Novel tumor-specific antigens and their use against ovarian cancer

Abstract

This provides a novel tumor-specific antigen (TSA) shared by the majority of ovarian tumors. [Solution] Most (>80%) of the TSAs described herein are derived from abnormally expressed non-mutant genomic sequences, such as intronic sequences and intergenetic sequences, which are not expressed in normal tissues. Nucleic acids, compositions, cells, and vaccines derived from these TSAs are described. The use of TSAs, nucleic acids, compositions, cells, and vaccines for the treatment of ovarian cancer is also described.

Description

[Technical Field]

[0001] Cross-reference of related applications This application is incorporated herein by reference by the U.S. Patent Application filed on June 25, 2019. We assert the rights of the provisional national patent application No. 62 / 866,089.

[0002] This disclosure relates, in general, to cancer, and more specifically, to T cell-based cancer immunity. Regarding tumor antigens specific to ovarian cancer that are useful in epidemic therapy. [Background technology]

[0003] Ovarian cancer is the leading cause of gynecological malignancy deaths worldwide, with 14,000 cases reported annually in the United States. It has caused more than 0,000 deaths (1). High-grade serous ovarian cancer (HGSC) These causes account for 70-80% of deaths, and overall survival has not changed significantly over several decades. (2) A positive correlation was found between the abundance of tumor-infiltrating lymphocytes (TILs) and increased overall survival. The relationship is that T cells can recognize biologically related tumor antigens in HGSCs. This suggests (3, 4). Furthermore, strong evidence is that HGSCs adjacent to tumor epithelial cells This suggests that TILs are actively involved in local immunoediting. In fact, 38 patients In a multimodality study of 212 HGSC samples from CD8 + TIL It was negatively associated with malignant cell diversity (5). Accordingly, and also with several tumors Considering the therapeutic efficacy of immune checkpoint inhibitors in this type, one or more checkpoints Clinical trials using cytotoxic inhibitors are currently underway at HGSC. However, Furthermore, early studies using anti-PD1 have shown that its activity in HGSCs is limited. (6, 7)

[0004] Considering this, it is possible to induce a therapeutic immune response again in ovarian tumors such as HGSCs. It is urgent to identify antigens that can be used in vaccines (3, 8). Such antigens are vaccines. (± immune checkpoint inhibitors), or T cell receptor-based approaches ( It can be used as a target for cytotherapy (bispecific biological agents) (9).

[0005] This explanation refers to several documents, and their contents are based on references to the original source. It will be included in the detailed specifications. [Overview of the project]

