Compositions and methods for optimal peptide vaccines by residue optimization

Abstract

Immunogenic compositions are described that comprise a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOS: 19-50. Immunogenic compositions are also described that comprise a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOS: 141-272. Immunogenic compositions are also described that comprise a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOS: 273-322. Immunogenic compositions are also described that comprise a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOS: 354-458.

Description

[Technical field]

[0001] This application claims the benefit of and priority under 35 USC § 119(e) to U.S. patent application Ser. No. 17 / 243,096, filed April 28, 2021, the entire contents of which are incorporated herein by reference.

[0002] This patent disclosure contains material that is subject to copyright protection. The copyright holder has no objection to the facsimile reproduction of the patent document or patent disclosure as it appears in the U.S. Patent and Trademark Office patent file or records, but otherwise reserves all copyright rights.

[0003] Incorporation by Reference All patents, patent applications, and publications cited herein are hereby incorporated by reference in their entirety. All publications disclosed herein are hereby incorporated by reference in their entirety into this application.

[0004] Sequence Listing The Sequence Listing shall be submitted on one compact disc (Copy 1) and two copies thereof (Copy 2 and 3). Each disc contains one file, each entitled "2215269_00125US1_SL.txt", in ASCII format, created on April 26, 2021. Each compact disc has been created in Macintosh machine format and is compatible with the Macintosh operating system. The material contained on the compact discs is specifically incorporated herein by reference.

[0005] Technical Field The present invention relates generally to peptide vaccine compositions, systems, and methods. More specifically, the present invention relates to compositions, systems, and methods for designing peptide vaccines that treat or prevent disease, optimized based on predicted population immunogenicity. [Background technology]

[0006] The goal of peptide vaccines is to train the immune system to expand its ability to recognize and bind to cells that present the target peptide, improving the immune response against cancer cells or pathogens. Peptide vaccines can also be administered to individuals already suffering from disease to boost the immune response against the underlying cancer, other disease, or pathogen. Alternatively, peptide vaccines can be administered to provide the immune system with therapeutic resistance to one or more peptides. There is a need for compositions, systems, and methods of peptide vaccines based on target peptides that are predicted to be presented to protect the host against cancer, other diseases, or pathogen infection. Summary of the Invention

[0007] In one aspect, the invention provides a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 1 to 474.

[0008] In some embodiments, the nucleic acid sequence encodes two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 1-474.

[0009] In another aspect, the present invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 1 to 474.

[0010] In some embodiments, an immunogenic composition is administered to a subject. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOS: 1 to 474. In some embodiments, the nucleic acid sequence is administered as a construct for expression in vivo. In some embodiments, the in vivo administration of the nucleic acid sequence is configured to produce one or more peptides that are presented by HLA class I molecules. In some embodiments, the one or more peptides are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, TP53. In some embodiments, the one or more peptides are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 The immunogenic composition is a modified or unmodified fragment of a protein comprising a mutation selected from the group consisting of TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the immunogenic composition is administered to a subject in an effective amount to prevent cancer. In some embodiments, the immunogenic composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the cancer is selected from the group consisting of pancreatic cancer, skin cancer, thyroid cancer, brain cancer, colorectal cancer, bronchial and lung cancer, breast cancer, and ovarian cancer.In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding at least three amino acid sequences selected from the group consisting of SEQ ID NOs: 1 to 474.

[0011] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 1 to 474.

[0012] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 1-474.

[0013] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 1 to 18.

[0014] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from an AKT1 protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 1 to 18.

[0015] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 1-18.

[0016] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 1 to 18. In some embodiments, one or more of the peptides is a modified or unmodified fragment of a mutated AKT1 protein.

[0017] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 19 to 50.

[0018] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a BRAF protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 19 to 50.

[0019] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 19 to 50.

[0020] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 19 to 50. In some embodiments, one or more of the peptides is a modified or unmodified fragment of a mutated BRAF protein.

[0021] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 51 to 98.

[0022] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from an EGFR protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 51 to 98.

[0023] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 51 to 98.

[0024] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 51 to 98. In some embodiments, one or more of the peptides is a modified or unmodified fragment of a mutated EGFR protein.

[0025] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 99 to 118.

[0026] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a GTF2I protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 99 to 118.

[0027] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 99 to 118.

[0028] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 99 to 118. In some embodiments, one or more of the peptides is a modified or unmodified fragment of a mutated GTF2I protein.

[0029] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 119 to 140.

[0030] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from an IDH1 protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 119 to 140.

[0031] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 119 to 140.

[0032] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 119 to 140. In some embodiments, one or more of the peptides is a modified or unmodified fragment of a mutated IDH1 protein.

[0033] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 141 to 229.

[0034] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a KRAS protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 141 to 229.

[0035] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 141 to 229.

[0036] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 141 to 229. In some embodiments, one or more of the peptides is a modified or unmodified fragment of a mutated KRAS protein.

[0037] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 230 to 272.

[0038] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a NRAS protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 230 to 272.

[0039] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 230 to 272.

[0040] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 230 to 272. In some embodiments, one or more of the peptides is a modified or unmodified fragment of a mutated NRAS protein.

[0041] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 273 to 322.

[0042] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a PIK3CA protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 273 to 322.

[0043] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 273 to 322.

[0044] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 273 to 322. In some embodiments, one or more of the peptides is a modified or unmodified fragment of a mutated PIK3CA protein.

[0045] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 323 to 353.

[0046] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a PTEN protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 323 to 353.

[0047] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 323 to 353.

[0048] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 323 to 353. In some embodiments, one or more of the peptides is a modified or unmodified fragment of a mutated PTEN protein.

[0049] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 354 to 458.

[0050] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a TP53 protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 354 to 458.

[0051] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 354 to 458.

[0052] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 354 to 458. In some embodiments, one or more of the peptides is a modified or unmodified fragment of a mutated TP53 protein.

[0053] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 141 to 272.

[0054] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a RAS protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 141 to 272.

[0055] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 141 to 272.

[0056] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 141 to 272. In some embodiments, one or more of the peptides is a modified or unmodified fragment of a mutated RAS protein.

[0057] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 19 to 33.

[0058] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a BRAF V600E protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 19 to 33.

[0059] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 19 to 33.

[0060] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 19 to 33. In some embodiments, the immunogenic peptide composition comprises peptides derived from a BRAF V600E protein mutation.

[0061] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 34 to 50.

[0062] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a BRAF V600M protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 34 to 50.

[0063] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 34 to 50.

[0064] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 34 to 50. In some embodiments, the immunogenic peptide composition comprises peptides derived from a BRAF V600M protein mutation.

[0065] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 51 to 66.

[0066] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from an EGFR A289V protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 51 to 66.

[0067] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 51 to 66.

[0068] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 51 to 66. In some embodiments, the immunogenic peptide composition comprises peptides derived from the EGFR A289V protein mutation.

[0069] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 67 to 81.

[0070] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from an EGFR G598V protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 67 to 81.

[0071] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 67 to 81.

[0072] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 67 to 81. In some embodiments, the immunogenic peptide composition comprises a peptide derived from the EGFR G598V protein mutation.

[0073] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 82 to 98.

[0074] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from an EGFR L858R protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 82 to 98.

[0075] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 82 to 98.

[0076] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 82 to 98. In some embodiments, the immunogenic peptide composition comprises peptides derived from the EGFR L858R protein mutation.

[0077] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 125 to 140.

[0078] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from an IDH1 R132H protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 125 to 140.

[0079] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 125 to 140.

[0080] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 125 to 140. In some embodiments, the immunogenic peptide composition comprises peptides derived from an IDH1 R132H protein mutation.

[0081] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 119 to 124.

[0082] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from an IDH1 R132C protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 119 to 124.

[0083] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 119 to 124.

[0084] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 119 to 124. In some embodiments, the immunogenic peptide composition comprises peptides derived from the IDH1 R132C protein mutation.

[0085] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 167 to 178.

[0086] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a KRAS G12D protein mutation, in some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 167 to 178.

[0087] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 167 to 178.

[0088] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 167 to 178. In some embodiments, the immunogenic peptide composition comprises peptides derived from a KRAS G12D protein mutation.

[0089] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 203 to 213.

[0090] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a KRAS G12V protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 203 to 213.

[0091] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 203 to 213.

[0092] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 203 to 213. In some embodiments, the immunogenic peptide composition comprises peptides derived from a KRAS G12V protein mutation.

[0093] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 179 to 191.

[0094] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a KRAS G12R protein mutation, in some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 179 to 191.

[0095] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 179 to 191.

[0096] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 179 to 191. In some embodiments, the immunogenic peptide composition comprises peptides derived from a KRAS G12R protein mutation.

[0097] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 154 to 166.

[0098] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a KRAS G12C protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 154 to 166.

[0099] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 154 to 166.

[0100] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 154 to 166. In some embodiments, the immunogenic peptide composition comprises peptides derived from a KRAS G12C protein mutation.

[0101] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 214 to 229.

[0102] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a KRAS G13D protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 214 to 229.

[0103] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 214 to 229.

[0104] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 214 to 229. In some embodiments, the immunogenic peptide composition comprises peptides derived from a KRAS G13D protein mutation.

[0105] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 141 to 153.

[0106] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a KRAS G12A protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 141 to 153.

[0107] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 141 to 153.

[0108] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 141 to 153. In some embodiments, the immunogenic peptide composition comprises peptides derived from a KRAS G12A protein mutation.

[0109] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 192 to 202.

[0110] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a KRAS G12S protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 192 to 202.

[0111] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 192 to 202.

[0112] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 192 to 202. In some embodiments, the immunogenic peptide composition comprises peptides derived from a KRAS G12S protein mutation.

[0113] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 256 to 272.

[0114] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from the NRAS Q61R protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 256 to 272.

[0115] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 256 to 272.

[0116] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 256 to 272. In some embodiments, the immunogenic peptide composition comprises peptides derived from the NRAS Q61R protein mutation.

[0117] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 230 to 238.

[0118] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from the NRAS Q61K protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 230 to 238.

[0119] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 230 to 238.

[0120] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 230 to 238. In some embodiments, the immunogenic peptide composition comprises peptides derived from the NRAS Q61K protein mutation.

[0121] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 239 to 255.

[0122] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from an NRAS Q61L protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 239 to 255.

[0123] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 239 to 255.

[0124] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 239 to 255. In some embodiments, the immunogenic peptide composition comprises peptides derived from the NRAS Q61L protein mutation.

[0125] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 273 to 285.

[0126] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a PIK3CA E542K protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 273 to 285.

[0127] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 273 to 285.

[0128] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 273 to 285. In some embodiments, the immunogenic peptide composition comprises peptides derived from the PIK3CA E542K protein mutation.

[0129] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 286 to 293.

[0130] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a PIK3CA E545K protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 286 to 293.

[0131] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 286 to 293.

[0132] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 286 to 293. In some embodiments, the immunogenic peptide composition comprises peptides derived from a PIK3CA E545K protein mutation.

[0133] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOS: 294 to 309.

[0134] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a PIK3CA H1047R protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 294 to 309.

[0135] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 294 to 309.

[0136] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 294 to 309. In some embodiments, the immunogenic peptide composition comprises peptides derived from the PIK3CA H1047R protein mutation.

[0137] In another aspect, the present invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 359 to 374.

[0138] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a TP53 R158L protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 359 to 374.

[0139] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 359 to 374.

[0140] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 359 to 374. In some embodiments, the immunogenic peptide composition comprises peptides derived from the TP53 R158L protein mutation.

[0141] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 375 to 386.

[0142] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a TP53 R175H protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 375 to 386.

[0143] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 375 to 386.

[0144] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 375 to 386. In some embodiments, the immunogenic peptide composition comprises peptides derived from the TP53 R175H protein mutation.

[0145] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOS: 387 to 401.

[0146] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a TP53 R248Q protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 387 to 401.

[0147] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOS: 387 to 401.

[0148] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 387 to 401. In some embodiments, the immunogenic peptide composition comprises peptides derived from the TP53 R248Q protein mutation.

[0149] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 422 to 432.

[0150] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from the TP53 R273C protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 422 to 432.

[0151] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 422 to 432.

[0152] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 422 to 432. In some embodiments, the immunogenic peptide composition comprises peptides derived from the TP53 R273C protein mutation.

[0153] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 433 to 446.

[0154] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from the TP53 R273H protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 433 to 446.

[0155] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 433 to 446.

[0156] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 433 to 446. In some embodiments, the immunogenic peptide composition comprises peptides derived from the TP53 R273H protein mutation.

[0157] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 402 to 421.

[0158] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from the TP53 R248W protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 402 to 421.

[0159] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 402 to 421.

[0160] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 402 to 421. In some embodiments, the immunogenic peptide composition comprises peptides derived from the TP53 R248W protein mutation.

[0161] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 447 to 449.

[0162] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from the TP53 R282W protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 447 to 449.

[0163] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 447 to 449.

[0164] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 447 to 449. In some embodiments, the immunogenic peptide composition comprises peptides derived from the TP53 R282W protein mutation.

[0165] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 450 to 458.

[0166] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a TP53 Y220C protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 450 to 458.

[0167] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 450 to 458.

[0168] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 450 to 458. In some embodiments, the immunogenic peptide composition comprises peptides derived from the TP53 Y220C protein mutation.

[0169] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 310 to 322.

[0170] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a PIK3CA R88Q protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 310 to 322.

[0171] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 310 to 322.

[0172] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 310 to 322. In some embodiments, the immunogenic peptide composition comprises peptides derived from a PIK3CA R88Q protein mutation.

[0173] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 99 to 118.

[0174] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a GTF2I L424H protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 99 to 118.

[0175] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 99 to 118.

[0176] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 99 to 118. In some embodiments, the immunogenic peptide composition comprises peptides derived from the GTF2I L424H protein mutation.

[0177] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 338 to 353.

[0178] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a PTEN R130Q protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 338 to 353.

[0179] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 338 to 353.

[0180] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 338 to 353. In some embodiments, the immunogenic peptide composition comprises peptides derived from the PTEN R130Q protein mutation.

[0181] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 1 to 18.

[0182] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from an AKT1 E17K protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 1 to 18.

[0183] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 1-18.

[0184] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 1 to 18. In some embodiments, the immunogenic peptide composition comprises a peptide derived from the AKT1 E17K protein mutation.

[0185] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 323 to 337.

[0186] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a PTEN R130G protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 323 to 337.

[0187] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 323 to 337.

[0188] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 323 to 337. In some embodiments, the immunogenic peptide composition comprises peptides derived from the PTEN R130G protein mutation.

[0189] In another aspect, the present invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 354 to 358.

[0190] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from the TP53 H179R protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 354 to 358.

[0191] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 354 to 358.

[0192] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 354 to 358. In some embodiments, the immunogenic peptide composition comprises peptides derived from the TP53 H179R protein mutation.

[0193] In another aspect, the present invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 168 to 169, SEQ ID NO: 171, SEQ ID NOs: 179 to 180, SEQ ID NOs: 182 to 183, SEQ ID NOs: 203 to 204, and SEQ ID NO: 207.

[0194] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 168 to 169, SEQ ID NO: 171, SEQ ID NOs: 179 to 180, SEQ ID NOs: 182 to 183, SEQ ID NOs: 203 to 204, and SEQ ID NO: 207. In some embodiments, the nucleic acid sequence is administered as a construct for expression in vivo. In some embodiments, the administration of the nucleic acid sequence in vivo is configured to produce one or more peptides that are presented by HLA class I molecules. In some embodiments, the one or more peptides are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, and TP53. In some embodiments, the one or more peptides are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 The nucleic acid sequence is a modified or unmodified fragment of a protein having a mutation selected from the group consisting of TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the nucleic acid sequence is administered to a subject in an effective amount to prevent cancer. In some embodiments, the nucleic acid sequence is administered to a subject in an effective amount to treat cancer. In some embodiments, the cancer is pancreatic cancer.

[0195] In another aspect, the present invention provides a method of treating or preventing pancreatic cancer by administering to a subject an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 168 to 169, SEQ ID NO: 171, SEQ ID NOs: 179 to 180, SEQ ID NOs: 182 to 183, SEQ ID NOs: 203 to 204, and SEQ ID NO: 207.

[0196] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 168 to 169, SEQ ID NO: 171, SEQ ID NOs: 179 to 180, SEQ ID NOs: 182 to 183, SEQ ID NOs: 203 to 204, and SEQ ID NO: 207.

[0197] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 168 to 169, SEQ ID NO: 171, SEQ ID NOs: 179 to 180, SEQ ID NOs: 182 to 183, SEQ ID NOs: 203 to 204, and SEQ ID NO: 207.

[0198] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 168 to 169, SEQ ID NO: 171, SEQ ID NOs: 179 to 180, SEQ ID NOs: 182 to 183, SEQ ID NOs: 203 to 204, SEQ ID NO: 207. In some embodiments, the peptides in the immunogenic peptide composition are presented by HLA class I molecules. In some embodiments, the peptides in the immunogenic peptide composition are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, TP53. In some embodiments, the peptides in the immunogenic peptide composition are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 The modified or unmodified fragment of the protein comprising a mutation selected from the group consisting of TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the immunogenic peptide composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the immunogenic peptide composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the cancer is pancreatic cancer.

[0199] In another aspect, the present invention provides a method of treating or preventing pancreatic cancer by administering to a subject an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 168 to 169, SEQ ID NO: 171, SEQ ID NOs: 179 to 180, SEQ ID NOs: 182 to 183, SEQ ID NOs: 203 to 204, and SEQ ID NO: 207.

[0200] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 168 to 169, SEQ ID NO: 171, SEQ ID NOs: 179 to 180, SEQ ID NOs: 182 to 183, SEQ ID NOs: 203 to 204, and SEQ ID NO: 207.

[0201] In another aspect, the present invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 19 to 23, SEQ ID NOs: 34 to 40, SEQ ID NOs: 230 to 231, SEQ ID NOs: 239, SEQ ID NOs: 241 to 242, and SEQ ID NOs: 260 to 262.

[0202] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 19 to 23, SEQ ID NOs: 34 to 40, SEQ ID NOs: 230 to 231, SEQ ID NOs: 239, SEQ ID NOs: 241 to 242, and SEQ ID NOs: 260 to 262. In some embodiments, the nucleic acid sequence is administered as a construct for expression in vivo. In some embodiments, the in vivo administration of the nucleic acid sequence is configured to produce one or more peptides that are presented by HLA class I molecules. In some embodiments, the one or more peptides are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, and TP53. In some embodiments, the one or more peptides are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 The nucleic acid sequence is a modified or unmodified fragment of a protein having a mutation selected from the group consisting of TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the nucleic acid sequence is administered to a subject in an effective amount to prevent cancer. In some embodiments, the nucleic acid sequence is administered to a subject in an effective amount to treat cancer. In some embodiments, the cancer is skin cancer.

[0203] In another aspect, the present invention provides a method of treating or preventing skin cancer by administering to a subject an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 19 to 23, SEQ ID NOs: 34 to 40, SEQ ID NOs: 230 to 231, SEQ ID NO: 239, SEQ ID NOs: 241 to 242, and SEQ ID NOs: 260 to 262.

[0204] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 19 to 23, SEQ ID NOs: 34 to 40, SEQ ID NOs: 230 to 231, SEQ ID NOs: 239, SEQ ID NOs: 241 to 242, and SEQ ID NOs: 260 to 262.

[0205] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 19 to 23, SEQ ID NOs: 34 to 40, SEQ ID NOs: 230 to 231, SEQ ID NOs: 239, SEQ ID NOs: 241 to 242, and SEQ ID NOs: 260 to 262.

[0206] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 19 to 23, SEQ ID NOs: 34 to 40, SEQ ID NOs: 230 to 231, SEQ ID NOs: 239, SEQ ID NOs: 241 to 242, SEQ ID NOs: 260 to 262. In some embodiments, the peptides in the immunogenic peptide composition are presented by HLA class I molecules. In some embodiments, the peptides in the immunogenic peptide composition are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, TP53. In some embodiments, the peptides in the immunogenic peptide composition are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 The modified or unmodified fragment of the protein comprising a mutation selected from the group consisting of TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the immunogenic peptide composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the immunogenic peptide composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the cancer is skin cancer.

[0207] In another aspect, the present invention provides a method of treating or preventing skin cancer by administering to a subject an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 19 to 23, SEQ ID NOs: 34 to 40, SEQ ID NOs: 230 to 231, SEQ ID NO: 239, SEQ ID NOs: 241 to 242, and SEQ ID NOs: 260 to 262.

[0208] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 19 to 23, SEQ ID NOs: 34 to 40, SEQ ID NOs: 230 to 231, SEQ ID NO: 239, SEQ ID NOs: 241 to 242, and SEQ ID NOs: 260 to 262.

