletion mutation, an insertion mutation, a splice variant, a genomic rearrangement, a proteasome-generated spliced neoantigen, and combinations thereof.
14. The method of claim 13 , wherein the encoded epitope comprising the alteration has (1) increased binding affinity to its corresponding MHC allele relative to the translated, corresponding wild-type nucleic acid sequence, (2) has increased binding stability to its corresponding MHC allele relative to the translated, corresponding wild-type nucleic acid sequence, and/or (3) has an increased likelihood of presentation on its corresponding MHC allele relative to the translated, corresponding wild-type nucleic acid sequence.
15. The method of claim 1 , wherein an ordered sequence of each element of one or both of the first neoantigen cassette and the second neoantigen cassette is described in the formula, from 5′ to 3′, comprising:
P a -( L 5 b - N c - L 3 d ) X -( G 5 e - U f ) Y - G 3 g
wherein P comprises a second promoter nucleotide sequence, where a=0 or 1,
N comprises an epitope-encoding nucleic acid sequences, where c=1,
L5 comprises a 5′ linker sequence, where b=0 or 1,
L3 comprises a 3′ linker sequence, where d=0 or 1,
G5 comprises at least one nucleic acid sequence encoding a GPGPG amino acid linker sequence (SEQ ID NO:56), where e=0 or 1,
G3 comprises at least one second nucleic acid sequence encoding a GPGPG amino acid linker sequence (SEQ ID NO:56), where g=0 or 1,
U comprises at least one MHC class II neoantigen-encoding nucleic acid sequence, where f=1,
X=2 to 400, and
Y=0, 1, or 2; optionally
(i) wherein for each X the corresponding N is distinct; and/or
(ii) wherein for each Y the corresponding U is distinct.
16. The method of claim 15 , wherein:
a=0, b=1, d=1, e=1, g=1, X≥2, Y≥0,
the at least one promoter nucleotide sequence is a single 26S promoter nucleotide sequence provided by the RNA alphavirus backbone,
the at least one polyadenylation poly(A) sequence is a poly(A) sequence of at least 80 consecutive A nucleotides provided by the RNA alphavirus backbone,
the epitope encoded by the epitope-encoding nucleic acid sequence comprises an MHC class I epitope, wherein the MHC class I epitope encoded by each N is 7-15 amino acids in length,
U is each of a PADRE class II sequence and a Tetanus toxoid MHC class II sequence,
the RNA alphavirus backbone is:
(a) the sequence of SEQ ID NO:3 further comprising a deletion between base pair 7544 and 11175, wherein the second neoantigen cassette is inserted to replace the deletion between base pair 7544 and 11175, and/or
(b) the sequence set forth in SEQ ID NO:6, wherein the second neoantigen cassette is inserted at position 7544 of the sequence set forth in SEQ ID NO:6 the sequence set forth in SEQ ID NO:6, and
each of the MHC class I neoantigen-encoding nucleic acid sequences encodes a polypeptide that is between 8 and 35 amino acids in length.
17. The method of claim 1 , wherein the at least one neoantigen-encoding nucleic acid sequence that is the same comprises an epitope-encoding nucleic acid sequence that encodes an epitope that is capable of being presented by an MHC allele on the surface of a cell of the subject, optionally wherein the cell is a tumor cell optionally selected from the group consisting of: lung cancer, melanoma, breast cancer, ovarian cancer, prostate cancer, kidney cancer, gastric cancer, colon cancer, testicular cancer, head and neck cancer, pancreatic cancer, bladder cancer, brain cancer, B-cell lymphoma, acute myelogenous leukemia, adult acute lymphoblastic leukemia, chronic myelogenous leukemia, chronic lymphocytic leukemia, T cell lymphocytic leukemia, non-small cell lung cancer, and small cell lung cancer.
18. The method of claim 9 , wherein the at least two neoantigen-encoding nucleic acid sequence comprises at least 2-10, 11-20, 15-20, 11-100, 11-200, 11-300, 11-400, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 2, 3, 4, 5, 6, 7, 8, 9, 10 nucleic acid sequences, or up to 400 nucleic acid sequences, optionally wherein at least two of the neoantigen-encoding nucleic acid sequences comprise an epitope-encoding nucleic acid sequence that encodes epitopes that are capable of being presented by an MHC allele on the surface of a cell of the subject.
19. The method of claim 1 , wherein the method further comprises administering (1) an adjuvant, and/or (2) an immune modulator, optionally wherein the immune modulator is an anti-CTLA4 antibody or an antigen-binding fragment thereof, an anti-PD-1 antibody or an antigen-binding fragment thereof, an anti-PD-L1 antibody or an antigen-binding fragment thereof, an anti-4-1BB antibody or an antigen-binding fragment thereof, or an anti-OX-40 antibody or an antigen-binding fragment thereof.
20. The method of claim 1 , wherein the method comprises treating a subject known or suspected of having cancer, and wherein the method further comprises obtaining or having obtained at least one of exome, transcriptome, or whole genome nucleotide sequencing data from the subject, optionally exome, transcriptome, or whole genome tumor nucleotide sequencing data from a tumor of the subject.