[0006] This disclosure provides the following items 1 to 61. 1. A tumor antigen peptide containing one of the amino acid sequences described in SEQ ID NOs: 1-103. 2. The tumor described in item 1, containing one of the amino acid sequences described in SEQ ID NOs: 19-103. Cerebral antigen peptide. 3. The tumor antigen peptide binds to the HLA-A*01:01 molecule, as shown in SEQ ID NOs. 21 and 28. , containing the amino acid sequence described in 40, 41, 66 or 88, the tumor described in item 1 or 2 Cerebral antigen peptide. 4. The tumor antigen peptide binds to the HLA-A*02:01 molecule, and SEQ ID NOs: 1, 19, 20, 22, 30, 31, 36, 50, 52, 60, 62, 73, 84, 85, 86 also The tumor antigen peptide described in item 1 or 2 comprises the amino acid sequence described in 91. 5. The tumor antigen peptide binds to the HLA-A*11:01 molecule, as shown in SEQ ID NOs. 32 and 54. Items including the amino acid sequence described in 55, 67, 69, 81, 87, 90, or 102 The tumor antigen peptide described in 1 or 2. 6. The tumor antigen peptide binds to the HLA-A*24:02 molecule, and SEQ ID NO: 33 or A tumor antigen peptide as described in item 1 or 2, comprising the amino acid sequence described in 43. 7. The tumor antigen peptide binds to the HLA-A*25:01 molecule, as described in Sequence ID No. 24. A tumor antigen peptide as described in item 1 or 2, comprising one of the amino acid sequences of the following. 8. The tumor antigen peptide binds to the HLA-A*29:02 molecule, and SEQ ID NO: 34 or A tumor antigen peptide described in item 1 or 2, containing one of the amino acid sequences described in 58. Do. 9. The tumor antigen peptide binds to the HLA-A*32:01 molecule, as described in Sequence ID No. 16. A tumor antigen peptide as described in item 1 or 2, comprising the amino acid sequence of the following: 10. The tumor antigen peptide binds to the HLA-B*07:02 molecule, and SEQ ID NOs: 4, 6, The item includes the amino acid sequence described in 8, 9, 26, 49, 78, 92, 97, or 101. Tumor antigen peptide as described in item 1 or 2. 11. The tumor antigen peptide binds to the HLA-B*08:01 molecule, SEQ ID NOs: 23, 35 , the amino acids described in 42, 44, 46, 59, 63, 70, 74, 76, 83 or 103 A tumor antigen peptide as described in item 1 or 2, comprising one of the acid sequences. 12. The tumor antigen peptide binds to the HLA-B*14:01 molecule, as indicated in Sequence ID No. 53. A tumor antigen peptide as described in item 1 or 2, containing the amino acid sequence shown. 13. The tumor antigen peptide binds to the HLA-B*15:01 molecule, and SEQ ID NOs: 2, 3 A tumor antigen peptide as described in item 1 or 2, comprising the amino acid sequence described in item 5. 14. The tumor antigen peptide binds to the HLA-B*18:01 molecule, as indicated in Sequence ID No. 89. A tumor antigen peptide as described in item 1 or 2, containing the amino acid sequence shown. 15. The tumor antigen peptide binds to the HLA-B*39:01 molecule, SEQ ID NO: 47, 6 4, 96 or 99, comprising the amino acid sequence described in item 1 or 2, the tumor antigen peptide described in item 1 or 2. Chido. 16. The tumor antigen peptide binds to the HLA-B*40:01 molecule, SEQ ID NO: 11, 1 A tumor antigen peptide as described in item 1 or 2, comprising the amino acid sequence described in item 2 or 13. 17. The tumor antigen peptide binds to the HLA-B*44:02 molecule, as indicated in Sequence ID No. 65. A tumor antigen peptide as described in item 1 or 2, containing the amino acid sequence shown. 18. The tumor antigen peptide binds to the HLA-B*44:03 molecule, and SEQ ID NO: 37 The tumor antigen peptide described in item 1 or 2 comprises the amino acid sequence described in 94. 19. The tumor antigen peptide binds to the HLA-C*03:03 molecule, and SEQ ID NO: 10, 2 The tumor antigen peptide described in item 1 or 2, comprising the amino acid sequence described in 9, 71, or 95. Chido. 20. The tumor antigen peptide binds to the HLA-C*04:01 molecule, and SEQ ID NO: 6 or A tumor antigen peptide as described in item 1 or 2, comprising the amino acid sequence described in item 15. 21. The tumor antigen peptide binds to the HLA-C*05:01 molecule, as indicated in Sequence ID No. 27. A tumor antigen peptide as described in item 1 or 2, containing the amino acid sequence shown. 22. The tumor antigen peptide binds to the HLA-C*06:02 molecule, and SEQ ID NO: 18 The tumor antigen peptide described in item 1 or 2 comprises the amino acid sequence described in item 72. 23. The tumor antigen peptide binds to the HLA-C*07:01 molecule, SEQ ID NO: 38, 6 A tumor antigen peptide as described in item 1 or 2, comprising the amino acid sequence described in 1 or 93. 