[0209] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 19 to 23, SEQ ID NOs: 230 to 231, and SEQ ID NOs: 260 to 262.

[0210] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOS: 19 to 23, SEQ ID NOS: 230 to 231, and SEQ ID NOS: 260 to 262. In some embodiments, the nucleic acid sequence is administered as a construct for expression in vivo. In some embodiments, the in vivo administration of the nucleic acid sequence is configured to produce one or more peptides that are presented by an HLA class I molecule. In some embodiments, the one or more peptides are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, and TP53. In some embodiments, the one or more peptides are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 The nucleic acid sequence is a modified or unmodified fragment of a protein having a mutation selected from the group consisting of TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the nucleic acid sequence is administered to a subject in an effective amount to prevent cancer. In some embodiments, the nucleic acid sequence is administered to a subject in an effective amount to treat cancer. In some embodiments, the cancer is thyroid cancer.

[0211] In another aspect, the invention provides a method of treating or preventing thyroid cancer by administering to a subject an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 19 to 23, SEQ ID NOs: 230 to 231, and SEQ ID NOs: 260 to 262.

[0212] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 19 to 23, SEQ ID NOs: 230 to 231, and SEQ ID NOs: 260 to 262.

[0213] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 19 to 23, SEQ ID NOs: 230 to 231, and SEQ ID NOs: 260 to 262.

[0214] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 19 to 23, SEQ ID NOs: 230 to 231, and SEQ ID NOs: 260 to 262. In some embodiments, the peptides in the immunogenic peptide composition are presented by HLA class I molecules. In some embodiments, the peptides in the immunogenic peptide composition are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, and TP53. In some embodiments, the peptides in the immunogenic peptide composition are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 The modified or unmodified fragment of the protein comprising a mutation selected from the group consisting of TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the immunogenic peptide composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the immunogenic peptide composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the cancer is thyroid cancer.

[0215] In another aspect, the invention provides a method of treating or preventing thyroid cancer by administering to a subject an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 19 to 23, SEQ ID NOs: 230 to 231, and SEQ ID NOs: 260 to 262.

[0216] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 19 to 23, SEQ ID NOs: 230 to 231, and SEQ ID NOs: 260 to 262.

[0217] In another aspect, the present invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 51 to 55, SEQ ID NOs: 67 to 70, SEQ ID NOs: 72 to 73, SEQ ID NOs: 119 to 120, SEQ ID NOs: 125 to 130, SEQ ID NO: 375, SEQ ID NO: 423, and SEQ ID NO: 425.

[0218] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 51 to 55, SEQ ID NOs: 67 to 70, SEQ ID NOs: 72 to 73, SEQ ID NOs: 119 to 120, SEQ ID NOs: 125 to 130, SEQ ID NO: 375, SEQ ID NO: 423, SEQ ID NO: 425. In some embodiments, the nucleic acid sequence is administered as a construct for expression in vivo. In some embodiments, the in vivo administration of the nucleic acid sequence is configured to produce one or more peptides that are presented by HLA class I molecules. In some embodiments, the one or more peptides are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, TP53. In some embodiments, the one or more peptides are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 The nucleic acid sequence is a modified or unmodified fragment of a protein having a mutation selected from the group consisting of TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the nucleic acid sequence is administered to a subject in an effective amount to prevent cancer. In some embodiments, the nucleic acid sequence is administered to a subject in an effective amount to treat cancer. In some embodiments, the cancer is a brain tumor.

[0219] In another aspect, the present invention provides a method of treating or preventing brain tumors by administering to a subject an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 51 to 55, SEQ ID NOs: 67 to 70, SEQ ID NOs: 72 to 73, SEQ ID NOs: 119 to 120, SEQ ID NOs: 125 to 130, SEQ ID NO: 375, SEQ ID NO: 423, and SEQ ID NO: 425.

[0220] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 51 to 55, SEQ ID NOs: 67 to 70, SEQ ID NOs: 72 to 73, SEQ ID NOs: 119 to 120, SEQ ID NOs: 125 to 130, SEQ ID NO: 375, SEQ ID NO: 423, and SEQ ID NO: 425.

[0221] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 51 to 55, SEQ ID NOs: 67 to 70, SEQ ID NOs: 72 to 73, SEQ ID NOs: 119 to 120, SEQ ID NOs: 125 to 130, SEQ ID NO: 375, SEQ ID NO: 423, and SEQ ID NO: 425.

[0222] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 51 to 55, SEQ ID NOs: 67 to 70, SEQ ID NOs: 72 to 73, SEQ ID NOs: 119 to 120, SEQ ID NOs: 125 to 130, SEQ ID NO: 375, SEQ ID NO: 423, SEQ ID NO: 425. In some embodiments, the peptides in the immunogenic peptide composition are presented by HLA class I molecules. In some embodiments, the peptides in the immunogenic peptide composition are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, TP53. In some embodiments, the peptides in the immunogenic peptide composition are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 The modified or unmodified fragment of the protein comprising a mutation selected from the group consisting of TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the immunogenic peptide composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the immunogenic peptide composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the cancer is a brain tumor.

[0223] In another aspect, the present invention provides a method of treating or preventing brain tumors by administering to a subject an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 51 to 55, SEQ ID NOs: 67 to 70, SEQ ID NOs: 72 to 73, SEQ ID NOs: 119 to 120, SEQ ID NOs: 125 to 130, SEQ ID NO: 375, SEQ ID NO: 423, and SEQ ID NO: 425.

[0224] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 51 to 55, SEQ ID NOs: 67 to 70, SEQ ID NOs: 72 to 73, SEQ ID NOs: 119 to 120, SEQ ID NOs: 125 to 130, SEQ ID NO: 375, SEQ ID NO: 423, and SEQ ID NO: 425.

[0225] In another aspect, the present invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 20 to 23, SEQ ID NOs: 167 to 169, SEQ ID NO: 171, SEQ ID NOs: 203 to 207, SEQ ID NOs: 215 to 217, and SEQ ID NO: 288.

[0226] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 20 to 23, SEQ ID NOs: 167 to 169, SEQ ID NO: 171, SEQ ID NOs: 203 to 207, SEQ ID NOs: 215 to 217, and SEQ ID NO: 288. In some embodiments, the nucleic acid sequence is administered as a construct for expression in vivo. In some embodiments, the administration of the nucleic acid sequence in vivo is configured to produce one or more peptides that are presented by HLA class I molecules. In some embodiments, the one or more peptides are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, and TP53. In some embodiments, the one or more peptides are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 The nucleic acid sequence is a modified or unmodified fragment of a protein having a mutation selected from the group consisting of TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the nucleic acid sequence is administered to a subject in an effective amount to prevent cancer. In some embodiments, the nucleic acid sequence is administered to a subject in an effective amount to treat cancer. In some embodiments, the cancer is colorectal cancer.

[0227] In another aspect, the present invention provides a method of treating or preventing colorectal cancer by administering to a subject an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 20 to 23, SEQ ID NOs: 167 to 169, SEQ ID NO: 171, SEQ ID NOs: 203 to 207, SEQ ID NOs: 215 to 217, and SEQ ID NO: 288.

[0228] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 20 to 23, SEQ ID NOs: 167 to 169, SEQ ID NO: 171, SEQ ID NOs: 203 to 207, SEQ ID NOs: 215 to 217, and SEQ ID NO: 288.

[0229] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 20 to 23, SEQ ID NOs: 167 to 169, SEQ ID NO: 171, SEQ ID NOs: 203 to 207, SEQ ID NOs: 215 to 217, and SEQ ID NO: 288.

[0230] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 20 to 23, SEQ ID NOs: 167 to 169, SEQ ID NO: 171, SEQ ID NOs: 203 to 207, SEQ ID NOs: 215 to 217, SEQ ID NO: 288. In some embodiments, the peptides in the immunogenic peptide composition are presented by HLA class I molecules. In some embodiments, the peptides in the immunogenic peptide composition are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, TP53. In some embodiments, the peptides in the immunogenic peptide composition are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 The modified or unmodified fragment of the protein comprising a mutation selected from the group consisting of TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the immunogenic peptide composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the immunogenic peptide composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the cancer is colorectal cancer.

[0231] In another aspect, the present invention provides a method of treating or preventing colorectal cancer by administering to a subject an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 20 to 23, SEQ ID NOs: 167 to 169, SEQ ID NO: 171, SEQ ID NOs: 203 to 207, SEQ ID NOs: 215 to 217, and SEQ ID NO: 288.

[0232] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 20 to 23, SEQ ID NOs: 167 to 169, SEQ ID NO: 171, SEQ ID NOs: 203 to 207, SEQ ID NOs: 215 to 217, and SEQ ID NO: 288.

[0233] In another aspect, the present invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs:82 to 86, SEQ ID NOs:141 to 144, SEQ ID NOs:154 to 159, SEQ ID NOs:168 to 169, SEQ ID NO:171, SEQ ID NOs:203 to 204, SEQ ID NO:207, SEQ ID NOs:274 to 276, SEQ ID NO:288, SEQ ID NO:359, and SEQ ID NOs:362 to 364.

[0234] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 82 to 86, SEQ ID NOs: 141 to 144, SEQ ID NOs: 154 to 159, SEQ ID NOs: 168 to 169, SEQ ID NO: 171, SEQ ID NOs: 203 to 204, SEQ ID NO: 207, SEQ ID NOs: 274 to 276, SEQ ID NO: 288, SEQ ID NO: 359, SEQ ID NOs: 362 to 364. In some embodiments, the nucleic acid sequence is administered as a construct for expression in vivo. In some embodiments, the in vivo administration of the nucleic acid sequence is configured to produce one or more peptides that are presented by an HLA class I molecule. In some embodiments, the one or more peptides are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, TP53. In some embodiments, the one or more peptides are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the nucleic acid sequence is administered to a subject in an effective amount to prevent cancer. In some embodiments, the nucleic acid sequence is administered to a subject in an effective amount to treat cancer.In some embodiments, the cancer is bronchial and lung cancer.

[0235] In another aspect, the present invention provides a method of treating or preventing bronchial and lung cancer by administering to a subject an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs:82 to 86, SEQ ID NOs:141 to 144, SEQ ID NOs:154 to 159, SEQ ID NOs:168 to 169, SEQ ID NO:171, SEQ ID NOs:203 to 204, SEQ ID NO:207, SEQ ID NOs:274 to 276, SEQ ID NO:288, SEQ ID NO:359, and SEQ ID NOs:362 to 364.

[0236] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 82 to 86, SEQ ID NOs: 141 to 144, SEQ ID NOs: 154 to 159, SEQ ID NOs: 168 to 169, SEQ ID NO: 171, SEQ ID NOs: 203 to 204, SEQ ID NO: 207, SEQ ID NOs: 274 to 276, SEQ ID NO: 288, SEQ ID NO: 359, and SEQ ID NOs: 362 to 364.

[0237] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 82 to 86, SEQ ID NOs: 141 to 144, SEQ ID NOs: 154 to 159, SEQ ID NOs: 168 to 169, SEQ ID NO: 171, SEQ ID NOs: 203 to 204, SEQ ID NO: 207, SEQ ID NOs: 274 to 276, SEQ ID NO: 288, SEQ ID NO: 359, and SEQ ID NOs: 362 to 364.

[0238] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 82-86, SEQ ID NOs: 141-144, SEQ ID NOs: 154-159, SEQ ID NOs: 168-169, SEQ ID NO: 171, SEQ ID NOs: 203-204, SEQ ID NO: 207, SEQ ID NOs: 274-276, SEQ ID NO: 288, SEQ ID NO: 359, SEQ ID NOs: 362-364. In some embodiments, the peptides in the immunogenic peptide composition are presented by HLA class I molecules. In some embodiments, the peptides in the immunogenic peptide composition are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, TP53. In some embodiments, the peptides in the immunogenic peptide composition are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 The modified or unmodified fragment of the protein comprising a mutation selected from the group consisting of TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the immunogenic peptide composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the immunogenic peptide composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the cancer is bronchial cancer and lung cancer.

[0239] In another aspect, the present invention provides a method of treating or preventing bronchial and lung cancer by administering to a subject an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs:82 to 86, SEQ ID NOs:141 to 144, SEQ ID NOs:154 to 159, SEQ ID NOs:168 to 169, SEQ ID NO:171, SEQ ID NOs:203 to 204, SEQ ID NO:207, SEQ ID NOs:274 to 276, SEQ ID NO:288, SEQ ID NO:359, and SEQ ID NOs:362 to 364.

[0240] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 82 to 86, SEQ ID NOs: 141 to 144, SEQ ID NOs: 154 to 159, SEQ ID NOs: 168 to 169, SEQ ID NO: 171, SEQ ID NOs: 203 to 204, SEQ ID NO: 207, SEQ ID NOs: 274 to 276, SEQ ID NO: 288, SEQ ID NO: 359, and SEQ ID NOs: 362 to 364.

[0241] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 273 to 309, SEQ ID NOs: 375 to 386, and SEQ ID NOs: 422 to 446.

[0242] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOS: 273 to 309, 375 to 386, and 422 to 446. In some embodiments, the nucleic acid sequence is administered as a construct for expression in vivo. In some embodiments, the in vivo administration of the nucleic acid sequence is configured to produce one or more peptides that are presented by an HLA class I molecule. In some embodiments, the one or more peptides are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, and TP53. In some embodiments, the one or more peptides are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 The nucleic acid sequence is a modified or unmodified fragment of a protein having a mutation selected from the group consisting of TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the nucleic acid sequence is administered to a subject in an effective amount to prevent cancer. In some embodiments, the nucleic acid sequence is administered to a subject in an effective amount to treat cancer. In some embodiments, the cancer is breast cancer.

[0243] In another aspect, the invention provides a method of treating or preventing breast cancer by administering to a subject an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOS: 273 to 309, 375 to 386, and 422 to 446.

[0244] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 273 to 309, SEQ ID NOs: 375 to 386, and SEQ ID NOs: 422 to 446.

[0245] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 273 to 309, 375 to 386, and 422 to 446.

[0246] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOS: 273 to 309, 375 to 386, and 422 to 446. In some embodiments, the peptides in the immunogenic peptide composition are presented by HLA class I molecules. In some embodiments, the peptides in the immunogenic peptide composition are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, and TP53. In some embodiments, the peptides in the immunogenic peptide composition are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 The modified or unmodified fragment of the protein comprising a mutation selected from the group consisting of TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the immunogenic peptide composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the immunogenic peptide composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the cancer is breast cancer.

[0247] In another aspect, the invention provides a method of treating or preventing breast cancer by administering to a subject an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOS: 273 to 309, 375 to 386, and 422 to 446.

[0248] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 273 to 309, SEQ ID NOs: 375 to 386, and SEQ ID NOs: 422 to 446.

[0249] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 273 to 309, SEQ ID NOs: 375 to 386, and SEQ ID NOs: 422 to 446.

[0250] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOS: 273 to 309, 375 to 386, and 422 to 446. In some embodiments, the nucleic acid sequence is administered as a construct for expression in vivo. In some embodiments, the in vivo administration of the nucleic acid sequence is configured to produce one or more peptides that are presented by an HLA class I molecule. In some embodiments, the one or more peptides are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, and TP53. In some embodiments, the one or more peptides are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 The nucleic acid sequence is a modified or unmodified fragment of a protein having a mutation selected from the group consisting of TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the nucleic acid sequence is administered to a subject in an effective amount to prevent cancer. In some embodiments, the nucleic acid sequence is administered to a subject in an effective amount to treat cancer. In some embodiments, the cancer is ovarian cancer.

[0251] In another aspect, the present invention provides a method of treating or preventing ovarian cancer by administering to a subject an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOS: 273 to 309, 375 to 386, and 422 to 446.

[0252] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 273 to 309, SEQ ID NOs: 375 to 386, and SEQ ID NOs: 422 to 446.

[0253] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 273 to 309, 375 to 386, and 422 to 446.

[0254] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOS: 273 to 309, 375 to 386, and 422 to 446. In some embodiments, the peptides in the immunogenic peptide composition are presented by HLA class I molecules. In some embodiments, the peptides in the immunogenic peptide composition are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, and TP53. In some embodiments, the peptides in the immunogenic peptide composition are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 The immunogenic peptide composition is a modified or unmodified fragment of a protein comprising a mutation selected from the group consisting of TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the immunogenic peptide composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the immunogenic peptide composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the cancer is ovarian cancer.

[0255] In another aspect, the present invention provides a method of treating or preventing ovarian cancer by administering to a subject an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOS: 273 to 309, 375 to 386, and 422 to 446.

[0256] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 273 to 309, SEQ ID NOs: 375 to 386, and SEQ ID NOs: 422 to 446.

[0257] In one aspect, the invention provides a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 475 to 759.

[0258] In some embodiments, the nucleic acid sequence encodes two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 475 to 759.

[0259] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 475 to 759.

[0260] In some embodiments, an immunogenic composition is administered to a subject. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOS: 475 to 759. In some embodiments, the nucleic acid sequence is administered as a construct for expression in vivo. In some embodiments, the in vivo administration of the nucleic acid sequence is configured to produce one or more peptides that are presented by HLA class II molecules. In some embodiments, the one or more peptides are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, TP53. In some embodiments, the one or more peptides are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 The immunogenic composition is a modified or unmodified fragment of a protein comprising a mutation selected from the group consisting of TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the immunogenic composition is administered to a subject in an effective amount to prevent cancer. In some embodiments, the immunogenic composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the cancer is selected from the group consisting of pancreatic cancer, skin cancer, thyroid cancer, brain cancer, colorectal cancer, bronchial and lung cancer, breast cancer, and ovarian cancer.In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding at least three amino acid sequences selected from the group consisting of SEQ ID NOs: 475 to 759.

[0261] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 475 to 759.

[0262] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 475 to 759.

[0263] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 475 to 483.

[0264] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from an AKT1 protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 475 to 483.

[0265] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 475 to 483.

[0266] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 475 to 483. In some embodiments, one or more of the peptides is a modified or unmodified fragment of a mutated AKT1 protein.

[0267] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 484 to 502.

[0268] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a BRAF protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 484 to 502.

[0269] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 484 to 502.

[0270] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 484 to 502. In some embodiments, one or more of the peptides is a modified or unmodified fragment of a mutated BRAF protein.

[0271] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 503 to 527.

[0272] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from an EGFR protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 503 to 527.

[0273] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 503 to 527.

[0274] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 503 to 527. In some embodiments, one or more of the peptides is a modified or unmodified fragment of a mutated EGFR protein.

[0275] In another aspect, the present invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 528 to 534.

[0276] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a GTF2I protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 528 to 534.

[0277] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 528 to 534.

[0278] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 528 to 534. In some embodiments, one or more of the peptides is a modified or unmodified fragment of a mutated GTF2I protein.

[0279] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 535 to 553.

[0280] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from an IDH1 protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 535 to 553.

[0281] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 535 to 553.

[0282] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 535 to 553. In some embodiments, one or more of the peptides is a modified or unmodified fragment of a mutated IDH1 protein.

[0283] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 554 to 615.

[0284] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a KRAS protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 554 to 615.

[0285] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 554 to 615.

[0286] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 554 to 615. In some embodiments, one or more of the peptides is a modified or unmodified fragment of a mutated KRAS protein.

[0287] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 616 to 645.

[0288] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from an NRAS protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 616 to 645.

[0289] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 616 to 645.

[0290] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 616 to 645. In some embodiments, one or more of the peptides is a modified or unmodified fragment of a mutated NRAS protein.

[0291] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOS: 646 to 675.

[0292] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a PIK3CA protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 646 to 675.

[0293] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 646 to 675.

[0294] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 646 to 675. In some embodiments, one or more of the peptides is a modified or unmodified fragment of a mutated PIK3CA protein.

[0295] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOS: 676 to 690.

[0296] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a PTEN protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 676 to 690.

[0297] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 676 to 690.

[0298] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 676 to 690. In some embodiments, one or more of the peptides is a modified or unmodified fragment of a mutated PTEN protein.

[0299] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOS: 691 to 758.

[0300] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a TP53 protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 691 to 758.

[0301] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOS: 691 to 758.

[0302] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 691 to 758. In some embodiments, one or more of the peptides is a modified or unmodified fragment of a mutated TP53 protein.

[0303] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 554 to 645.

[0304] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a RAS protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 554 to 645.

[0305] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 554 to 645.

[0306] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 554 to 645. In some embodiments, one or more of the peptides is a modified or unmodified fragment of a mutated RAS protein.

[0307] In another aspect, the present invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 484 to 494.

[0308] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a BRAF V600E protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 484 to 494.

[0309] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 484 to 494.

[0310] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 484 to 494. In some embodiments, the immunogenic peptide composition comprises peptides derived from a BRAF V600E protein mutation.

[0311] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 495 to 502.

[0312] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a BRAF V600M protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 495 to 502.

[0313] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 495 to 502.

[0314] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 495 to 502. In some embodiments, the immunogenic peptide composition comprises peptides derived from a BRAF V600M protein mutation.

[0315] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 503 to 509.

[0316] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from an EGFR A289V protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 503 to 509.

[0317] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 503 to 509.

[0318] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 503 to 509. In some embodiments, the immunogenic peptide composition comprises peptides derived from the EGFR A289V protein mutation.

[0319] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 510 to 519.

[0320] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from an EGFR G598V protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 510 to 519.

[0321] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 510 to 519.

[0322] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 510 to 519. In some embodiments, the immunogenic peptide composition comprises a peptide derived from the EGFR G598V protein mutation.

[0323] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 520 to 527.

[0324] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from an EGFR L858R protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 520 to 527.

[0325] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 520 to 527.

[0326] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 520 to 527. In some embodiments, the immunogenic peptide composition comprises peptides derived from the EGFR L858R protein mutation.

[0327] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 543 to 553.

[0328] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from an IDH1 R132H protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 543 to 553.

[0329] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 543 to 553.

[0330] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 543 to 553. In some embodiments, the immunogenic peptide composition comprises peptides derived from an IDH1 R132H protein mutation.

[0331] In another aspect, the present invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 535 to 542.

[0332] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from an IDH1 R132C protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 535 to 542.

[0333] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 535 to 542.

[0334] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 535 to 542. In some embodiments, the immunogenic peptide composition comprises peptides derived from the IDH1 R132C protein mutation.

[0335] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 569 to 577.

[0336] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a KRAS G12D protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 569 to 577.

[0337] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 569 to 577.

[0338] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 569 to 577. In some embodiments, the immunogenic peptide composition comprises peptides derived from a KRAS G12D protein mutation.

[0339] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOS: 596 to 605.

[0340] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a KRAS G12V protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 596 to 605.

[0341] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 596 to 605.

[0342] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 596 to 605. In some embodiments, the immunogenic peptide composition comprises a peptide derived from a KRAS G12V protein mutation.

[0343] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 578 to 587.

[0344] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a KRAS G12R protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 578 to 587.

[0345] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 578 to 587.

[0346] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 578 to 587. In some embodiments, the immunogenic peptide composition comprises peptides derived from a KRAS G12R protein mutation.

[0347] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 561 to 568.

[0348] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a KRAS G12C protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 561 to 568.

[0349] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 561 to 568.

[0350] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 561 to 568. In some embodiments, the immunogenic peptide composition comprises peptides derived from a KRAS G12C protein mutation.

[0351] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 606 to 615.

[0352] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a KRAS G13D protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 606 to 615.

[0353] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 606 to 615.

[0354] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 606 to 615. In some embodiments, the immunogenic peptide composition comprises a peptide derived from a KRAS G13D protein mutation.

[0355] In another aspect, the present invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 554 to 560.

[0356] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a KRAS G12A protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 554 to 560.

[0357] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 554 to 560.

[0358] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 554 to 560. In some embodiments, the immunogenic peptide composition comprises peptides derived from a KRAS G12A protein mutation.

[0359] In another aspect, the present invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 588 to 595.

[0360] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a KRAS G12S protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 588 to 595.

[0361] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 588 to 595.

[0362] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 588 to 595. In some embodiments, the immunogenic peptide composition comprises peptides derived from a KRAS G12S protein mutation.

[0363] In another aspect, the present invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 634 to 645.

[0364] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from the NRAS Q61R protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 634 to 645.

[0365] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 634 to 645.

[0366] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 634 to 645. In some embodiments, the immunogenic peptide composition comprises peptides derived from the NRAS Q61R protein mutation.

[0367] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 616 to 624.

[0368] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from the NRAS Q61K protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 616 to 624.

[0369] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 616 to 624.

[0370] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 616 to 624. In some embodiments, the immunogenic peptide composition comprises peptides derived from the NRAS Q61K protein mutation.

[0371] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 625 to 633.

[0372] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from an NRAS Q61L protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 625 to 633.

[0373] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 625 to 633.

[0374] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 625 to 633. In some embodiments, the immunogenic peptide composition comprises peptides derived from the NRAS Q61L protein mutation.

[0375] In another aspect, the present invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOS: 646 to 650.

[0376] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a PIK3CA E542K protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 646 to 650.

[0377] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 646 to 650.

[0378] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 646 to 650. In some embodiments, the immunogenic peptide composition comprises peptides derived from the PIK3CA E542K protein mutation.

[0379] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 651 to 657.

[0380] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a PIK3CA E545K protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 651 to 657.

[0381] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 651 to 657.

[0382] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 651 to 657. In some embodiments, the immunogenic peptide composition comprises peptides derived from a PIK3CA E545K protein mutation.

[0383] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 658 to 667.

[0384] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a PIK3CA H1047R protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 658 to 667.

[0385] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 658 to 667.

[0386] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 658 to 667. In some embodiments, the immunogenic peptide composition comprises peptides derived from the PIK3CA H1047R protein mutation.

[0387] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 700 to 707.

[0388] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a TP53 R158L protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 700 to 707.

[0389] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 700 to 707.

[0390] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 700 to 707. In some embodiments, the immunogenic peptide composition comprises a peptide derived from the TP53 R158L protein mutation.

[0391] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 708 to 717.

[0392] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a TP53 R175H protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 708 to 717.

[0393] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 708 to 717.

[0394] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 708 to 717. In some embodiments, the immunogenic peptide composition comprises peptides derived from the TP53 R175H protein mutation.

[0395] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 718 to 723.

[0396] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a TP53 R248Q protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 718 to 723.

[0397] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 718 to 723.

[0398] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 718 to 723. In some embodiments, the immunogenic peptide composition comprises peptides derived from the TP53 R248Q protein mutation.

[0399] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 733 to 739.

[0400] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from the TP53 R273C protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 733 to 739.

[0401] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 733 to 739.

[0402] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 733 to 739. In some embodiments, the immunogenic peptide composition comprises peptides derived from the TP53 R273C protein mutation.

[0403] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 740 to 748.

[0404] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a TP53 R273H protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 740 to 748.

[0405] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 740 to 748.

[0406] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 740 to 748. In some embodiments, the immunogenic peptide composition comprises peptides derived from the TP53 R273H protein mutation.

[0407] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 724 to 732.

[0408] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from the TP53 R248W protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 724 to 732.

[0409] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 724 to 732.

[0410] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 724 to 732. In some embodiments, the immunogenic peptide composition comprises peptides derived from the TP53 R248W protein mutation.

[0411] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 749 to 750.

[0412] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from the TP53 R282W protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 749 to 750.

[0413] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 749 to 750.

[0414] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 749 to 750. In some embodiments, the immunogenic peptide composition comprises peptides derived from the TP53 R282W protein mutation.

[0415] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 751 to 758.

[0416] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a TP53 Y220C protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 751 to 758.

[0417] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 751 to 758.

[0418] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 751 to 758. In some embodiments, the immunogenic peptide composition comprises peptides derived from the TP53 Y220C protein mutation.

[0419] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 668 to 675.

[0420] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a PIK3CA R88Q protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 668 to 675.

[0421] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 668 to 675.

[0422] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 668 to 675. In some embodiments, the immunogenic peptide composition comprises peptides derived from a PIK3CA R88Q protein mutation.

[0423] In another aspect, the present invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 528 to 534.

[0424] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a GTF2I L424H protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 528 to 534.

[0425] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 528 to 534.

[0426] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 528 to 534. In some embodiments, the immunogenic peptide composition comprises peptides derived from the GTF2I L424H protein mutation.

[0427] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 681 to 690.

[0428] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a PTEN R130Q protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 681 to 690.

[0429] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 681 to 690.

[0430] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 681 to 690. In some embodiments, the immunogenic peptide composition comprises peptides derived from the PTEN R130Q protein mutation.

[0431] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 475 to 483.

[0432] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from an AKT1 E17K protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 475 to 483.

[0433] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 475 to 483.

[0434] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 475 to 483. In some embodiments, the immunogenic peptide composition comprises peptides derived from the AKT1 E17K protein mutation.

[0435] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 676 to 680.

[0436] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a PTEN R130G protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 676 to 680.

[0437] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 676 to 680.

[0438] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 676 to 680. In some embodiments, the immunogenic peptide composition comprises peptides derived from the PTEN R130G protein mutation.

[0439] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 691 to 699.

[0440] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from the TP53 H179R protein mutation. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 691 to 699.

[0441] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 691 to 699.

[0442] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 691 to 699. In some embodiments, the immunogenic peptide composition comprises peptides derived from the TP53 H179R protein mutation.

[0443] In another aspect, the present invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 569 to 574, SEQ ID NOs: 578 to 584, SEQ ID NOs: 596 to 599, and SEQ ID NOs: 601 to 603.

[0444] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOS: 569 to 574, 578 to 584, 596 to 599, and 601 to 603. In some embodiments, the nucleic acid sequence is administered as a construct for expression in vivo. In some embodiments, the in vivo administration of the nucleic acid sequence is configured to produce one or more peptides that are presented by HLA class II molecules. In some embodiments, the one or more peptides are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, and TP53. In some embodiments, the one or more peptides are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 The nucleic acid sequence is a modified or unmodified fragment of a protein having a mutation selected from the group consisting of TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the nucleic acid sequence is administered to a subject in an effective amount to prevent cancer. In some embodiments, the nucleic acid sequence is administered to a subject in an effective amount to treat cancer. In some embodiments, the cancer is pancreatic cancer.

[0445] In another aspect, the present invention provides a method of treating or preventing pancreatic cancer by administering to a subject an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOS: 569 to 574, 578 to 584, 596 to 599, and 601 to 603.

[0446] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 569 to 574, SEQ ID NOs: 578 to 584, SEQ ID NOs: 596 to 599, and SEQ ID NOs: 601 to 603.

[0447] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 569 to 574, SEQ ID NOs: 578 to 584, SEQ ID NOs: 596 to 599, and SEQ ID NOs: 601 to 603.

[0448] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOS: 569 to 574, 578 to 584, 596 to 599, and 601 to 603. In some embodiments, the peptides in the immunogenic peptide composition are presented by HLA class II molecules. In some embodiments, the peptides in the immunogenic peptide composition are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, and TP53. In some embodiments, the peptides in the immunogenic peptide composition are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 The modified or unmodified fragment of the protein comprising a mutation selected from the group consisting of TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the immunogenic peptide composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the immunogenic peptide composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the cancer is pancreatic cancer.

[0449] In another aspect, the present invention provides a method of treating or preventing pancreatic cancer by administering to a subject an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOS: 569 to 574, 578 to 584, 596 to 599, and 601 to 603.

[0450] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 569 to 574, SEQ ID NOs: 578 to 584, SEQ ID NOs: 596 to 599, and SEQ ID NOs: 601 to 603.

[0451] In another aspect, the present invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 484 to 485, SEQ ID NOs: 488 to 489, SEQ ID NOs: 495 to 499, SEQ ID NO: 616, SEQ ID NOs: 619 to 620, SEQ ID NOs: 625 to 628, SEQ ID NO: 634, SEQ ID NO: 637, SEQ ID NOs: 639 to 640.

[0452] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 484 to 485, SEQ ID NOs: 488 to 489, SEQ ID NOs: 495 to 499, SEQ ID NOs: 616, SEQ ID NOs: 619 to 620, SEQ ID NOs: 625 to 628, SEQ ID NOs: 634, SEQ ID NO: 637, SEQ ID NOs: 639 to 640. In some embodiments, the nucleic acid sequence is administered as a construct for expression in vivo. In some embodiments, the administration of the nucleic acid sequence in vivo is configured to produce one or more peptides that are presented by HLA class II molecules. In some embodiments, the one or more peptides are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, TP53. In some embodiments, the one or more peptides are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 The nucleic acid sequence is a modified or unmodified fragment of a protein having a mutation selected from the group consisting of TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the nucleic acid sequence is administered to a subject in an effective amount to prevent cancer. In some embodiments, the nucleic acid sequence is administered to a subject in an effective amount to treat cancer. In some embodiments, the cancer is skin cancer.

[0453] In another aspect, the present invention provides a method of treating or preventing skin cancer by administering to a subject an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 484 to 485, SEQ ID NOs: 488 to 489, SEQ ID NOs: 495 to 499, SEQ ID NO: 616, SEQ ID NOs: 619 to 620, SEQ ID NOs: 625 to 628, SEQ ID NO: 634, SEQ ID NO: 637, and SEQ ID NOs: 639 to 640.

[0454] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 484 to 485, SEQ ID NOs: 488 to 489, SEQ ID NOs: 495 to 499, SEQ ID NO: 616, SEQ ID NOs: 619 to 620, SEQ ID NOs: 625 to 628, SEQ ID NO: 634, SEQ ID NO: 637, and SEQ ID NOs: 639 to 640.

[0455] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 484 to 485, SEQ ID NOs: 488 to 489, SEQ ID NOs: 495 to 499, SEQ ID NO: 616, SEQ ID NOs: 619 to 620, SEQ ID NOs: 625 to 628, SEQ ID NO: 634, SEQ ID NO: 637, SEQ ID NOs: 639 to 640.

[0456] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 484 to 485, SEQ ID NOs: 488 to 489, SEQ ID NOs: 495 to 499, SEQ ID NO: 616, SEQ ID NOs: 619 to 620, SEQ ID NOs: 625 to 628, SEQ ID NO: 634, SEQ ID NO: 637, SEQ ID NOs: 639 to 640. In some embodiments, the peptides in the immunogenic peptide composition are presented by HLA class II molecules. In some embodiments, the peptides in the immunogenic peptide composition are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, TP53. In some embodiments, the peptides in the immunogenic peptide composition are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 The modified or unmodified fragment of the protein comprising a mutation selected from the group consisting of TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the immunogenic peptide composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the immunogenic peptide composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the cancer is skin cancer.

[0457] In another aspect, the present invention provides a method of treating or preventing skin cancer by administering to a subject an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 484 to 485, SEQ ID NOs: 488 to 489, SEQ ID NOs: 495 to 499, SEQ ID NO: 616, SEQ ID NOs: 619 to 620, SEQ ID NOs: 625 to 628, SEQ ID NO: 634, SEQ ID NO: 637, and SEQ ID NOs: 639 to 640.

[0458] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 484 to 485, SEQ ID NOs: 488 to 489, SEQ ID NOs: 495 to 499, SEQ ID NO: 616, SEQ ID NOs: 619 to 620, SEQ ID NOs: 625 to 628, SEQ ID NO: 634, SEQ ID NO: 637, and SEQ ID NOs: 639 to 640.

[0459] In another aspect, the present invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 484 to 485, SEQ ID NOs: 488 to 489, SEQ ID NO: 616, SEQ ID NO: 619 to 620, SEQ ID NO: 634, SEQ ID NO: 637, SEQ ID NO: 640.

[0460] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 484 to 485, SEQ ID NOs: 488 to 489, SEQ ID NO: 616, SEQ ID NOs: 619 to 620, SEQ ID NO: 634, SEQ ID NO: 637, SEQ ID NO: 640. In some embodiments, the nucleic acid sequence is administered as a construct for expression in vivo. In some embodiments, the in vivo administration of the nucleic acid sequence is configured to produce one or more peptides that are presented by HLA class II molecules. In some embodiments, the one or more peptides are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, TP53. In some embodiments, the one or more peptides are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 The nucleic acid sequence is a modified or unmodified fragment of a protein having a mutation selected from the group consisting of TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the nucleic acid sequence is administered to a subject in an effective amount to prevent cancer. In some embodiments, the nucleic acid sequence is administered to a subject in an effective amount to treat cancer. In some embodiments, the cancer is thyroid cancer.

[0461] In another aspect, the present invention provides a method of treating or preventing thyroid cancer by administering to a subject an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 484 to 485, SEQ ID NOs: 488 to 489, SEQ ID NO: 616, SEQ ID NO: 619 to 620, SEQ ID NO: 634, SEQ ID NO: 637, and SEQ ID NO: 640.

[0462] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 484 to 485, SEQ ID NOs: 488 to 489, SEQ ID NO: 616, SEQ ID NO: 619 to 620, SEQ ID NO: 634, SEQ ID NO: 637, and SEQ ID NO: 640.

[0463] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 484 to 485, SEQ ID NOs: 488 to 489, SEQ ID NO: 616, SEQ ID NO: 619 to 620, SEQ ID NO: 634, SEQ ID NO: 637, and SEQ ID NO: 640.

[0464] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 484 to 485, SEQ ID NOs: 488 to 489, SEQ ID NO: 616, SEQ ID NOs: 619 to 620, SEQ ID NO: 634, SEQ ID NO: 637, SEQ ID NO: 640. In some embodiments, the peptides in the immunogenic peptide composition are presented by HLA class II molecules. In some embodiments, the peptides in the immunogenic peptide composition are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, TP53. In some embodiments, the peptides in the immunogenic peptide composition are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 The modified or unmodified fragment of the protein comprising a mutation selected from the group consisting of TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the immunogenic peptide composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the immunogenic peptide composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the cancer is thyroid cancer.

[0465] In another aspect, the present invention provides a method of treating or preventing thyroid cancer by administering to a subject an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 484 to 485, SEQ ID NOs: 488 to 489, SEQ ID NO: 616, SEQ ID NO: 619 to 620, SEQ ID NO: 634, SEQ ID NO: 637, and SEQ ID NO: 640.

[0466] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 484 to 485, SEQ ID NOs: 488 to 489, SEQ ID NO: 616, SEQ ID NO: 619 to 620, SEQ ID NO: 634, SEQ ID NO: 637, and SEQ ID NO: 640.

[0467] In another aspect, the present invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NO:508, SEQ ID NO:510, SEQ ID NO:512, SEQ ID NO:514, SEQ ID NOs:535 to 537, SEQ ID NO:539, SEQ ID NOs:543 to 551, SEQ ID NO:708, SEQ ID NO:712, SEQ ID NO:738.

[0468] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NO:508, SEQ ID NO:510, SEQ ID NO:512, SEQ ID NO:514, SEQ ID NO:535 to 537, SEQ ID NO:539, SEQ ID NO:543 to 551, SEQ ID NO:708, SEQ ID NO:712, SEQ ID NO:738. In some embodiments, the nucleic acid sequence is administered as a construct for expression in vivo. In some embodiments, the in vivo administration of the nucleic acid sequence is configured to produce one or more peptides that are presented by HLA class II molecules. In some embodiments, the one or more peptides are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, TP53. In some embodiments, the one or more peptides are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 The nucleic acid sequence is a modified or unmodified fragment of a protein having a mutation selected from the group consisting of TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the nucleic acid sequence is administered to a subject in an effective amount to prevent cancer. In some embodiments, the nucleic acid sequence is administered to a subject in an effective amount to treat cancer. In some embodiments, the cancer is a brain tumor.

[0469] In another aspect, the present invention provides a method of treating or preventing brain tumors by administering to a subject an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NO:508, SEQ ID NO:510, SEQ ID NO:512, SEQ ID NO:514, SEQ ID NOs:535 to 537, SEQ ID NO:539, SEQ ID NOs:543 to 551, SEQ ID NO:708, SEQ ID NO:712, and SEQ ID NO:738.

[0470] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NO:508, SEQ ID NO:510, SEQ ID NO:512, SEQ ID NO:514, SEQ ID NO:535 to 537, SEQ ID NO:539, SEQ ID NO:543 to 551, SEQ ID NO:708, SEQ ID NO:712, and SEQ ID NO:738.

[0471] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NO:508, SEQ ID NO:510, SEQ ID NO:512, SEQ ID NO:514, SEQ ID NOs:535 to 537, SEQ ID NO:539, SEQ ID NOs:543 to 551, SEQ ID NO:708, SEQ ID NO:712, SEQ ID NO:738.

[0472] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NO:508, SEQ ID NO:510, SEQ ID NO:512, SEQ ID NO:514, SEQ ID NO:535 to 537, SEQ ID NO:539, SEQ ID NO:543 to 551, SEQ ID NO:708, SEQ ID NO:712, SEQ ID NO:738. In some embodiments, the peptides in the immunogenic peptide composition are presented by HLA class II molecules. In some embodiments, the peptides in the immunogenic peptide composition are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, TP53. In some embodiments, the peptides in the immunogenic peptide composition are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 The modified or unmodified fragment of the protein comprising a mutation selected from the group consisting of TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the immunogenic peptide composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the immunogenic peptide composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the cancer is a brain tumor.