24. The tumor antigen peptide binds to the HLA-C*07:02 molecule, as described in Sequence ID No. 7. A tumor antigen peptide as described in item 1 or 2, comprising the amino acid sequence of the following: 25. The tumor antigen peptide binds to the HLA-C*12:03 molecule, as indicated in Sequence ID No. 80. A tumor antigen peptide as described in item 1 or 2, containing the amino acid sequence shown. 26. The tumor antigen peptide binds to the HLA-C*14:02 molecule, SEQ ID NO: 25, 5 A tumor antigen peptide as described in item 1 or 2, comprising the amino acid sequence described in 7 or 79. 27. Encoded by a sequence located in the non-protein coding region of the genome, sequence number 1 , 4, 6-8, 10, 13, 15-27 and 36-99, preferably sequence numbers 19-2 Items 1-26 include the amino acid sequence described in 7 and any one of 36-99. The tumor antigen peptide described in any one of the items. 28. The non-protein-coding region of the genome is an untranslated transcription region (UTR), and the tumor The antigen peptide is preferably sequence number 1, 8, 10, 13, 15, and 19-27. The amino acid sequence described in item 27 includes one of the amino acid sequences described in any one of the column numbers 19-27. Tumor antigen peptide. 29. The non-protein-coding region of the genome is an intron, and the tumor antigen peptide is , Sequence IDs 16, 17, and 36-64, preferably any of Sequence IDs 36-64 A tumor antigen peptide as described in item 27, comprising any one of the amino acid sequences described in one of the items. 30. The non-protein-coding region of the genome is an intergenetic region, and the tumor antigen peptide However, item 27 contains the amino acid sequence described in any one of sequence numbers 65-84. The tumor antigen peptide described. 31. The non-protein-coding region of the genome is the source of non-coding RNA transcripts (ncRNAs). The tumor antigen peptide is a xon, and the tumor antigen peptide is sequence numbers 4 and 85-92, preferably sequence numbers 4, 85-92. Tumors as described in item 27, containing the amino acid sequence described in any one of numbers 85-92. Antigen peptide. 32. The non-protein-coding region of the genome is the antisense strand of the gene, and the tumor antibody The proto-peptide contains the amino acid sequence described in any one of SEQ ID NOs: 93-99. The tumor antigen peptide described in item 27. 33. A nucleic acid encoding a tumor antigen peptide as described in any one of items 1 to 32. 34. Nucleic acids, as described in item 33, which are mRNA or viral vectors. 35. A tumor antigen peptide as described in any one of items 1 to 32, or item 33 or Liposomes containing the nucleic acids described in 34. 36. Tumor antigen peptide as described in any one of items 1 to 32, or item 31 or 32 The nucleic acids described, or the liposomes described in item 33, and a pharmaceutically acceptable carrier. , composition. 37. Tumor antigen peptide as described in any one of items 1-32, or item 33 or 34 The nucleic acids described, the liposomes described in item 35, or the compositions described in item 36, and Aji Vaccines, including those containing IVB. 38. The tumor antigen peptide described in any one of items 1 to 32 within the peptide bond groove Includes isolated major histocompatibility complex (MHC) class I molecules. 39. An isolated MHC class I molecule described in item 38, in the form of a multimer. 40. The isolated MHC class I molecule described in item 39, wherein the polymer is a tetramer. 41. (i) A tumor antigen peptide as described in any one of items 1 to 32, or (ii) item Includes a nucleotide sequence encoding a tumor antigen peptide as described in any one of items 1 to 32. Isolated cells containing a vector. 42. The tumor antigen peptides described in any one of items 1 to 32 within the peptide binding grooves A single major histocompatibility complex (MHC) class I molecule containing a single gene is expressed on its surface. Separated cells. 43. Antigen-presenting cells (APCs), as described in item 42. 44. The cell described in item 43, wherein the APC is a dendritic cell. 45. Isolated MHC class I molecules and / or as described in any one of items 38-40. or MHC class I molecules expressed on the surface of cells as described in any one of items 42-44 T cell receptors (TCRs) that specifically recognize [certain cells]. 46. ​​Isolated CD8 cells expressing the TCR described in item 45 on their cell surface. + T lymphocytes. 47. CD8 defined in item 46, at least 0.5% + A group of cells including T lymphocytes . 