[0473] In another aspect, the present invention provides a method of treating or preventing brain tumors by administering to a subject an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NO:508, SEQ ID NO:510, SEQ ID NO:512, SEQ ID NO:514, SEQ ID NOs:535 to 537, SEQ ID NO:539, SEQ ID NOs:543 to 551, SEQ ID NO:708, SEQ ID NO:712, and SEQ ID NO:738.

[0474] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NO:508, SEQ ID NO:510, SEQ ID NO:512, SEQ ID NO:514, SEQ ID NO:535 to 537, SEQ ID NO:539, SEQ ID NO:543 to 551, SEQ ID NO:708, SEQ ID NO:712, SEQ ID NO:738.

[0475] In another aspect, the present invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 484 to 485, SEQ ID NOs: 488 to 489, SEQ ID NOs: 569 to 575, SEQ ID NOs: 596 to 599, SEQ ID NOs: 601 to 604, SEQ ID NOs: 606 to 612, SEQ ID NO: 656, SEQ ID NO: 708, SEQ ID NO: 712.

[0476] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 484 to 485, SEQ ID NOs: 488 to 489, SEQ ID NOs: 569 to 575, SEQ ID NOs: 596 to 599, SEQ ID NOs: 601 to 604, SEQ ID NOs: 606 to 612, SEQ ID NO: 656, SEQ ID NO: 708, SEQ ID NO: 712. In some embodiments, the nucleic acid sequence is administered as a construct for expression in vivo. In some embodiments, the administration of the nucleic acid sequence in vivo is configured to produce one or more peptides that are presented by HLA class II molecules. In some embodiments, the one or more peptides are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, TP53. In some embodiments, the one or more peptides are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 The nucleic acid sequence is a modified or unmodified fragment of a protein having a mutation selected from the group consisting of TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the nucleic acid sequence is administered to a subject in an effective amount to prevent cancer. In some embodiments, the nucleic acid sequence is administered to a subject in an effective amount to treat cancer. In some embodiments, the cancer is colorectal cancer.

[0477] In another aspect, the present invention provides a method of treating or preventing colorectal cancer by administering to a subject an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 484 to 485, SEQ ID NOs: 488 to 489, SEQ ID NOs: 569 to 575, SEQ ID NOs: 596 to 599, SEQ ID NOs: 601 to 604, SEQ ID NOs: 606 to 612, SEQ ID NO: 656, SEQ ID NO: 708, SEQ ID NO: 712.

[0478] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 484 to 485, SEQ ID NOs: 488 to 489, SEQ ID NOs: 569 to 575, SEQ ID NOs: 596 to 599, SEQ ID NOs: 601 to 604, SEQ ID NOs: 606 to 612, SEQ ID NO: 656, SEQ ID NO: 708, and SEQ ID NO: 712.

[0479] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 484 to 485, SEQ ID NOs: 488 to 489, SEQ ID NOs: 569 to 575, SEQ ID NOs: 596 to 599, SEQ ID NOs: 601 to 604, SEQ ID NOs: 606 to 612, SEQ ID NO: 656, SEQ ID NO: 708, SEQ ID NO: 712.

[0480] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 484 to 485, SEQ ID NOs: 488 to 489, SEQ ID NOs: 569 to 575, SEQ ID NOs: 596 to 599, SEQ ID NOs: 601 to 604, SEQ ID NOs: 606 to 612, SEQ ID NO: 656, SEQ ID NO: 708, SEQ ID NO: 712. In some embodiments, the peptides in the immunogenic peptide composition are presented by HLA class II molecules. In some embodiments, the peptides in the immunogenic peptide composition are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, TP53. In some embodiments, the peptides in the immunogenic peptide composition are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 The modified or unmodified fragment of the protein comprising a mutation selected from the group consisting of TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the immunogenic peptide composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the immunogenic peptide composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the cancer is colorectal cancer.

[0481] In another aspect, the present invention provides a method of treating or preventing colorectal cancer by administering to a subject an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 484 to 485, SEQ ID NOs: 488 to 489, SEQ ID NOs: 569 to 575, SEQ ID NOs: 596 to 599, SEQ ID NOs: 601 to 604, SEQ ID NOs: 606 to 612, SEQ ID NO: 656, SEQ ID NO: 708, SEQ ID NO: 712.

[0482] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 484 to 485, SEQ ID NOs: 488 to 489, SEQ ID NOs: 569 to 575, SEQ ID NOs: 596 to 599, SEQ ID NOs: 601 to 604, SEQ ID NOs: 606 to 612, SEQ ID NO: 656, SEQ ID NO: 708, and SEQ ID NO: 712.

[0483] In another aspect, the present invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs:520 to 521, SEQ ID NOs:523 to 524, SEQ ID NOs:554 to 556, SEQ ID NO:558, SEQ ID NO:561, SEQ ID NOs:563 to 565, SEQ ID NOs:569 to 573, SEQ ID NOs:596 to 600, SEQ ID NO:650, SEQ ID NO:656, SEQ ID NOs:700 to 705.

[0484] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 520 to 521, SEQ ID NOs: 523 to 524, SEQ ID NOs: 554 to 556, SEQ ID NO: 558, SEQ ID NO: 561, SEQ ID NOs: 563 to 565, SEQ ID NOs: 569 to 573, SEQ ID NOs: 596 to 600, SEQ ID NO: 650, SEQ ID NO: 656, SEQ ID NOs: 700 to 705. In some embodiments, the nucleic acid sequence is administered as a construct for expression in vivo. In some embodiments, the in vivo administration of the nucleic acid sequence is configured to produce one or more peptides that are presented by an HLA class II molecule. In some embodiments, the one or more peptides are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, TP53. In some embodiments, the one or more peptides are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C, or a modified or unmodified fragment of a protein having a mutation selected from the group consisting of: TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C, In some embodiments, the nucleic acid sequence is administered to a subject in an effective amount to prevent cancer.In some embodiments, the nucleic acid sequence is administered to a subject in an effective amount to treat cancer, hi some embodiments, the cancer is bronchial and lung cancer.

[0485] In another aspect, the present invention provides a method of treating or preventing bronchial and lung cancer by administering to a subject an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs:520 to 521, SEQ ID NOs:523 to 524, SEQ ID NOs:554 to 556, SEQ ID NO:558, SEQ ID NO:561, SEQ ID NOs:563 to 565, SEQ ID NOs:569 to 573, SEQ ID NOs:596 to 600, SEQ ID NO:650, SEQ ID NO:656, and SEQ ID NOs:700 to 705.

[0486] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 520 to 521, SEQ ID NOs: 523 to 524, SEQ ID NOs: 554 to 556, SEQ ID NO: 558, SEQ ID NO: 561, SEQ ID NOs: 563 to 565, SEQ ID NOs: 569 to 573, SEQ ID NOs: 596 to 600, SEQ ID NO: 650, SEQ ID NO: 656, and SEQ ID NOs: 700 to 705.

[0487] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs:520 to 521, SEQ ID NOs:523 to 524, SEQ ID NOs:554 to 556, SEQ ID NO:558, SEQ ID NO:561, SEQ ID NOs:563 to 565, SEQ ID NOs:569 to 573, SEQ ID NOs:596 to 600, SEQ ID NO:650, SEQ ID NO:656, SEQ ID NOs:700 to 705.

[0488] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs:520 to 521, SEQ ID NOs:523 to 524, SEQ ID NOs:554 to 556, SEQ ID NO:558, SEQ ID NO:561, SEQ ID NOs:563 to 565, SEQ ID NOs:569 to 573, SEQ ID NOs:596 to 600, SEQ ID NO:650, SEQ ID NO:656, SEQ ID NOs:700 to 705. In some embodiments, the peptides in the immunogenic peptide composition are presented by HLA class II molecules. In some embodiments, the peptides in the immunogenic peptide composition are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, TP53. In some embodiments, the peptides in the immunogenic peptide composition are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 The modified or unmodified fragment of the protein comprising a mutation selected from the group consisting of TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the immunogenic peptide composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the immunogenic peptide composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the cancer is bronchial cancer and lung cancer.

[0489] In another aspect, the present invention provides a method of treating or preventing bronchial and lung cancer by administering to a subject an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs:520 to 521, SEQ ID NOs:523 to 524, SEQ ID NOs:554 to 556, SEQ ID NO:558, SEQ ID NO:561, SEQ ID NOs:563 to 565, SEQ ID NOs:569 to 573, SEQ ID NOs:596 to 600, SEQ ID NO:650, SEQ ID NO:656, and SEQ ID NOs:700 to 705.

[0490] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 520 to 521, SEQ ID NOs: 523 to 524, SEQ ID NOs: 554 to 556, SEQ ID NO: 558, SEQ ID NO: 561, SEQ ID NOs: 563 to 565, SEQ ID NOs: 569 to 573, SEQ ID NOs: 596 to 600, SEQ ID NO: 650, SEQ ID NO: 656, and SEQ ID NOs: 700 to 705.

[0491] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 646 to 667, SEQ ID NOs: 708 to 717, and SEQ ID NOs: 733 to 748.

[0492] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOS: 646 to 667, SEQ ID NOS: 708 to 717, and SEQ ID NOS: 733 to 748. In some embodiments, the nucleic acid sequence is administered as a construct for expression in vivo. In some embodiments, the in vivo administration of the nucleic acid sequence is configured to produce one or more peptides that are presented by HLA class II molecules. In some embodiments, the one or more peptides are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, and TP53. In some embodiments, the one or more peptides are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 The nucleic acid sequence is a modified or unmodified fragment of a protein having a mutation selected from the group consisting of TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the nucleic acid sequence is administered to a subject in an effective amount to prevent cancer. In some embodiments, the nucleic acid sequence is administered to a subject in an effective amount to treat cancer. In some embodiments, the cancer is breast cancer.

[0493] In another aspect, the invention provides a method of treating or preventing breast cancer by administering to a subject an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOS: 646 to 667, 708 to 717, and 733 to 748.

[0494] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 646 to 667, SEQ ID NOs: 708 to 717, and SEQ ID NOs: 733 to 748.

[0495] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOS: 646 to 667, 708 to 717, and 733 to 748.

[0496] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOS: 646 to 667, SEQ ID NOS: 708 to 717, and SEQ ID NOS: 733 to 748. In some embodiments, the peptides in the immunogenic peptide composition are presented by HLA class II molecules. In some embodiments, the peptides in the immunogenic peptide composition are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, and TP53. In some embodiments, the peptides in the immunogenic peptide composition are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 The modified or unmodified fragment of the protein comprising a mutation selected from the group consisting of TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the immunogenic peptide composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the immunogenic peptide composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the cancer is breast cancer.

[0497] In another aspect, the invention provides a method of treating or preventing breast cancer by administering to a subject an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOS: 646 to 667, 708 to 717, and 733 to 748.

[0498] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 646 to 667, SEQ ID NOs: 708 to 717, and SEQ ID NOs: 733 to 748.

[0499] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 646 to 667, SEQ ID NOs: 708 to 717, and SEQ ID NOs: 733 to 748.

[0500] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOS: 646 to 667, SEQ ID NOS: 708 to 717, and SEQ ID NOS: 733 to 748. In some embodiments, the nucleic acid sequence is administered as a construct for expression in vivo. In some embodiments, the in vivo administration of the nucleic acid sequence is configured to produce one or more peptides that are presented by HLA class II molecules. In some embodiments, the one or more peptides are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, and TP53. In some embodiments, the one or more peptides are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 The nucleic acid sequence is a modified or unmodified fragment of a protein having a mutation selected from the group consisting of TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the nucleic acid sequence is administered to a subject in an effective amount to prevent cancer. In some embodiments, the nucleic acid sequence is administered to a subject in an effective amount to treat cancer. In some embodiments, the cancer is ovarian cancer.

[0501] In another aspect, the present invention provides a method of treating or preventing ovarian cancer by administering to a subject an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOS: 646 to 667, SEQ ID NOS: 708 to 717, and SEQ ID NOS: 733 to 748.

[0502] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 646 to 667, SEQ ID NOs: 708 to 717, and SEQ ID NOs: 733 to 748.

[0503] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOS: 646 to 667, 708 to 717, and 733 to 748.

[0504] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOS: 646 to 667, SEQ ID NOS: 708 to 717, and SEQ ID NOS: 733 to 748. In some embodiments, the peptides in the immunogenic peptide composition are presented by HLA class II molecules. In some embodiments, the peptides in the immunogenic peptide composition are modified or unmodified fragments of mutant proteins selected from the group consisting of AKT1, BRAF, EGFR, GTF2I, HRAS, IDH1, KRAS, NRAS, PIK3CA, PTEN, and TP53. In some embodiments, the peptides in the immunogenic peptide composition are selected from the group consisting of AKT1 E17K, BRAF V600E, BRAF V600M, EGFR A289V, EGFR G598V, EGFR L858R, GTF2I L424H, IDH1 R132C, IDH1 R132H, KRAS G12A, KRAS G12C, KRAS G12D, KRAS G12R, KRAS G12S, KRAS G12V, KRAS G13D, NRAS Q61K, NRAS Q61L, NRAS Q61R, PIK3CA E542K, PIK3CA E545K, PIK3CA H1047R, PIK3CA R88Q, PTEN R130G, PTEN R130Q, TP53 H179R, TP53 R158L, TP53 The immunogenic peptide composition is a modified or unmodified fragment of a protein comprising a mutation selected from the group consisting of TP53 R175H, TP53 R248Q, TP53 R248W, TP53 R273C, TP53 R273H, TP53 R282W, TP53 Y220C. In some embodiments, the immunogenic peptide composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the immunogenic peptide composition is administered to a subject in an effective amount to treat cancer. In some embodiments, the cancer is ovarian cancer.

[0505] In another aspect, the present invention provides a method of treating or preventing ovarian cancer by administering to a subject an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOS: 646 to 667, SEQ ID NOS: 708 to 717, and SEQ ID NOS: 733 to 748.

[0506] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 646 to 667, SEQ ID NOs: 708 to 717, and SEQ ID NOs: 733 to 748. CT antigen vaccine

[0507] In one aspect, the invention provides a method for the preparation of a polypeptide comprising SEQ ID NO: 28796 to 28864, SEQ ID NO: 37110 to 37174, SEQ ID NO: 41321 to 41397, SEQ ID NO: 41770, SEQ ID NO: 49004, SEQ ID NO: 49071, SEQ ID NO: 49395, SEQ ID NO: 50632, SEQ ID NO: 50729, SEQ ID NO: 51434 to 51510, SEQ ID NO: 55758, SEQ ID NO: 60456 to 60527, SEQ ID NO: 68238 to 68321, SEQ ID NO: 77091, SEQ ID NO: 77210, SEQ ID NO: 84188, SEQ ID NO: 87951, SEQ ID NO: In one embodiment, nucleic acid sequences are provided that encode one or more amino acid sequences selected from the group consisting of SEQ ID NO:88144, SEQ ID NO:95593 to 95664, SEQ ID NO:113808 to 113869, SEQ ID NO:125134 to 125218, SEQ ID NO:144109 to 144188, SEQ ID NO:162383 to 162453, SEQ ID NO:166444 to 166531, SEQ ID NO:167118, SEQ ID NO:169740, SEQ ID NO:173412, SEQ ID NO:179404, SEQ ID NO:182574 to 182654.

[0508] In some embodiments, the nucleic acid sequence is selected from the group consisting of SEQ ID NOs: 28796 to 28864, SEQ ID NOs: 37110 to 37174, SEQ ID NOs: 41321 to 41397, SEQ ID NO: 41770, SEQ ID NO: 49004, SEQ ID NO: 49071, SEQ ID NO: 49395, SEQ ID NO: 50632, SEQ ID NO: 50729, SEQ ID NOs: 51434 to 51510, SEQ ID NO: 55758, SEQ ID NOs: 60456 to 60527, SEQ ID NOs: 68238 to 68321, SEQ ID NO: 77091, SEQ ID NO: 77210, SEQ ID NO: 84188, SEQ ID NO: 87951, SEQ ID NO: and SEQ ID NO:88144, SEQ ID NO:95593 to 95664, SEQ ID NO:113808 to 113869, SEQ ID NO:125134 to 125218, SEQ ID NO:144109 to 144188, SEQ ID NO:162383 to 162453, SEQ ID NO:166444 to 166531, SEQ ID NO:167118, SEQ ID NO:169740, SEQ ID NO:173412, SEQ ID NO:179404, and SEQ ID NO:182574 to 182654.

[0509] In another aspect, the invention relates to a method for the preparation of a medicament for the treatment of a disease comprising administering to a patient a medicament therapies comprising the steps of: SEQ ID NOs: 28796 to 28864, SEQ ID NOs: 37110 to 37174, SEQ ID NOs: 41321 to 41397, SEQ ID NO: 41770, SEQ ID NO: 49004, SEQ ID NO: 49071, SEQ ID NO: 49395, SEQ ID NO: 50632, SEQ ID NO: 50729, SEQ ID NOs: 51434 to 51510, SEQ ID NO: 55758, SEQ ID NOs: 60456 to 60527, SEQ ID NOs: 68238 to 68321, SEQ ID NO: 77091, SEQ ID NO: 77210, SEQ ID NO: 84188, SEQ ID NO: 87951, SEQ ID NO: In one embodiment, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NO:88144, SEQ ID NO:95593 to 95664, SEQ ID NO:113808 to 113869, SEQ ID NO:125134 to 125218, SEQ ID NO:144109 to 144188, SEQ ID NO:162383 to 162453, SEQ ID NO:166444 to 166531, SEQ ID NO:167118, SEQ ID NO:169740, SEQ ID NO:173412, SEQ ID NO:179404, SEQ ID NO:182574 to 182654.

[0510] In some embodiments, the immunogenic composition is administered to a subject. In some embodiments, the immunogenic composition comprises a polypeptide selected from the group consisting of SEQ ID NOs: 28796 to 28864, SEQ ID NOs: 37110 to 37174, SEQ ID NOs: 41321 to 41397, SEQ ID NO: 41770, SEQ ID NO: 49004, SEQ ID NO: 49071, SEQ ID NO: 49395, SEQ ID NO: 50632, SEQ ID NO: 50729, SEQ ID NOs: 51434 to 51510, SEQ ID NO: 55758, SEQ ID NOs: 60456 to 60527, SEQ ID NOs: 68238 to 68321, SEQ ID NOs: 77091, SEQ ID NO: 77210, SEQ ID NO: 84188, SEQ ID NOs: In some embodiments, the nucleic acid sequence comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NO:87951, SEQ ID NO:88144, SEQ ID NO:95593 to 95664, SEQ ID NO:113808 to 113869, SEQ ID NO:125134 to 125218, SEQ ID NO:144109 to 144188, SEQ ID NO:162383 to 162453, SEQ ID NO:166444 to 166531, SEQ ID NO:167118, SEQ ID NO:169740, SEQ ID NO:173412, SEQ ID NO:179404, SEQ ID NO:182574 to 182654. In some embodiments, the nucleic acid sequence is administered as a construct for expression in vivo. In some embodiments, the in vivo administration of the nucleic acid sequence is configured to produce one or more peptides displayed by HLA class I molecules. In some embodiments, the one or more peptides are modified or unmodified fragments of a protein selected from the group consisting of CTG1B, KKLC1, MAGA1, MAGA3, MAGA4, MAGC1, MAGC3, MAR1, PMEL, PRAME, SSX2, TYRP1, TYRP2. In some embodiments, the immunogenic composition is administered to a subject in an effective amount to prevent cancer. In some embodiments, the immunogenic composition is administered to a subject in an effective amount to treat cancer.In some embodiments, the cancer is selected from the group consisting of pancreatic cancer, skin cancer, thyroid cancer, brain cancer, colorectal cancer, bronchial and lung cancer, breast cancer, and ovarian cancer. In some embodiments, the immunogenic composition comprises a sequence selected from the group consisting of SEQ ID NOs: 28796 to 28864, SEQ ID NOs: 37110 to 37174, SEQ ID NOs: 41321 to 41397, SEQ ID NO: 41770, SEQ ID NO: 49004, SEQ ID NO: 49071, SEQ ID NO: 49395, SEQ ID NO: 50632, SEQ ID NO: 50729, SEQ ID NOs: 51434 to 51510, SEQ ID NO: 55758, SEQ ID NOs: 60456 to 60527, SEQ ID NOs: 68238 to 68321, SEQ ID NOs: 77091, SEQ ID NO: 77210, SEQ ID NO: 84188, SEQ ID NOs: and SEQ ID NO: 87951, SEQ ID NO: 88144, SEQ ID NO: 95593 to 95664, SEQ ID NO: 113808 to 113869, SEQ ID NO: 125134 to 125218, SEQ ID NO: 144109 to 144188, SEQ ID NO: 162383 to 162453, SEQ ID NO: 166444 to 166531, SEQ ID NO: 167118, SEQ ID NO: 169740, SEQ ID NO: 173412, SEQ ID NO: 179404, and SEQ ID NO: 182574 to 182654.

[0511] In another aspect, the invention provides a method for the preparation of a polypeptide comprising SEQ ID NO: 28796 to 28864, SEQ ID NO: 37110 to 37174, SEQ ID NO: 41321 to 41397, SEQ ID NO: 41770, SEQ ID NO: 49004, SEQ ID NO: 49071, SEQ ID NO: 49395, SEQ ID NO: 50632, SEQ ID NO: 50729, SEQ ID NO: 51434 to 51510, SEQ ID NO: 55758, SEQ ID NO: 60456 to 60527, SEQ ID NO: 68238 to 68321, SEQ ID NO: 77091, SEQ ID NO: 77210, SEQ ID NO: 84188, SEQ ID NO: 87951, SEQ ID NO: and SEQ ID NO:88144, SEQ ID NO:95593 to 95664, SEQ ID NO:113808 to 113869, SEQ ID NO:125134 to 125218, SEQ ID NO:144109 to 144188, SEQ ID NO:162383 to 162453, SEQ ID NO:166444 to 166531, SEQ ID NO:167118, SEQ ID NO:169740, SEQ ID NO:173412, SEQ ID NO:179404, SEQ ID NO:182574 to 182654.

[0512] In some embodiments, the immunogenic peptide composition comprises a peptide selected from the group consisting of SEQ ID NOs: 28796 to 28864, SEQ ID NOs: 37110 to 37174, SEQ ID NOs: 41321 to 41397, SEQ ID NO: 41770, SEQ ID NO: 49004, SEQ ID NO: 49071, SEQ ID NO: 49395, SEQ ID NO: 50632, SEQ ID NO: 50729, SEQ ID NOs: 51434 to 51510, SEQ ID NO: 55758, SEQ ID NOs: 60456 to 60527, SEQ ID NOs: 68238 to 68321, SEQ ID NOs: 77091, SEQ ID NO: 77210, SEQ ID NO: 84188, SEQ ID NOs: and SEQ ID NO: 87951, SEQ ID NO: 88144, SEQ ID NO: 95593 to 95664, SEQ ID NO: 113808 to 113869, SEQ ID NO: 125134 to 125218, SEQ ID NO: 144109 to 144188, SEQ ID NO: 162383 to 162453, SEQ ID NO: 166444 to 166531, SEQ ID NO: 167118, SEQ ID NO: 169740, SEQ ID NO: 173412, SEQ ID NO: 179404, and SEQ ID NO: 182574 to 182654.

[0513] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOS: 28796 to 28830.

[0514] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a CTG1B protein, hi some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 28796 to 28830.

[0515] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 28796 to 28830.

[0516] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 28796 to 28830. In some embodiments, one or more of the peptides is a modified or unmodified fragment of the CTG1B protein.

[0517] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 41321 to 41354.

[0518] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a MAGA1 protein. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 41321 to 41354.

[0519] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 41321 to 41354.

[0520] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 41321 to 41354. In some embodiments, one or more of the peptides is a modified or unmodified fragment of the MAGA1 protein.

[0521] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NO: 41770, SEQ ID NO: 49004, SEQ ID NO: 51434 to 51468.

[0522] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a MAGA3 protein, hi some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NO: 41770, SEQ ID NO: 49004, SEQ ID NO: 51434 to 51468.

[0523] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NO: 41770, SEQ ID NO: 49004, SEQ ID NO: 51434 to 51468.

[0524] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NO: 41770, SEQ ID NO: 49004, SEQ ID NO: 51434 to 51468. In some embodiments, one or more of the peptides is a modified or unmodified fragment of the MAGA3 protein.

[0525] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NO: 41352, SEQ ID NO: 41770, and SEQ ID NOs: 60456 to 60487.

[0526] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a MAGA4 protein, hi some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NO: 41352, SEQ ID NO: 41770, and SEQ ID NO: 60456 to 60487.

[0527] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NO: 41352, SEQ ID NO: 41770, and SEQ ID NOs: 60456 to 60487.

[0528] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NO: 41352, SEQ ID NO: 41770, SEQ ID NO: 60456 to 60487. In some embodiments, one or more of the peptides is a modified or unmodified fragment of the MAGA4 protein.

[0529] In another aspect, the present invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NO: 49395 and SEQ ID NO: 68238 to 68272.

[0530] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from the MAGC1 protein, hi some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NO: 49395 and SEQ ID NO: 68238 to 68272.

[0531] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NO: 49395 and SEQ ID NO: 68238 to 68272.

[0532] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NO: 49395 and SEQ ID NO: 68238 to 68272. In some embodiments, one or more of the peptides is a modified or unmodified fragment of the MAGC1 protein.

[0533] In another aspect, the present invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NO: 77091, SEQ ID NO: 77210, SEQ ID NO: 84188, SEQ ID NO: 88144, SEQ ID NO: 95593 to 95624.

[0534] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a MAGC3 protein, hi some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NO: 77091, SEQ ID NO: 77210, SEQ ID NO: 84188, SEQ ID NO: 88144, SEQ ID NO: 95593 to 95624.

[0535] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NO: 77091, SEQ ID NO: 77210, SEQ ID NO: 84188, SEQ ID NO: 88144, SEQ ID NO: 95593 to 95624.

[0536] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NO: 77091, SEQ ID NO: 77210, SEQ ID NO: 84188, SEQ ID NO: 88144, SEQ ID NO: 95593 to 95624. In some embodiments, one or more of the peptides is a modified or unmodified fragment of the MAGC3 protein.

[0537] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOS: 162383 to 162420.

[0538] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from the SSX2 protein. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 162383 to 162420.

[0539] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 162383 to 162420.

[0540] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 162383 to 162420. In some embodiments, one or more of the peptides is a modified or unmodified fragment of the SSX2 protein.

[0541] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 144109 to 144142.

[0542] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a PRAME protein. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 144109 to 144142.

[0543] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 144109 to 144142.

[0544] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 144109 to 144142. In some embodiments, one or more of the peptides is a modified or unmodified fragment of the PRAME protein.

[0545] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOS: 37110 to 37145.

[0546] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from the KKLC1 protein. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 37110 to 37145.

[0547] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 37110 to 37145.

[0548] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 37110 to 37145. In some embodiments, one or more of the peptides is a modified or unmodified fragment of the KKLC1 protein.

[0549] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 125134 to 125167.

[0550] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a PMEL protein, hi some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 125134 to 125167.

[0551] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 125134 to 125167.

[0552] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 125134 to 125167. In some embodiments, one or more of the peptides is a modified or unmodified fragment of the PMEL protein.

[0553] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOS: 166444 to 166480.

[0554] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a TYRP1 protein. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 166444 to 166480.

[0555] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 166444 to 166480.

[0556] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 166444 to 166480. In some embodiments, one or more of the peptides is a modified or unmodified fragment of the TYRP1 protein.

[0557] In another aspect, the present invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NO: 169740, SEQ ID NO: 173412, SEQ ID NO: 179404, SEQ ID NO: 182574 to 182606.

[0558] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a TYRP2 protein, hi some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NO: 169740, SEQ ID NO: 173412, SEQ ID NO: 179404, SEQ ID NO: 182574 to 182606.

[0559] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NO: 169740, SEQ ID NO: 173412, SEQ ID NO: 179404, SEQ ID NO: 182574 to 182606.

[0560] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NO: 169740, SEQ ID NO: 173412, SEQ ID NO: 179404, SEQ ID NO: 182574 to 182606. In some embodiments, one or more of the peptides is a modified or unmodified fragment of the TYRP2 protein.

[0561] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 113808 to 113843.

[0562] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from the MAR1 protein. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 113808 to 113843.

[0563] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 113808 to 113843.

[0564] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 113808 to 113843. In some embodiments, one or more of the peptides is a modified or unmodified fragment of the MAR1 protein.

[0565] In one aspect, the invention provides a method for the preparation of a polypeptide comprising SEQ ID NO: 28796 to 28864, SEQ ID NO: 37110 to 37174, SEQ ID NO: 41321 to 41397, SEQ ID NO: 41770, SEQ ID NO: 49004, SEQ ID NO: 49071, SEQ ID NO: 49395, SEQ ID NO: 50632, SEQ ID NO: 50729, SEQ ID NO: 51434 to 51510, SEQ ID NO: 55758, SEQ ID NO: 60456 to 60527, SEQ ID NO: 68238 to 68321, SEQ ID NO: 77091, SEQ ID NO: 77210, SEQ ID NO: 84188, SEQ ID NO: 87951, SEQ ID NO: In one embodiment, nucleic acid sequences are provided that encode one or more amino acid sequences selected from the group consisting of SEQ ID NO:88144, SEQ ID NO:95593 to 95664, SEQ ID NO:113808 to 113869, SEQ ID NO:125134 to 125218, SEQ ID NO:144109 to 144188, SEQ ID NO:162383 to 162453, SEQ ID NO:166444 to 166531, SEQ ID NO:167118, SEQ ID NO:169740, SEQ ID NO:173412, SEQ ID NO:179404, SEQ ID NO:182574 to 182654.

[0566] In some embodiments, the nucleic acid sequence is selected from the group consisting of SEQ ID NOs: 28796 to 28864, SEQ ID NOs: 37110 to 37174, SEQ ID NOs: 41321 to 41397, SEQ ID NO: 41770, SEQ ID NO: 49004, SEQ ID NO: 49071, SEQ ID NO: 49395, SEQ ID NO: 50632, SEQ ID NO: 50729, SEQ ID NOs: 51434 to 51510, SEQ ID NO: 55758, SEQ ID NOs: 60456 to 60527, SEQ ID NOs: 68238 to 68321, SEQ ID NO: 77091, SEQ ID NO: 77210, SEQ ID NO: 84188, SEQ ID NO: 87951, SEQ ID NO: and SEQ ID NO:88144, SEQ ID NO:95593 to 95664, SEQ ID NO:113808 to 113869, SEQ ID NO:125134 to 125218, SEQ ID NO:144109 to 144188, SEQ ID NO:162383 to 162453, SEQ ID NO:166444 to 166531, SEQ ID NO:167118, SEQ ID NO:169740, SEQ ID NO:173412, SEQ ID NO:179404, and SEQ ID NO:182574 to 182654.

[0567] In another aspect, the invention relates to a method for the preparation of a medicament for the treatment of a disease comprising administering to a patient a medicament therapies comprising the steps of: SEQ ID NOs: 28796 to 28864, SEQ ID NOs: 37110 to 37174, SEQ ID NOs: 41321 to 41397, SEQ ID NO: 41770, SEQ ID NO: 49004, SEQ ID NO: 49071, SEQ ID NO: 49395, SEQ ID NO: 50632, SEQ ID NO: 50729, SEQ ID NOs: 51434 to 51510, SEQ ID NO: 55758, SEQ ID NOs: 60456 to 60527, SEQ ID NOs: 68238 to 68321, SEQ ID NO: 77091, SEQ ID NO: 77210, SEQ ID NO: 84188, SEQ ID NO: 87951, SEQ ID NO: In one embodiment, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NO:88144, SEQ ID NO:95593 to 95664, SEQ ID NO:113808 to 113869, SEQ ID NO:125134 to 125218, SEQ ID NO:144109 to 144188, SEQ ID NO:162383 to 162453, SEQ ID NO:166444 to 166531, SEQ ID NO:167118, SEQ ID NO:169740, SEQ ID NO:173412, SEQ ID NO:179404, SEQ ID NO:182574 to 182654.

[0568] In some embodiments, the immunogenic composition is administered to a subject. In some embodiments, the immunogenic composition comprises a polypeptide selected from the group consisting of SEQ ID NOs: 28796 to 28864, SEQ ID NOs: 37110 to 37174, SEQ ID NOs: 41321 to 41397, SEQ ID NO: 41770, SEQ ID NO: 49004, SEQ ID NO: 49071, SEQ ID NO: 49395, SEQ ID NO: 50632, SEQ ID NO: 50729, SEQ ID NOs: 51434 to 51510, SEQ ID NO: 55758, SEQ ID NOs: 60456 to 60527, SEQ ID NOs: 68238 to 68321, SEQ ID NOs: 77091, SEQ ID NO: 77210, SEQ ID NO: 84188, SEQ ID NOs: In some embodiments, the nucleic acid sequence comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NO:87951, SEQ ID NO:88144, SEQ ID NO:95593 to 95664, SEQ ID NO:113808 to 113869, SEQ ID NO:125134 to 125218, SEQ ID NO:144109 to 144188, SEQ ID NO:162383 to 162453, SEQ ID NO:166444 to 166531, SEQ ID NO:167118, SEQ ID NO:169740, SEQ ID NO:173412, SEQ ID NO:179404, SEQ ID NO:182574 to 182654. In some embodiments, the nucleic acid sequence is administered as a construct for expression in vivo. In some embodiments, the in vivo administration of the nucleic acid sequence is configured to produce one or more peptides that are displayed by an HLA class II molecule. In some embodiments, the one or more peptides are modified or unmodified fragments of a protein selected from the group consisting of CTG1B, KKLC1, MAGA1, MAGA3, MAGA4, MAGC1, MAGC3, MAR1, PMEL, PRAME, SSX2, TYRP1, TYRP2. In some embodiments, the immunogenic composition is administered to a subject in an effective amount to prevent cancer. In some embodiments, the immunogenic composition is administered to a subject in an effective amount to treat cancer.In some embodiments, the cancer is selected from the group consisting of pancreatic cancer, skin cancer, thyroid cancer, brain cancer, colorectal cancer, bronchial and lung cancer, breast cancer, and ovarian cancer. In some embodiments, the immunogenic composition comprises a sequence selected from the group consisting of SEQ ID NOs: 28796 to 28864, SEQ ID NOs: 37110 to 37174, SEQ ID NOs: 41321 to 41397, SEQ ID NO: 41770, SEQ ID NO: 49004, SEQ ID NO: 49071, SEQ ID NO: 49395, SEQ ID NO: 50632, SEQ ID NO: 50729, SEQ ID NOs: 51434 to 51510, SEQ ID NO: 55758, SEQ ID NOs: 60456 to 60527, SEQ ID NOs: 68238 to 68321, SEQ ID NOs: 77091, SEQ ID NO: 77210, SEQ ID NO: 84188, SEQ ID NOs: and SEQ ID NO: 87951, SEQ ID NO: 88144, SEQ ID NO: 95593 to 95664, SEQ ID NO: 113808 to 113869, SEQ ID NO: 125134 to 125218, SEQ ID NO: 144109 to 144188, SEQ ID NO: 162383 to 162453, SEQ ID NO: 166444 to 166531, SEQ ID NO: 167118, SEQ ID NO: 169740, SEQ ID NO: 173412, SEQ ID NO: 179404, and SEQ ID NO: 182574 to 182654.

[0569] In another aspect, the invention provides a method for the preparation of a polypeptide comprising SEQ ID NO: 28796 to 28864, SEQ ID NO: 37110 to 37174, SEQ ID NO: 41321 to 41397, SEQ ID NO: 41770, SEQ ID NO: 49004, SEQ ID NO: 49071, SEQ ID NO: 49395, SEQ ID NO: 50632, SEQ ID NO: 50729, SEQ ID NO: 51434 to 51510, SEQ ID NO: 55758, SEQ ID NO: 60456 to 60527, SEQ ID NO: 68238 to 68321, SEQ ID NO: 77091, SEQ ID NO: 77210, SEQ ID NO: 84188, SEQ ID NO: 87951, SEQ ID NO: and SEQ ID NO:88144, SEQ ID NO:95593 to 95664, SEQ ID NO:113808 to 113869, SEQ ID NO:125134 to 125218, SEQ ID NO:144109 to 144188, SEQ ID NO:162383 to 162453, SEQ ID NO:166444 to 166531, SEQ ID NO:167118, SEQ ID NO:169740, SEQ ID NO:173412, SEQ ID NO:179404, SEQ ID NO:182574 to 182654.

[0570] In some embodiments, the immunogenic peptide composition comprises a peptide selected from the group consisting of SEQ ID NOs: 28796 to 28864, SEQ ID NOs: 37110 to 37174, SEQ ID NOs: 41321 to 41397, SEQ ID NO: 41770, SEQ ID NO: 49004, SEQ ID NO: 49071, SEQ ID NO: 49395, SEQ ID NO: 50632, SEQ ID NO: 50729, SEQ ID NOs: 51434 to 51510, SEQ ID NO: 55758, SEQ ID NOs: 60456 to 60527, SEQ ID NOs: 68238 to 68321, SEQ ID NOs: 77091, SEQ ID NO: 77210, SEQ ID NO: 84188, SEQ ID NOs: and SEQ ID NO: 87951, SEQ ID NO: 88144, SEQ ID NO: 95593 to 95664, SEQ ID NO: 113808 to 113869, SEQ ID NO: 125134 to 125218, SEQ ID NO: 144109 to 144188, SEQ ID NO: 162383 to 162453, SEQ ID NO: 166444 to 166531, SEQ ID NO: 167118, SEQ ID NO: 169740, SEQ ID NO: 173412, SEQ ID NO: 179404, and SEQ ID NO: 182574 to 182654.

[0571] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 197897 to 197901.

[0572] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a CTG1B protein. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 197897 to 197901.

[0573] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 197897 to 197901.

[0574] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 197897 to 197901. In some embodiments, one or more of the peptides is a modified or unmodified fragment of the CTG1B protein.

[0575] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 211901 to 211904.

[0576] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a MAGA1 protein. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 211901 to 211904.

[0577] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 211901 to 211904.

[0578] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 211901 to 211904. In some embodiments, one or more of the peptides is a modified or unmodified fragment of the MAGA1 protein.

[0579] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 223623 to 223627.

[0580] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a MAGA3 protein, hi some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 223623 to 223627.

[0581] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 223623 to 223627.

[0582] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 223623 to 223627. In some embodiments, one or more of the peptides is a modified or unmodified fragment of the MAGA3 protein.

[0583] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 236016 to 236020.

[0584] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a MAGA4 protein. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 236016 to 236020.

[0585] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 236016 to 236020.

[0586] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 236016 to 236020. In some embodiments, one or more of the peptides is a modified or unmodified fragment of the MAGA4 protein.

[0587] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOS: 247059 to 247063.

[0588] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from the MAGC1 protein. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 247059 to 247063.

[0589] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 247059 to 247063.

[0590] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 247059 to 247063. In some embodiments, one or more of the peptides is a modified or unmodified fragment of the MAGC1 protein.

[0591] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 281350 to 281353.

[0592] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a MAGC3 protein. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 281350 to 281353.

[0593] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 281350 to 281353.

[0594] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 281350 to 281353. In some embodiments, one or more of the peptides is a modified or unmodified fragment of the MAGC3 protein.

[0595] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 369027 to 369031.

[0596] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from the SSX2 protein. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 369027 to 369031.

[0597] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 369027 to 369031.

[0598] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 369027 to 369031. In some embodiments, one or more of the peptides is a modified or unmodified fragment of the SSX2 protein.

[0599] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 342521 to 342525.

[0600] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a PRAME protein. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 342521 to 342525.

[0601] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 342521 to 342525.

[0602] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 342521 to 342525. In some embodiments, one or more of the peptides is a modified or unmodified fragment of the PRAME protein.

[0603] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 206663 to 206665.

[0604] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from the KKLC1 protein. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 206663 to 206665.

[0605] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 206663 to 206665.

[0606] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 206663 to 206665. In some embodiments, one or more of the peptides is a modified or unmodified fragment of the KKLC1 protein.

[0607] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 317360 to 317363.

[0608] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a PMEL protein, hi some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 317360 to 317363.

[0609] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 317360 to 317363.

[0610] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 317360 to 317363. In some embodiments, one or more of the peptides is a modified or unmodified fragment of the PMEL protein.

[0611] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 373348 to 373350.

[0612] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a TYRP1 protein. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 373348 to 373350.

[0613] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 373348 to 373350.

[0614] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 373348 to 373350. In some embodiments, one or more of the peptides is a modified or unmodified fragment of the TYRP1 protein.

[0615] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 392434 to 392437.

[0616] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a TYRP2 protein, hi some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 392434 to 392437.

[0617] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 392434 to 392437.

[0618] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 392434 to 392437. In some embodiments, one or more of the peptides is a modified or unmodified fragment of the TYRP2 protein.

[0619] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 305566 to 305570.

[0620] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from the MAR1 protein, hi some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 305566 to 305570.

[0621] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 305566 to 305570.

[0622] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 305566 to 305570. In some embodiments, one or more of the peptides is a modified or unmodified fragment of the MAR1 protein. Vaccines for autoimmune diseases

[0623] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOS: 34169 to 34204.

[0624] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from an INS protein. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 34169 to 34204.

[0625] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOS: 34169 to 34204.

[0626] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 34169 to 34204. In some embodiments, one or more of the peptides is a modified or unmodified fragment of the INS protein.

[0627] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 116478 to 116515.

[0628] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a MOG protein. In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 116478 to 116515.

[0629] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 116478 to 116515.

[0630] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 116478 to 116515. In some embodiments, one or more of the peptides is a modified or unmodified fragment of the MOG protein.

[0631] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 203517 to 203521.

[0632] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from an INS protein, hi some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 203517 to 203521.

[0633] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 203517 to 203521.

[0634] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 203517 to 203521. In some embodiments, one or more of the peptides is a modified or unmodified fragment of the INS protein.

[0635] In another aspect, the invention provides an immunogenic composition comprising a nucleic acid sequence encoding one or more amino acid sequences selected from the group consisting of SEQ ID NOs: 307670 to 307674.

[0636] In some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding one or more amino acid sequences derived from a MOG protein, hi some embodiments, the immunogenic composition comprises a nucleic acid sequence encoding two or more amino acid sequences selected from the group consisting of SEQ ID NOs: 307670 to 307674.

[0637] In another aspect, the present invention provides an immunogenic peptide composition comprising one or more peptides selected from the group consisting of SEQ ID NOs: 307670 to 307674.

[0638] In some embodiments, the immunogenic peptide composition comprises two or more peptides selected from the group consisting of SEQ ID NOs: 307670 to 307674. In some embodiments, one or more of the peptides is a modified or unmodified fragment of the MOG protein. [Brief description of the drawings]

[0639] The following figures illustrate exemplary embodiments of the present invention.

[0640] [Figure 1] FIG. 1 is a flow chart of the vaccine optimization method.

[0641] [Diagram 2]Figure 2 is a flow chart of the vaccine optimization method by seed set compression.

[0642] [Diagram 3] Figures 3A-3D show predicted population coverage of single-target MHC class I vaccines by vaccine size for the following mutations: BRAF V600E and BRAF V600M mutations (Figure 3A), EGFR A289V, EGFR G598V, and EGFR L858R (Figure 3B), KRAS G12D, KRAS G12V, KRAS G12R, KRAS G12C, KRAS G13D, KRAS G12A, and KRAS G12S (Figure 3C), and PIK3CA E542K, PIK3CA E545K, and PIK3CA H1047R (Figure 3D).

[0643] [Figure 4] Figures 4A-4C show predicted population coverage of single-target MHC class I vaccines by vaccine size for the following mutations: IDH1 R132H and IDH1 R132C (Figure 4A), NRAS Q61R, NRAS Q61K, and NRAS Q61L (Figure 4B), and TP53 R158L, TP53 R175H, TP53 R248Q, TP53 R273C, and TP53 R273H (Figure 4C).

[0644] [Diagram 5] Figures 5A-5D show predicted population coverage of single-target MHC class II vaccines by vaccine size for the following mutations: BRAF V600E and BRAF V600M (Figure 5A), EGFR A289V, EGFR G598V, and EGFR L858R (Figure 5B), KRAS G12D, KRAS G12V, KRAS G12R, KRAS G12C, KRAS G13D, KRAS G12A, and KRAS G12S (Figure 5C), and PIK3CA E542K, PIK3CA E545K, and PIK3CA H1047R (Figure 5D).

[0645] [Figure 6] Figures 6A-6C show predicted population coverage of single-target MHC class II vaccines by vaccine size for the following mutations: IDH1 R132H and IDH1 R132C (Figure 6A), NRAS Q61R, NRAS Q61K, and NRAS Q61L (Figure 6B), and TP53 R158L, TP53 R175H, TP53 R248Q, TP53 R273C, and TP53 R273H (Figure 6C).

[0646] [Figure 7] Figure 7 shows a table showing the respective probabilities of expression of various mutant protein targets in different cancers.

[0647] [Figure 8] FIG. 8 is a flow chart showing a method for optimizing a multi-target (combination) vaccine.

[0648] [Figure 9] Figures 9A-9D show predicted population coverage of multitargeted (mixed) MHC class I vaccines by vaccine size for pancreatic cancer (Figure 9A), colorectal cancer (Figure 9B), brain tumor (Figure 9C), and thyroid cancer (Figure 9D).

[0649] [Figure 10] Tables 10A-10B show the predicted population coverage of multitargeted (combined) MHC class I vaccines by vaccine size for bronchial and lung cancer (FIG. 10A) and skin cancer (FIG. 10B).

[0650] [Figure 11] Figures 11A-11D show predicted population coverage of multitargeted (mixed) MHC class II vaccines by vaccine size for pancreatic cancer (Figure 11A), colorectal cancer (Figure 11B), brain tumor (Figure 11C), and thyroid cancer (Figure 11D).

[0651] [Figure 12]12A-12B show predicted population coverage of multitargeted (mixed) MHC class II vaccines by vaccine size for bronchial and lung cancer (FIG. 12A) and skin cancer (FIG. 12B).

[0652] [Figure 13] Figure 13 is an example of a Python implementation of the MERGEMULTI function for the combination vaccine design procedure.

[0653] [Figure 14] FIG. 14 shows predicted peptide-HLA hits by vaccine size for the KRAS G12V vaccine against HLA diplotypes HLA-A02:03, HLA-A11:01, HLA-B55:02, HLA-B58:01, HLA-C03:02, and HLA-C03:03. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0654] In some embodiments, the present disclosure provides for peptide vaccines incorporating peptide sequences presented by major histocompatibility complex (MHC) molecules on cells to train the immune system to recognize cancer or pathogen disease cells. In some embodiments, the present disclosure provides for peptide vaccines incorporating peptide sequences presented by major histocompatibility complex (MHC) molecules on cells to induce therapeutic resistance in antigen-specific immunotherapy for autoimmune diseases (Alhadj Ali et al., 2017, Gibson et al., 2015). In some embodiments, the peptide vaccine is a composition of one or more peptides. In some embodiments, the peptide vaccine is an mRNA or DNA construct encoding one or more peptides, which is administered for expression in vivo.

[0655] Peptide presentation by MHC molecules is necessary, but not sufficient, for a peptide to be immunogenic and for the individual's T cells to recognize the resulting peptide-MHC complexes, leading to T cell activation, proliferation, and immunological memory. In some embodiments, ELISPOT (Slota et al., 2011) or multiplex identification of antigen-specific T cell receptors by a combination of immunoassays and immune receptor sequencing (MIRA) assays (Klinger et al., 2015) are used to score peptide presentation by MHC molecules (e.g., HLA alleles) (e.g., immunogenicity of peptides that require peptide binding) (e.g., measured as a peptide-HLA binding score). In some embodiments, experimental data from assays such as ELISPOT (Slota et al., 2011) or multiplex identification of antigen-specific T cell receptors by a combination of immunoassays and immune receptor sequencing (MIRA) assays (Klinger et al., 2015) can be used to generate a peptide-HLA immunogenicity index for peptides and HLA alleles in a particular experimental situation or individual. In some embodiments, experimental data from assays such as ELISPOT (Slota et al., 2011) or multiplexed identification of antigen-specific T cell receptors by a combination of immunoassays and immune receptor sequencing (MIRA) assays (Klinger et al., 2015) can be combined with machine learning-based predictions to score peptide presentation (e.g., binding affinity) by MHC molecules (e.g., HLA alleles) (e.g., measured as a peptide-HLA binding score) and to determine peptide-HLA immunogenicity indices. In some embodiments, MHCflurry or NetMHCpan (Reynisson et al., 2020) computational methods (as known in the art) are used to predict MHC class I presentation of peptides by HLA alleles (see Table 1). In some embodiments, NetMHCiiPan computational methods (Reynisson et al., 2020) are used to predict MHC class II presentation of peptides by HLA alleles (see Table 2).

[0656] In some embodiments, computational methods such as MHCflurry (Odonnell et al., 2018, Odonnell et al., 2020, incorporated herein by reference in its entirety), NetMHCpan (Reynisson et al., 2020, incorporated herein by reference in its entirety), and NetMHCiiPan (Reynisson et al., 2020) are used to predict MHC class I (MHCflurry, NetMHCpan) or class II (NetMHCiPan) presentation of peptides by HLA alleles. In other embodiments, other methods of determining peptide-HLA binding are used, such as those disclosed in International Patent Publication No. WO 2005 / 042698, incorporated herein by reference in its entirety. NetMHCpan-4.1 and NetMHcIiPan-4.0 use the NNAlign_MA algorithm (Alvarez et al., 2019, incorporated herein by reference in its entirety) for peptide-HLA binding prediction. NNAlign_MA is therefore based on the nNAlign (Nielsen et al., 2009, Nielsen et al., 2017, incorporated herein by reference in its entirety) neural network. NetMHCpan-4.1 (Reynisson et al., 2020) uses an NNAlign_MA network with at least 180 inputs (9 x 20 = 180 inputs) describing peptide sequences. Networks with 56 and 66 hidden neurons and two outputs have been used (Alvarez et al., 2019). Each network architecture (56 or 66 hidden neurons) was trained with five different random parameter initializations and five rounds of cross-validation, resulting in a total of 50 distinct trained networks (2 architectures x 5 initializations x 5 cross-validations).These 50 trained networks are used as an ensemble of 25 networks with at least 10,800 parameters (180 inputs x 56 neurons) and 25 networks with at least 11,880 parameters (180 inputs x 66 neurons). The ensemble of 50 networks in NetMHCpan-4.1 therefore consists of a minimum of 567,000 parameters that should be evaluated in at least 567,000 arithmetic operations to calculate peptide-MHC binding. NetMHCIIpan-4.1 (Reynisson et al., 2020) uses NNAlign_MA networks with at least 180 inputs (9 x 20 = 180 inputs) describing peptide sequences. Networks with 2, 10, 20, 40 and 60 hidden neurons and two outputs are used (Alvarez et al., 2019). Each network architecture (2, 10, 20, 40 or 60 hidden neurons) is trained with 10 different random parameter initializations and 5 rounds of cross-validation, resulting in a total of 250 distinct trained networks (5 architectures x 10 initializations x 5 rounds of cross-validation). These 250 trained networks are used in ensembles of 50 networks with at least 360 parameters (180 inputs x 2 neurons), 50 networks with at least 1800 parameters (180 inputs x 10 neurons), 50 networks with at least 3600 parameters (180 inputs x 20 neurons), 50 networks with at least 7200 parameters (180 inputs x 40 neurons), and 50 networks with more than 10800 parameters (180 inputs x 60 neurons).Thus, the ensemble of 250 networks in NetMHCIIpan-4.0 consists of a minimum of 1,118,000 parameters that must be evaluated in at least 1,188,00 arithmetic operations to calculate peptide-MHC binding.

[0657] In some embodiments, the computational methods used to predict either MHC class I (e.g., MHCflurry, NetMHCpan) or class II (e.g., NetMHCIIPan) peptide-HLA binding scores or peptide-HLA immunogenicity metrics are based on data from experimental mass spectrometry observations of peptides bound by MHC molecules. In some embodiments, the computational methods used to predict either MHC class I (e.g., MHCflurry, NetMHCpan) or class II (e.g., NetMHCIIPan) peptide-HLA binding scores or peptide-HLA immunogenicity metrics are based on data from experimental observations of peptide-MHC binding affinity. In some embodiments, experimental observations of peptide-MHC binding affinity or immunogenicity, including mass spectrometry measurements of peptide-HLA binding and measurements of T cell activation, can be found in databases such as the Immune Epitope Database (IEDB) (Vita et al., 2018). MHCflurry 2.0 output (Odonnell et al., 2020, incorporated herein by reference in its entirety) is based on 493,473 mass spectrometry measurements of peptide-HLA binding and 219,596 affinity measurements of peptide-HLA binding. NetMHCpan-4.1 output (Reynisson et al., 2020) is based on 665,492 mass spectrometry measurements of peptide-HLA binding and 52,402 affinity measurements of peptide-HLA binding. NetMHCIIpan-4.0 output (Reynisson et al., 2020) is based on 381,066 mass spectrometry measurements of peptide-HLA binding and 44,861 affinity measurements of peptide-HLA binding.

[0658] Presentation of peptides is by MHC molecules and occurs when the peptide binds in the groove of the MHC molecule and is transported to the cell surface where it is recognized by the T cell receptor. Target peptide refers to a foreign or self-peptide. In some embodiments, a peptide that is part of the normal proteome in a healthy individual is a self-peptide, and a peptide that is not part of the normal proteome is a foreign peptide. In some embodiments, a target peptide can be part of the normal proteome that shows abnormal expression (e.g., a cancer testis antigen such as NY-ESO-1). Foreign peptides are generated by mutations of normal self-proteins of tumor cells that generate epitopes called neoantigens or by pathogenic infections. In some embodiments, a neoantigen is any subsequence of a human protein that contains one or more altered amino acids or protein modifications that do not appear in healthy individuals. Thus, in the present disclosure, a foreign peptide refers to an amino acid sequence that encodes a fragment of a target protein / peptide (or a full-length protein / peptide), i.e., a target protein / peptide consisting of a neoantigenic protein, a pathogen proteome, or other undesirable protein that is non-self and predicted to be bound and displayed by HLA alleles.

[0659] Protein genes frequently mutated in cancer identified by UniProt ID include AKT1_HUMAN, BRAF_HUMAN, EGFR_HUMAN, GTF2I_HUMAN, RASH_HUMAN (also known as HRAS), IDHC_HUMAN (also known as IDH1), RASK_HUMAN (also known as KRAS), RASN_HUMAN (also known as NRAS), PIK3CA_HUMAN, PTEN_HUMAN, and P53_HUMAN (also known as TP53). Here, missense mutations in proteins are described with the single-letter amino acid code of the wild-type amino acid, the position of the amino acid mutation, and the single-letter amino acid code present in the mutated protein. For example, KRAS G12D is a mutation in the KRAS protein where position 12 is changed from glycine to aspartic acid (G12D). A protein may contain multiple mutations at various positions. For brevity, genes may be referred to here without the suffix "_HUMAN".

[0660] KRAS gene mutations are the most common mutated oncogene in cancer, but have been extremely difficult to treat using small molecule therapeutics. The KRAS protein is part of a signaling pathway that controls cell proliferation, and point mutations in this protein can lead to constitutive pathway activation and uncontrolled cell proliferation. Single amino acid KRAS mutations result in only minor changes in the protein structure, making it difficult to design small molecule drugs that recognize the specific binding pocket of the mutant and inactivate KRAS signaling. Oncogenic mutations in KRAS include glycine to aspartic acid (G12D), glycine to valine (G12V), glycine to arginine (G12R), or glycine to cysteine ​​mutations at position 12 (G12C), or glycine to aspartic acid mutation at position 13 (G13D). The corresponding exogenous peptides include these mutations. KRAS is a member of the RAS family of genes, which includes HRAS and NRAS. KRAS, HRAS, and NRAS have identical sequences from residue 1 to residue 86. Thus, all vaccines and peptide sequences described herein for a mutation in one RAS family member can be used for the same mutation in any other RAS family member (eg, a KRAS G12D vaccine is also a vaccine for HRAS G12D).

[0661] Certain self-proteins, such as cancer-testis antigens, are present at abnormally high levels on cancer cells and therefore may be targets for vaccines that induce intolerance of T cell responses to cells presenting peptides derived from these self-proteins on MHC molecules. Examples of these cancer-associated proteins by UniProt ID include CTG1B_HUMAN (also known as NY-ESO-1), MAGA1_HUMAN, MAGA3_HUMAN, MAGA4_HUMAN, MAGC1_HUMAN, MAGC3_HUMAN, SSX2_HUMAN, PRAME_HUMAN, KKLC1_HUMAN (also known as CT83), PMEL_HUMAN (also known as gp100), TYRP1_HUMAN (also known as gp75), TYRP2_HUMAN (also known as DCT), and MAR1_HUMAN.

[0662] Autoimmune diseases are caused by an immune system that has lost self-tolerance to self-proteins. Such loss of self-tolerance is involved in autoimmune diseases such as diabetes, multiple sclerosis, and autoimmune encephalomyelitis. Induction of tolerance to autoimmune-associated self-peptides can be achieved by antigen-specific tolerization using delivery of vaccine antigens through tolerization protocols. An example of a protocol for inducing tolerance with lipid nanoparticle (LNP) encapsulated (mRNA-LNP) vaccines is described by Krienke, et al., 2021, incorporated herein by reference in its entirety. Examples of autoimmune disease-associated proteins include the UniProt IDs INS_HUMAN (also known as insulin) and MOG_HUMAN (also known as myelin-oligodendrocyte glycoprotein). Patients with diabetes may have a loss of tolerance to INS_HUMAN, and patients with multiple sclerosis or autoimmune encephalomyelitis may have a loss of tolerance to MOG_HUMAN, respectively.

[0663] A challenge in designing peptide vaccines is that the diversity of human MHC alleles (HLA alleles) means that each allele has a specific preference for the peptide sequence displayed. Human leukocyte antigen (HLA) loci within the MHC encode HLA class I and class II molecules. There are three classical class I loci (HLA-A, HLA-B, and HLA-C) and three loci that encode class II molecules (HLA-DR, HLA-DQ, and HLA-DP). An individual's HLA type represents the alleles located at each of these loci. Peptides with a length of about 8 to about 11 residues can bind to HLA class I (or MHC class I) molecules, whereas peptides with a length of about 13 to about 25 residues bind to HLA class II (or MHC class II) molecules (Rist et al., 2013; Chicz et al., 1992). Human populations from different geographic regions have different frequencies of HLA alleles, and these populations exhibit linkage disequilibrium between HLA loci, resulting in population-specific haplotype frequencies. In some embodiments, methods for producing effective vaccines are disclosed that include considering the HLA allele frequencies in a target population and considering the linkage disequilibrium between HLA genes to achieve a set of peptides that are likely to be reliably displayed.

[0664] The present disclosure provides compositions, systems, and methods for designing vaccines that confer immunity to single or multiple targets. In some embodiments, the targets are neoantigen protein sequences, pathogen proteomes, or other non-self, undesired protein sequences that are expected to be bound and presented by HLA molecules (also referred to herein as HLA alleles). When a target is present in an individual, multiple peptide sequences may be presented by different HLA alleles. In some embodiments, it may be desirable to create a vaccine that includes selected self-peptides, and therefore, these selected self-peptides are considered to be target peptides for this purpose.

[0665] The term peptide-HLA binding is defined as the binding of a peptide to an HLA allele, which can be either computationally predicted, experimentally observed, or computationally predicted using experimental observations. The peptide-HLA binding index can be expressed as affinity, percentile rank, binary at a given threshold, probability, or other index known in the art. The term peptide-HLA immunogenicity index is defined as the activation that occurs in T cells based on the recognition of a peptide when bound to an HLA allele. The term peptide-HLA immunogenicity score is another name for peptide-HLA immunogenicity index, and these terms have the same meaning. The peptide-HLA immunogenicity index can vary from person to person, and the peptide-HLA immunogenicity index can be expressed as a probability, a binary index, or other index related to the likelihood that a peptide-HLA combination is immunogenic. In some embodiments, peptide-HLA immunogenicity is defined as the induction of immune tolerance based on the recognition of a peptide when bound to an HLA allele. The peptide-HLA immunogenicity index can be computationally predicted, experimentally observed, or computationally predicted using experimental observations. In some embodiments, the peptide-HLA immunogenicity index is based solely on peptide-HLA binding, since peptide-HLA immunogenicity requires peptide-HLA binding. In some embodiments, peptide-HLA immunogenicity data or computational predictions of peptide-HLA immunogenicity can be included and combined in the score of peptide presentation in the methods disclosed herein. One way to combine scores is to use immunogenicity data of peptides analyzed for immunogenicity in diseased or vaccinated individuals, select HLA alleles predicted by computational methods to be most likely to be presented, and assign peptides to HLA alleles presented in the individual. For peptides that have not been experimentally analyzed, computational predictions of presentation can be used. In some embodiments, various computational methods of predicting peptide-HLA immunogenicity or peptide-HLA binding can be combined (Liu et al., 2020b).For a particular set of peptides and HLA alleles, the term peptide-HLA hits refers to the number of unique combinations of peptides and HLA alleles that show peptide-HLA immunogenicity or binding at a given threshold. For example, a peptide-HLA hit of 2 means that one of the following is expected: one peptide binds to two different HLA alleles (or causes T cell activation), or two peptides bind to two different HLA alleles (or causes T cell activation), or two peptides bind to the same HLA allele (or causes T cell activation). For a particular set of peptides and HLA frequencies, HLA haplotype frequencies, or HLA diplotype frequencies, the expected number of peptide-HLA hits is the average number of peptide-HLA hits for each set representing an individual, corrected for frequency of occurrence.

[0666] Because immunogenicity may vary between individuals, one way to increase the efficacy of a vaccine is to use a diverse set of target peptides (e.g., at least two peptides) to increase the likelihood that some subset of them will be immunogenic in a given individual. Previous studies using mouse models have shown that most MHC-displayed peptides are immunogenic, but as Croft et al. (2019) explain, immunogenicity varies between individuals. In some embodiments, experimental peptide-HLA immunogenicity data is used to determine which target peptides and their modifications will be effective immunogens in a vaccine.

[0667] Considerations for peptide vaccine design are reviewed in Liu et al., Cell Systems 11, Issue 2, p. 131-146 (Liu et al., 2020 and Liu et al., 2020b), and in U.S. Pat. Nos. 11,058,751 and 11,161,892, which are incorporated by reference in their entireties.

[0668] Certain target peptides may not bind with high affinity to a wide range of HLA molecules. To increase binding of target peptides to HLA molecules, their amino acid composition can be altered to change one or more anchor residues or other residues. In some embodiments, to increase the immunogenicity of the target peptide when presented by HLA molecules, the amino acid composition of the target peptide can be altered to change one or more residues. Anchor residues are amino acids that interact with HLA molecules and have the greatest effect on the affinity of the peptide to HLA molecules. Peptides with one or more altered amino acid residues are called heteroclitic peptides. In some embodiments, heteroclitic peptides include target peptides with residue modifications at anchor positions. In some embodiments, heteroclitic peptides include target peptides with residue modifications at non-anchor positions. In some embodiments, heteroclitic peptides include target peptides with residue modifications that include non-natural amino acids and amino acid derivatives. Modifications to generate heteroclitic peptides can improve binding of the peptide to both MHC class I and MHC class II molecules, and the required modifications can be specific to both the peptide and the MHC class. Peptide anchor residues face the MHC molecule groove and are therefore less likely to be recognized by T cell receptors than other peptide residues. Thus, heteroclitic peptides with modified anchor residues have been observed to elicit T cell responses and stimulated T cells also respond to unmodified peptides. The use of heteroclitic peptides in vaccines has been observed to improve vaccine efficacy (Zirlik et al., 2006). In some embodiments, the immunogenicity of heteroclitic peptides is experimentally determined to identify their ability to activate T cells that also recognize the corresponding base (also called seed) peptide of the heteroclitic peptide, as known in the art (Houghton et al., 2007).In some embodiments, these assays for immunogenicity and cross-reactivity of heteroclitic peptides are performed when the heteroclitic peptide is presented by a particular HLA allele. Peptide vaccines that induce immunity against one or more targets

[0669] In some embodiments, a method is provided for tailoring peptide vaccines with a single vaccine design against one or more targets. In some embodiments, the single target is a foreign protein with a specific mutation (e.g., KRAS G12D). In some embodiments, the single target is a self protein (e.g., a protein overexpressed in tumor cells, such as a cancer / testis antigen). In some embodiments, the single target is a pathogen protein (e.g., a protein contained in a viral proteome). In some embodiments, multiple targets (e.g., both KRAS G12D and KRAS G13D) can be used.

[0670] In some embodiments, the method involves extracting peptides and constructing a candidate set from all target proteome sequences (e.g., the entire KRAS G12D protein) as described by Liu et al. (2020).

[0671] Figures 1 and 2 show a flow chart of an example vaccine design method that can be used for MHC class I or MHC class II vaccine design. The candidate peptide set (see Figures 1 and 2) is composed of target peptides extracted by windowing the input protein sequence. In some embodiments, the extracted target peptides are about 8 to about 10 amino acid residues long (e.g., for MHC class I binding) (Rist et al., 2013). In some embodiments, the extracted target peptides presented by MHC class I molecules are longer than 10 amino acid residues, such as 11 residues (Trolle et al., 2016). In some embodiments, the extracted target peptides are about 13 to about 25 amino acid residues long (e.g., for class II binding) (Chicz et al., 1992). In some embodiments, sliding windows of various size ranges described herein are used across the proteome. In some embodiments, other target peptide lengths for MHC class I and class II sliding windows can be utilized. In some embodiments, computational prediction of proteasomal cleavage is used to filter or select peptides in the candidate set. One computational method for predicting proteasomal cleavage is described by Nielsen et al. (2005). In some embodiments, peptides can be filtered using peptide mutation rates, glycosylation, cleavage sites, or other criteria, as described by Liu et al. (2020). In some embodiments, peptides can be filtered based on evolutionary sequence variation above a predefined threshold. Evolutionary sequence variation can be calculated with respect to other species, other pathogens, other pathogen strains, or other related organisms. In some embodiments, the first peptide set is the candidate set.

[0672] As shown in Figures 1-2, in some embodiments, the next step of the method includes scoring the target peptides in the candidate set for peptide-HLA binding to all possible HLA alleles as described by Liu et al. (2020) and Liu et al. (2020b). In some embodiments, the first peptide set is the candidate set after scoring the target peptides. Scoring can be performed for human HLA molecules, mouse H-2 molecules, porcine SLA molecules, or MHC molecules of any species for which a predictive algorithm is available or can be developed. Thus, vaccines can be designed for non-human species in this manner. The scoring matrix includes the affinity of the target peptide for the HLA allele in nanomolar, eluted ligand, presentation, and other scores that can be expressed as percentile rank or other indices. The candidate set can be further filtered to exclude peptides whose predicted binding core does not contain the target residue of the specific pathogenic or neo-antigen of interest or whose predicted binding core contains the target residue at the anchor position. The candidate set can also be filtered for target peptides of a certain length, for example, length 9 for MHC class I. In some embodiments, the scoring of target peptides is performed using experimental data or a combination of experimental data and computational prediction methods. If a computational model is not available to perform peptide-HLA binding prediction for a particular (peptide, HLA) pair, the binding value for such a pair can be estimated using other techniques, including the mean, median, minimum, or maximum immunogenicity taken for the supported pairs, a fixed value (such as a value indicating no binding), or a prediction function of the most similar (peptide, HLA) pair available in the scoring model.

[0673] In some embodiments, a base set (also referred to herein as a seed set) is constructed by selecting peptides from a candidate set scored using individual peptide-HLA binding or immunogenicity criteria (e.g., a first peptide set) (Figure 1). In some embodiments, a given peptide has multiple peptide-HLA scores, so that one with the best affinity or highest immunogenicity (e.g., predicted to bind most strongly or be most active for a given HLA allele) can be selected based on the peptide-HLA binding score or peptide-HLA immunogenicity index. The criteria used in scoring peptide-HLA binding during the scoring procedure can correspond to different goals in the base set selection and vaccine design phase. For example, a target peptide with a peptide-HLA binding affinity of 500 nM may be presented by diseased individuals, but less frequently than a target peptide with a peptide-HLA binding affinity of 50 nM. In the vaccine combination design stage, a more stringent affinity criterion, such as 50 nM (e.g., in the case of selecting the third peptide set for the vaccine targeting shown in Figures 1 and 2), can be used to increase the likelihood that the vaccine peptide will be found and presented by the HLA molecule. In some embodiments, a relatively loose threshold of peptide-HLA immunogenicity or peptide-HLA binding (e.g., less than about 1000 nM or less than about 500 nM) is used as a first threshold to filter the candidate peptide-HLA scores (first peptide scoring and score filtering step in Figures 1 and 2), and a relatively stringent second threshold (e.g., less than about 50 nM) is used to filter the expanded set of peptide-HLA scores for a particular HLA allele (second peptide filtering and scoring step in Figures 1 and 2). In some embodiments, a particular peptide-HLA score is not used for a modified peptide for a given HLA for vaccine design if the unmodified corresponding peptide does not exceed the first, less restrictive threshold.This filtering of peptide-HLA scores is based on the observation that peptides that are not sufficiently immunogenic for inclusion in a vaccine are antigenic (meeting the first filtering threshold) and therefore may be recognized by T cell clonotypes expanded by the vaccine. Peptides are antigenic when recognized by T cell receptors, eliciting responses such as CD8+ T cell cytotoxicity and CD4+ cell activation. Derivatives of antigenic peptides may be potently immunogenic when included in a vaccine, thus activating and expanding T cells that recognize the antigenic peptide. Expansion of T cells that recognize unmodified antigenic peptides can result in an immune response that contributes to disease control. In some embodiments, peptides are scored for potential inclusion in a third peptide set (vaccine versus Figures 1 and 2) that has a peptide-HLA binding affinity of less than about 500 nM. In some embodiments, peptides that have a peptide-HLA binding affinity of less than about 1000 nM for at least one HLA allele are selected for the base set. Alternatively, peptide-HLA immunogenicity predictions can be used to determine the suitability of target peptides for inclusion in the base set. In some embodiments, experimental observations of the immunogenicity of peptides in the context of presentation by HLA alleles, or binding of peptides to HLA alleles, can be used to score peptide binding to HLA alleles or peptide-HLA immunogenicity.

[0674] In some embodiments, experimental observations of peptide presentation by specific HLA alleles in tumor cells can be used to score peptides for peptide-HLA binding or peptide-HLA immunogenicity. In some embodiments, experimental observations of tumor cell peptide presentation by specific HLA alleles can be used to score peptides for peptide-HLA binding or peptide-HLA immunogenicity for that HLA allele. In some embodiments, experimental observations of tumor cell peptide presentation can be used to score peptides for peptide-HLA binding or peptide-HLA immunogenicity using HLA alleles for a particular observed peptide, selected from HLA alleles present in the tumor that meet a predicted threshold for peptide-HLA binding or immunogenicity. In some embodiments, mass spectrometry is used to experimentally determine peptide presentation by tumor cells, as described by Bear et al. (2021) or Wang et al. (2019), and these data are used to evaluate peptide-HLA binding or peptide-HLA immunogenicity. In some embodiments, the presentation of peptides by tumor cells is experimentally determined using mass spectrometry, and these experimental data are used to determine whether the vaccine contains base set (seed set) peptides for one or more HLA alleles. In some embodiments, the presentation of peptides by tumor cells is experimentally determined using mass spectrometry, and these experimental data are used to exclude peptide-HLA binding scores or peptide-HLA immunogenicity scores when the peptide is not observed by mass spectrometry to be presented by an HLA allele. In some embodiments, the presentation of peptides by tumor cells is experimentally determined using mass spectrometry, and these experimental data are used to determine whether the vaccine contains base set (seed set) peptides for one or more HLA alleles.In some embodiments, mass spectrometry is used to experimentally determine the presentation of peptides by tumor cells of an individual, and these experimental data are used to exclude peptide-HLA binding scores or peptide-HLA immunogenicity scores for peptides where the peptide is not observed by mass spectrometry to be presented by an HLA allele. In some embodiments, computational predictions of the immunogenicity of peptides in the context of presentation by HLA alleles can be used for scoring, such as those in Ogishi et al. (2019) or Bulik-Sullivan et al. (2019).

[0675] In some embodiments, if the wild-type peptide corresponding to the first peptide (e.g., the naturally occurring unmutated form of the peptide, or the respective species peptide within a defined sequence edit distance) has a peptide-HLA score or peptide-HLA immunogenicity score for the same HLA allele within a defined threshold, the peptide-HLA score or peptide-HLA immunogenicity score for the first peptide in the base set (seed set) of a given HLA allele is removed and not considered in the vaccine design. The threshold can be based on the difference between the scores of the first peptide and the wild-type peptide, the ratio of the scores of the first peptide and the wild-type peptide, the score of the wild-type peptide, or other indicators. The defined threshold can be greater or less than a specified value. In some embodiments, the threshold is defined such that the wild-type peptide is not expected to be presented. In some embodiments, when the peptide-HLA score or peptide-HLA immunogenicity score for a first peptide is removed during vaccine design, the peptide-HLA scores or peptide-HLA immunogenicity scores of all derivatives of the same HLA allele (e.g., heteroclitic peptide derivatives) are also removed and are not considered during vaccine design.

[0676] In some embodiments, the method further includes running the OptiVax-Robust algorithm, as described by Liu et al. (2020), to construct a base set of target peptides (also referred to herein as a seed set) using the population HLA haplotype frequencies on the scored candidate set (Figure 2). In some embodiments, the HLA diplotype frequencies can be provided to OptiVax. OptiVax-Robust includes an algorithm to eliminate peptide redundancy resulting from a sliding window approach at various window sizes, although other means of removing redundancy can also be used to enforce a minimum edit distance constraint between target peptides in the candidate set. The size of the seed set is determined by the point of diminishing returns in population coverage as a function of the number of target peptides in the seed set. Other criteria can also be used, such as a minimum number of vaccine target peptides, a maximum number of vaccine target peptides, and a desired expected population coverage. In some embodiments, the predetermined population coverage is less than about 0.4, about 0.4-0.5, about 0.5-0.6, about 0.6-0.7, about 0.7-0.8, about 0.8-0.9, or greater than about 0.9. Another possible criterion is the minimum number of peptide-HLA binding hits expected for each individual. In another embodiment, the method further includes running the OptiVax-Unlinked algorithm instead of OptiVax-Robust, as described in Liu et al. (2020).

[0677] The OptiVax-Robust method uses binary predictions of peptide-HLA immunogenicity, and these binary predictions can be generated as described in Liu et al. (2020b). The OptiVax-Unlinked method uses the probability of binding of the target peptide to an HLA allele, and these can be generated as described in Liu et al. (2020). In some embodiments, OptiVax Unlinked and EvalVax Unlinked are used with the probability of peptide-HLA immunogenicity being observed. For the purposes described herein, either method can be used, so the term "OptiVax" refers to either the Robust or Unlinked methods. In some embodiments, the probability of peptide-HLA immunogenicity observed in experimental assays can be used as the probability of peptide-HLA binding in EvalVax Unlinked and OptiVax Unlinked. In some embodiments, the HLA haplotype or HLA allele frequencies of the population provided to OptiVax for vaccine design are representative of the world's population. In another embodiment, the HLA haplotype or HLA allele frequency of the population provided to OptiVax for vaccine design is specific to a geographic region. In another embodiment, the HLA haplotype or HLA allele frequency of the population provided to OptiVax for vaccine design is specific to a bloodline. In another embodiment, the HLA haplotype or HLA allele frequency of the population provided to OptiVax for vaccine design is specific to a race. In another embodiment, the HLA haplotype or HLA allele frequency of the population provided to OptiVax for vaccine design is specific to individuals with risk factors such as genetic indicators of risk, age, exposure to chemicals, alcohol intake, chronic inflammation, diet, hormones, immunosuppression, infectious agents, obesity, radiation, sunlight, or smoking. In another embodiment, the HLA haplotype or HLA allele frequency of the population provided to OptiVax for vaccine design is specific to individuals with certain HLA alleles.In another embodiment, the HLA diplotypes provided to OptiVax for vaccine design describe a single individual and are used to design a personalized vaccine.

[0678] In some embodiments, the base set (or seed set) of target peptides resulting from the application of OptiVax to a candidate set of target peptides (e.g., first peptide set) represents a set of unmodified target peptides that may compact vaccine design (seed set in FIG. 2). The peptide bases are the target peptides included in the base or seed peptide set (e.g., first peptide set). In some embodiments, the seed set (e.g., first peptide set) is based on filtering candidate peptide scores by predicted or observed affinity or immunogenicity to HLA molecules (seed set in FIG. 1). However, in order to improve the presentation of target peptides in the widest possible range of HLA haplotypes, some embodiments include modifications of the seed (or base) set. In some embodiments, experimental assays can be used to confirm that modified seed (or base) peptides activate T cells that also recognize the base / seed peptide.

[0679] For a particular target peptide, the selection of optimal anchor residues may depend on the HLA alleles that bind and present the target peptide and the class of HLA alleles (MHC class I or class II). The seed peptide set (e.g., first peptide set) can be expanded by including anchor residue-modified peptides of either MHC class I or II peptides (Figure 1-2). Thus, one aspect of vaccine design is to consider how to select a limited set of heteroclitic peptides derived from the same target peptide for inclusion in the vaccine, knowing that different heteroclitic peptides may have different and overlapping population coverage.

[0680] In some embodiments, all possible anchor modifications are considered for each base set of the target peptide. Peptides bound to MHC class I molecules typically have two anchor residues, typically positions 2 and 9 for 9-mer peptides. In some embodiments, the anchors for 8-mers, 10-mers, and 11-mers are at positions 2 and n, where n is the last position (8, 10, and 11, respectively). For MHC class I molecules, the last position, n, is referred to herein as the carboxyl-terminal "C" position. In some embodiments, 20 possible amino acids are tried at each anchor position to select the best heteroclitic peptide. Thus, for MHC class I binding, 400 (i.e., 20 amino acids x 2 positions = 20) are tried for each base target peptide. 2 ) minus one heteroclitic peptide. Peptides bound to MHC class II molecules typically have four anchor residues at positions 1, 4, 6, and 9 of the 9-mer binding core. Thus, for MHC class I binding, 160,000 (i.e., 20 amino acids x 4 positions = 20 4 ) minus one heteroclitic peptide is generated. In some embodiments, two or more (MHC class I) or four (MHC class II) positions are considered anchors. Other methods, including Bayesian optimization, can be used to select optimal anchor residues and generate heteroclitic peptides from each seed (or base) set peptide. Other methods for selecting optimal anchor residues are presented in "Machine learning optimization of peptides for presentation by class II MHCs" by Dai et al. (2020), which is incorporated herein in its entirety. In some embodiments, the anchor positions are determined by the HLA alleles that present the peptides, so that the set of heteroclitic peptides includes all possible anchor modifications for each set of HLA-specific anchor positions.

[0681] In some embodiments, new peptide sequences (e.g., MHC class I or class II) with all possible anchor residue modifications are created for all target peptides in the base / seed set, resulting in a new heteroclitic base set (expanded set in Figure 1-2) that includes all modifications. In some embodiments, if one or more of the anchor residue positions of the peptide contain a substitution mutation that distinguishes the peptide from a self-peptide, the anchor residue modifications of the peptide are not included in the heteroclitic base set. In some embodiments, the anchor residue modifications of the base / seed peptide are only included in the heteroclitic base set at peptide positions that do not contain a substitution mutation that distinguishes the base / seed peptide from a self-peptide. In some embodiments, the anchor residue modifications of the peptide are not included in the heteroclitic base set if one or more mutations of the peptide do not occur between its adjacent anchor residue pairs. In some embodiments, new peptide sequences (e.g., MHC class I or class II) with anchor residue modifications are created for all target peptides in the base / seed set, resulting in a new heteroclitic base set (expanded set in Figure 1-2) that includes all modifications. In some embodiments, the anchor residue positions used to modify the peptides are selected from the anchor residue positions determined by the HLA alleles considered during vaccine evaluation. In some embodiments, the heteroclitic base set (expanded set in Figure 1-2) also includes the original seed (or base) set (seed peptide set in Figure 1-2). In some embodiments, the heteroclitic base set includes amino acid substitutions at non-anchor residues. In some embodiments, base peptide residues are modified to improve therapeutic efficacy by altering binding to T cell receptors (Candia et al., 2016). In some embodiments, the heteroclitic base set includes amino acid substitutions of non-natural amino acid analogs.The heteroclitic base sets are scored for HLA affinity, peptide-HLA immunogenicity, or other indicators described herein (a second peptide scoring and score filtering process shown in Figures 1-2). In some embodiments, the scoring predictions may be further updated for heteroclitic peptide and HLA allele pairs, eliminating pairs in which the heteroclitic peptide has a seed (or base) peptide that is not predicted to be presented by the HLA allele at a particular threshold of peptide-HLA binding score or particular peptide-HLA immunogenicity indicator. In some embodiments, the peptide-HLA scores may also be filtered to ensure that the predicted binding core of a heteroclitic peptide presented by a particular HLA allele is exactly aligned with the binding core of the respective seed (or base) set target peptide set for that HLA allele. In some embodiments, the scoring predictions are filtered for the HLA allele to ensure that the heteroclitic peptides considered for that HLA allele are modified only at anchor positions determined by that HLA allele. Scoring provides an indicator of peptide-HLA immunogenicity for the peptide and HLA allele. The index may be either binary, a probability of immunogenicity, or other immunogenicity index such as peptide-HLA affinity or percent rank, and may be based on computational prediction, experimental observation, or a combination of both computational prediction and experimental observation. In some embodiments, the probability of peptide-HLA immunogenicity is utilized by OptiVax-unbound. In some embodiments, heteroclitic peptides are included in experimental assays such as MIRA (Klinger et al., 2015) or ELISPOT to determine the peptide-HLA immunogenicity index for a particular HLA allele. In some embodiments, using the method of Liu et al. (2020b), MIRA data for heteroclitic peptides can be incorporated into a model of peptide-HLA immunogenicity.In some embodiments, the peptide-HLA immunogenicity index of a heteroclitic peptide is experimentally determined and its ability to activate T cells that also recognize the seed (or base) peptide corresponding to the heteroclitic peptide is confirmed by methods known in the art to qualify the heteroclitic peptide for inclusion in a vaccine (e.g., Houghton et al., 2007). In some embodiments, these assays of immunogenicity and cross-reactivity of heteroclitic peptides are performed when the heteroclitic peptide is presented by a particular HLA allele.

[0682] In some embodiments, experimental observations of presentation of heteroclitic peptides by specific HLA alleles in cells can be used to score peptides for peptide-HLA binding or peptide-HLA immunogenicity. In some embodiments, mass spectrometry is used to experimentally determine presentation of heteroclitic peptides by cells, as described by Bear et al. (2021) or Wang et al. (2019), and these data are used to evaluate peptide-HLA binding or peptide-HLA immunogenicity. In some embodiments, mass spectrometry is used to experimentally determine presentation of heteroclitic peptides by cells, and these experimental data are used to qualify the inclusion of the heteroclitic peptide in a vaccine. In some embodiments, mass spectrometry is used to experimentally determine presentation of peptides by tumor cells, and these experimental data are used to exclude peptide-HLA binding scores or peptide-HLA immunogenicity scores when the peptide is not observed by mass spectrometry to be displayed by an HLA allele. In some embodiments, mass spectrometry is used to experimentally determine presentation of a heteroclitic peptide by cells with HLA alleles found in an individual, and these experimental data are used to qualify the heteroclitic peptide for inclusion in a vaccine for the individual. In some embodiments, mass spectrometry is used to experimentally determine presentation of a peptide by tumor cells, and these experimental data are used to exclude a peptide-HLA binding score or peptide-HLA immunogenicity score if the peptide is not observed by mass spectrometry to be presented by an HLA allele. In some embodiments, computational predictions of the immunogenicity of a peptide in the context of presentation by an HLA allele can be used for scoring in the manner of Ogishi et al. (2019) or Bulik-Sullivan et al. (2019).

[0683] In some embodiments, peptides in the heteroclitic base set are removed if (1) one of the anchor positions of the HLA allele corresponds to a position of a mutation in the base / seed peptide that results from a mutation that distinguishes the base / seed peptide from a self-peptide, and (2) the peptide-HLA binding or peptide-HLA immunogenicity of the self-peptide is stronger than a certain threshold of self-peptide binding or immunogenicity. This removes peptides in the heteroclitic base set that may cross-react with the self-peptide as a result of sharing TCR-facing residues with the self-peptide. In some embodiments, the threshold of self-peptide binding is about 500 nM to 1000 nM.

[0684] In some embodiments, redundant peptides in the heteroclitic base set are removed. In some embodiments, the redundant peptide is a first heteroclitic peptide that has a peptide-HLA immunogenicity score or peptide-HLA binding score that is less immunogenic to all HLAs than a second heteroclitic peptide in the heteroclitic base set, but in which both the first and second heteroclitic peptides are derived from the same base (or seed) peptide. In some embodiments, the redundancy of peptides is determined only by comparing the peptide-HLA immunogenicity scores or peptide-HLA binding scores of HLA alleles if the peptide-HLA immunogenicity scores or peptide-HLA binding scores of both peptides of the HLA alleles are more immunogenic than a predetermined threshold (e.g., 50 nM for binding). In some embodiments, a redundant peptide is a first heteroclitic peptide having an average peptide-HLA immunogenicity score or peptide-HLA binding score that is less immunogenic than the average peptide-HLA immunogenicity score or peptide-HLA binding score of a second heteroclitic peptide in the heteroclitic base set, but where both the first and second heteroclitic peptides in the heteroclitic base set are derived from the same base (or seed) peptide, and the peptide-HLA immunogenicity scores or peptide-HLA binding scores of both peptides for an HLA allele are more immunogenic than a predetermined threshold (e.g., 50 nM for binding), then an average score for the HLA allele is calculated. In some embodiments, the redundant peptide is a first heteroclitic peptide having a corrected peptide-HLA immunogenicity score or peptide-HLA binding score that is less immunogenic than the corrected peptide-HLA immunogenicity score or peptide-HLA binding score of a second heteroclitic peptide in a heteroclitic base set, where both the first and second heteroclitic peptides are derived from the same base (or seed) peptide, and the correction is determined by the HLA allele frequency in the human population.A corrected score is calculated if the peptide-HLA immunogenicity score or peptide-HLA binding score of both peptides for an HLA allele is higher than a predefined threshold of immunogenicity (e.g., 50 nM for binding).

[0685] In some embodiments, the next step is to create a second peptide set by scoring the heteroclitic base set (second peptide set) and filtering the obtained scores to compare the peptide-HLA immunogenicity score or peptide-HLA binding score of the peptides for one or more HLA alleles to a threshold. In some embodiments, an affinity criterion of about 50 nM is used to increase the probability that the vaccine peptides will be found and presented by HLA molecules. In some embodiments, the affinity criterion is stricter than 50 nM (i.e., less than 50 nM). In some embodiments, the affinity criterion is stricter than about 500 nM (i.e., less than 500 nM). In some embodiments, as the individual peptide-HLA binding scores or immunogenicity indexes are determined, peptides may be retained as long as they meet the criteria for at least one HLA allele, and only peptide-HLA scores that meet the criteria are considered for vaccine design.

[0686] In some embodiments, the next step is to input the second peptide set into OptiVax to select a compact vaccine peptide set that maximizes predicted vaccine performance (vaccine performance optimization, FIG. 1-2). In some embodiments, the predicted vaccine performance is a function of expected peptide-HLA binding affinity (e.g., a function of the distribution of peptide-HLA binding affinity across all peptide-HLA combinations for a given peptide set, or a function corrected for the occurrence of HLA alleles in a population or individual). In some embodiments, the predicted vaccine performance is the expected population coverage of the vaccine. In some embodiments, the predicted vaccine performance is the expected number of peptide-HLA hits produced by the vaccine in a population or individual. In some embodiments, the predicted vaccine performance requires a minimum expected number of peptide-HLA hits produced by the vaccine (e.g., 1, 2, 3, 4, 5, 6, 7, 8 or more). In some embodiments, the predicted vaccine performance is a function of the population coverage and the number of peptide-HLA hits expected to be produced by the vaccine. In some embodiments, the predicted vaccine performance is a measure of the overall immunogenic properties of the vaccine, where all peptides in the vaccine are scored for peptide-HLA immunogenicity for two or more HLA alleles (e.g., three or more HLA alleles). In some embodiments, the predicted vaccine performance excludes the immunogenic contribution of selected HLA alleles that exceed the maximum number of peptide-HLA hits (e.g., 1, 2, 3, 4, 5, 6, 7, 8 or more). In some embodiments, the predicted vaccine performance excludes the immunogenic contribution of individual HLA diplotypes that exceed the maximum number of peptide-HLA hits (e.g., 1, 2, 3, 4, 5, 6, 7, 8 or more). In some embodiments, the predicted vaccine performance is the proportion of HLA alleles of interest, which is the expected proportion of HLA alleles in each individual that will produce the minimum number of peptides with peptide-HLA immunogenicity expected to be produced by the vaccine (e.g., 1, 2, 3, 4, 5, 6, 7, 8 or more).In some embodiments, the predicted vaccine performance is the expected proportion of HLA alleles in each individual that have a minimum number of peptides with peptide-HLA immunogenicity expected to be produced by the vaccine (e.g., 1, 2, 3, 4, 5, 6, 7, 8 or more).

[0687] In some embodiments, vaccines are designed by iterative selection from a heteroclitic base set (also called an expansion set, as shown in Figures 1-2) with increasingly less stringent criteria for predicted peptide immunogenicity or presentation. In some embodiments, peptides are retained if at least one of their peptide-HLA scores is not eliminated by the adopted threshold. In some embodiments, OptiVax is first used to design vaccines with specific peptide qualification criteria (e.g., seed peptide-HLA scores from the candidate set must bind at least one MHC molecule ≥ 500 nM, and peptide-HLA scores from the expansion set must bind at least one MHC molecule ≥ 50 nM). The vaccines resulting from the application of OptiVax are then used as the basis for vaccine enhancement with less stringent criteria (e.g., seed peptide-HLA scores from the candidate set must bind at least one MHC molecule ≥ 1000 nM, and peptide-HLA scores from the expansion set must bind at least one MHC molecule ≥ 100 nM) to further improve the desired vaccine performance. Methods for vaccine boosting are described in Liu et al. (2020b), which is incorporated by reference in its entirety into this application. In some embodiments, multiple vaccine boosting regimens can be utilized. In some embodiments, the selected vaccine is the final boosted vaccine.

[0688] In some embodiments, the selection of peptide sets that meet the expected performance of the desired vaccine can be performed by a computational algorithm other than OptiVax. In some embodiments, integer linear programming or mixed integer linear programming is used to select peptide sets instead of OptiVax. An example of an integer programming method for peptide set selection is described by Toussaint et al. (2008), which is incorporated by reference in its entirety. An example of a mixed integer linear programming solution is Python-MIP, which can be used in combination with Toussaint et al. (2008). A second example of a method for selecting vaccine peptides is described in "Maximum n-times Coverage for Vaccine Design" by Liu et al. (2021), which is incorporated by reference in its entirety.

[0689] The predicted vaccine performance refers to an index. The predicted vaccine performance can be expressed as a single number, multiple numbers, any number of non-numeric values, and combinations thereof. The value or values ​​can be expressed in any mathematical or symbolic term and in any scale (e.g., nominal scale, ordinal scale, interval scale, ratio scale, etc.).

[0690] A seed (or base) peptide and all modified peptides derived from that seed (or base) peptide constitute a single peptide family. In some embodiments, in a component of vaccine performance based on peptide-HLA immunogenicity for a given HLA allele, a maximum number of peptides (e.g., 1, 2, 3, 4, 5, 6, 7, 8 or more) in the same peptide family are given recognition for their immunogenicity calculated for that HLA allele. This restriction on the immunogenicity of a peptide family limits the recognition of many modified versions of the same base peptide. In some embodiments, the methods described herein include a method for running OptiVax with an EvalVax objective function that corresponds to a desired metric of predicted vaccine performance. In some embodiments, population coverage refers to the proportion of a subject population that presents one or more immunogenic peptides that activate T cells in response to the seed (or base) target peptide. A measure of population coverage is calculated using HLA haplotype frequencies in a particular population, such as a representative human population. In some embodiments, a measure of population coverage is calculated using a threshold HLA frequency in the population. Maximizing population coverage means selecting a peptide set (a base peptide set, a modified peptide set, or a combination of base and modified peptides, e.g., a first peptide set, a second peptide set, or a third peptide set) that represents the greatest proportion of the population with at least a minimum number (e.g., 1, 2, 3, 4, 5, 6, 7, 8, or more) of immunogenic peptide-HLA bindings based on the proportion of HLA haplotypes in a particular population (e.g., a representative human population). In some embodiments, this process includes selecting heteroclitic peptides (as described in this disclosure) with OptiVax that activate T cells that respond to the corresponding seed (or base) peptide and heteroclitic base peptide to improve population coverage. In some embodiments, the seed (or base) target peptide is always included in the final vaccine design.In some embodiments, a peptide is considered a candidate for vaccine design (e.g., included in the first, second, and / or third peptide set) only if it is observed to be immunogenic in clinical data, animal models, or tissue culture models.

[0691] In this disclosure, a heteroclitic peptide is used as an exemplary embodiment, but any modified peptide can be used instead of the heteroclitic peptide. A modified peptide is a peptide that has one or more amino acid substitutions of the target base / seed peptide. The amino acid substitutions can be located at anchor positions or other non-anchor positions.

[0692] In some embodiments, a candidate vaccine peptide (e.g., a basic or modified peptide) is removed from inclusion in the vaccine if it activates T cells that recognize self-peptides (e.g., this can be accomplished in a first and / or second round of peptide filtering and sorting, as shown in Figures 1-2). In some embodiments, a candidate vaccine peptide (e.g., a basic or modified peptide) is computationally removed from inclusion in the vaccine if its outward-facing amino acids when bound by an HLA allele are similar to outward-facing self-peptide residues presented by the same HLA allele, where similarity can be defined by identity or a defined similarity index such as a BLOSUM matrix (BLOSUM matrices are known in the art). Testing of vaccine peptides for their ability to activate T cells that recognize self-peptides can be done experimentally using human tissue protocols, or by vaccination of animal models followed by ELISPOT or other immunogenicity assays. In either case, models are used that have HLA alleles that present the vaccine peptide. In some embodiments, human mononuclear cells (PBMCs) are stimulated with vaccine peptides, T cells are expanded, and activation of T cells by self-peptides is assayed as described in Tapia-Calle et al. (2019) or other methods known in the art. In some embodiments, if T cells are activated by the self-peptide, the vaccine peptide is excluded from inclusion in the vaccine. In some embodiments, computational predictions of the peptide's ability to activate T cells that also recognize the self-peptide can be utilized. These predictions can be made based on modeling the interactions of outward-facing residues of the peptide-HLA complex with other peptide residues. In some embodiments, a candidate vaccine peptide (e.g., a basic or modified peptide) is excluded from vaccine inclusion or experimentally tested for cross-reactivity if it is predicted to activate T cells that also recognize the self-peptide based on the structural similarity of the peptide-MHC complex of the candidate peptide (e.g., a basic or modified peptide) and the peptide-MHC complex of the self-peptide.One method for predicting peptide-MHC structures is that described by Park et al. (2013).

[0693] In some embodiments, if a candidate heteroclitic vaccine peptide does not activate T cells that recognize the corresponding base / seed target peptide of a particular HLA allele (second round of peptide scoring and score filtering, FIG. 1-2), the peptide-HLA binding score or peptide-HLA immunogenicity index of the candidate heteroclitic vaccine peptide (e.g., modified peptide) and HLA allele is excluded from consideration in vaccine design. In some embodiments, if a candidate heteroclitic vaccine peptide does not activate T cells that recognize the corresponding base / seed target peptide (second round of peptide scoring and score filtering, FIG. 1-2) for a given HLA allele, the heteroclitic vaccine peptide (e.g., modified peptide) is excluded from vaccine design. Testing the ability of a candidate heteroclitic peptide (e.g., modified peptide) to activate T cells that recognize the corresponding seed (or base) target peptide for the same HLA allele can be done by vaccination of an animal model followed by ELISPOT or other immunogenicity assays, or experimentally using human tissue protocols. In either case, a model with an HLA allele that presents the heteroclitic peptide is used. In some embodiments, human PBMCs are stimulated with heteroclitic peptides, T cells are expanded, and then activation of T cells by seed (or base) target peptides is assayed as described by Tapia-Calle et al. (2019) or using other methods known in the art. In some embodiments, computational predictions of the ability of heteroclitic peptides to activate T cells that also recognize the corresponding seed (or base) target peptides can be utilized. These predictions can be made based on modeling the interactions of outward-facing residues of peptide-HLA complexes with other peptide residues.In some embodiments, structural similarity between the peptide-HLA complex of a heteroclitic peptide and the peptide-HLA complex of the corresponding seed (or base) target is used to qualify the heteroclitic peptide for vaccine inclusion or to require experimental immunogenicity testing prior to vaccine inclusion.

[0694] TCR binding interface diversity (TCRID) is the minimum mean square deviation of the difference between the 3D positions of the TCR pair residues of a first peptide for a particular HLA allele and the corresponding residue positions of a second peptide. In...

Claims

1. 1. A composition comprising one or more polynucleotides encoding a fragment of a KRAS protein, the fragment of the KRAS protein comprises a G12D mutation and a heteroclitic modification; The composition, wherein the one or more polynucleotides encode at least two amino acid sequences selected from the group consisting of SEQ ID NO:170, SEQ ID NO:173, SEQ ID NO:174, SEQ ID NO:175, SEQ ID NO:176, SEQ ID NO:8439, SEQ ID NO:8465, SEQ ID NO:9059, SEQ ID NO:9067, SEQ ID NO:9106, SEQ ID NO:9642, SEQ ID NO:9698, SEQ ID NO:9721.

2. The composition of claim 1, wherein the one or more polypeptides are included in a construct for expressing at least two peptides encoded by the one or more polynucleotides in vivo in a subject.

3. The composition of claim 2, wherein administration of the one or more polynucleotides enables expression of at least two peptides encoded by the one or more polynucleotides that are presented by HLA class I molecules in a subject.

4. Administration of the one or more polynucleotides a first peptide of said at least two peptides presented by a first plurality of HLA class I alleles; a second peptide of said at least two peptides presented by a second plurality of HLA class I alleles; Enables the expression of The composition of claim 3, wherein the first plurality of HLA class I alleles and the second plurality of HLA class I alleles differ by at least one HLA class I allele.

5. The composition of claim 1, wherein the composition is an immunogenic composition and is for preventing or treating cancer in a subject.

6. 1. A composition comprising one or more polynucleotides encoding a fragment of a KRAS protein, the fragment of the KRAS protein comprises a G12V mutation and a heteroclitic modification; The composition, wherein the one or more polynucleotides encode at least two amino acid sequences selected from the group consisting of the following SEQ ID NO:11018, SEQ ID NO:11054, SEQ ID NO:11078, SEQ ID NO:11080, SEQ ID NO:11447, SEQ ID NO:11468, SEQ ID NO:11469, SEQ ID NO:11474.

7. The composition of claim 6, wherein the one or more polypeptides are included in a construct for expressing at least two peptides encoded by the one or more polynucleotides in vivo in a subject.

8. The composition of claim 7, wherein administration of the one or more polynucleotides enables expression of at least two peptides encoded by the one or more polynucleotides that are presented by HLA class I molecules in a subject.

9. Administration of the one or more polynucleotides a first peptide of said at least two peptides presented by a first plurality of HLA class I alleles; allowing expression of a second peptide of said at least two peptides presented by a second plurality of HLA class I alleles; 9. The composition of claim 8, wherein the first plurality of HLA class I alleles and the second plurality of HLA class I alleles differ by at least one HLA class I allele.

10. The composition of claim 6, wherein the composition is an immunogenic composition and is for preventing or treating cancer in a subject.

11. 1. A composition comprising one or more polynucleotides encoding a fragment of a KRAS protein, the fragment of the KRAS protein comprises a G12R mutation and a heteroclitic modification; The composition, wherein the one or more polynucleotides encode at least two amino acid sequences selected from the group consisting of the following SEQ ID NO:188, SEQ ID NO:189, SEQ ID NO:9760, SEQ ID NO:9772, SEQ ID NO:9775, SEQ ID NO:9779, SEQ ID NO:9785, SEQ ID NO:10076, SEQ ID NO:10103, SEQ ID NO:10227.

12. The composition of claim 11, wherein the one or more polypeptides are included in a construct for expressing at least two peptides encoded by the one or more polynucleotides in vivo in a subject.

13. The composition of claim 12, wherein administration of the one or more polynucleotides enables expression of at least two peptides encoded by the one or more polynucleotides that are presented by HLA class I molecules in a subject.

14. Administration of the one or more polynucleotides a first peptide of said at least two peptides presented by a first plurality of HLA class I alleles; allowing expression of a second peptide of said at least two peptides presented by a second plurality of HLA class I alleles; 14. The composition of claim 13, wherein the first plurality of HLA class I alleles and the second plurality of HLA class I alleles differ by at least one HLA class I allele.

15. The composition of claim 11, wherein the composition is an immunogenic composition and is for preventing or treating cancer in a subject.

16. 1. A composition comprising one or more polynucleotides encoding a fragment of a KRAS protein, the fragment of the KRAS protein comprises a G12C mutation and a heteroclitic modification; The composition, wherein the one or more polynucleotides encode at least two amino acid sequences selected from the group consisting of SEQ ID NO:163, SEQ ID NO:164, SEQ ID NO:7921, SEQ ID NO:7933, SEQ ID NO:7950, SEQ ID NO:7951, SEQ ID NO:8272, SEQ ID NO:8296, SEQ ID NO:8422.

17. The composition of claim 16, wherein the one or more polypeptides are included in a construct for expressing at least two peptides encoded by the one or more polynucleotides in vivo in a subject.

18. The composition of claim 17, wherein administration of the one or more polynucleotides enables expression of at least two peptides encoded by the one or more polynucleotides that are presented by HLA class I molecules in a subject.

19. Administration of the one or more polynucleotides a first peptide of said at least two peptides presented by a first plurality of HLA class I alleles; allowing expression of a second peptide of said at least two peptides presented by a second plurality of HLA class I alleles; 20. The composition of claim 18, wherein the first plurality of HLA class I alleles and the second plurality of HLA class I alleles differ by at least one HLA class I allele.

20. The composition of claim 16, wherein the composition is an immunogenic composition and is for preventing or treating cancer in a subject.