48. A method for treating ovarian cancer in the subject, wherein an effective amount of (i) items 1 to 32 (ii) Tumor antigen peptide as described in any one of the items, (ii) Nucleic acid as described in item 33 or 34, (iii) Liposomes as described in item 35, (iv) Compositions as described in item 36, (v) (vi) The vaccine described in item 37, (vii) The cells described in any one of items 41 to 45, (vii ) CD8 as described in item 46 + T lymphocytes, or the cell populations described in item (viii) 47. A method comprising administering to a subject. 49. The method according to item 48, wherein the ovarian cancer is serous carcinoma. 50. The method according to item 49, wherein the serous carcinoma is a high-grade serous carcinoma (HGSC). . 51. Further comprising administering at least one additional antitumor agent or therapy to the subject. or the method described in any one of items 48-50. 52. The at least one additional antitumor agent or therapy is a chemotherapeutic agent, immunotherapy, or immunotherapy agent. The method described in item 51, which is checkpoint inhibitor, radiotherapy, or surgery. 53. (i) Tumor antigen peptide as described in any one of items 1 to 32, (ii) Item 33 Alternatively, nucleic acids as described in item 34, (iii) liposomes as described in item 35, (iv) item 36 (v) The composition described in item 37, (vi) Any of items 41-45 (vii) The cells described in item 1, CD8 described in item 46 + T lymphocytes, or (viii ) Use of the cell population described in item 47 for the treatment of ovarian cancer in subjects. 54. (i) Tumor antigen peptide as described in any one of items 1 to 32, (ii) Item 33 Alternatively, nucleic acids as described in item 34, (iii) liposomes as described in item 35, (iv) item 36 (v) The composition described in item 37, (vi) Any of items 41-45 (vii) The cells described in item 1, CD8 described in item 46 + T lymphocytes, or (viii ) The manufacture of pharmaceuticals for the treatment of ovarian cancer in the cell population described in item 47. For use. 55. Use as described in item 53 or 54, wherein the ovarian cancer is serous carcinoma. 56. Use as described in item 55, where the serous carcinoma is a high-grade serous carcinoma (HGSC) . 57. Further including the use of at least one additional antitumor agent or therapy, items 53-56 Use as described in any one of the items. 58. The use according to item 57, wherein the at least one additional anti-tumor agent or therapy is a chemotherapeutic agent, immunotherapy, immune checkpoint inhibitor, radiotherapy or surgery. 59. For use in treating ovarian cancer in a subject, (i) a tumor antigen peptide according to any one of items 1 to 32, (ii) a nucleic acid according to item 33 or 34, (iii) a liposome according to item 35, (iv) a composition according to item 36, (v) a vaccine according to item 37, (vi) a cell according to any one of items 41 to 45, (vii) a CD8 T lymphocyte according to item 46, or (viii) a cell population according to item 47. + 60. A tumor antigen peptide, nucleic acid, liposome, composition, vaccine, cell, CD8 T lymphocyte, or cell population for use according to item 59, wherein the ovarian cancer is serous cancer. + 61. A tumor antigen peptide, nucleic acid, liposome, composition, vaccine, cell, CD8 T lymphocyte, or cell population for use according to item 60, wherein the serous cancer is high-grade serous cancer (HGSC). + 62. A tumor antigen peptide, nucleic acid, liposome, composition, vaccine, cell, CD8 + T lymphocyte, or cell population for use according to any one of items 59 to 61, wherein the tumor antigen peptide, nucleic acid, liposome, composition, vaccine, cell, CD8 T lymphocyte, or cell population is used in combination with at least one additional anti-tumor agent or therapy. + 63. The use according to item 62, wherein the at least one additional anti-tumor agent or therapy is a chemotherapeutic agent, immunotherapy, immune checkpoint inhibitor, radiotherapy or surgery. Cerebral antigen peptide, nucleic acid, liposome, composition, vaccine, cell, CD8 + T lymphocytes, A group of cells.

[0007] Other objects, advantages, and features of the present invention are given only as illustrative examples with reference to the accompanying drawings. This will become apparent by reading the non-restrictive description of the following specific embodiments. [Brief explanation of the drawing]

[0008] [Figure 1] This outlines the workflow of the TSA identification pipeline used in this study. HGSC samples were processed for immunoprecipitation and RNA sequencing. Peptide sequences were identified using MS analysis, which identified MAPs by searching for matches within customized individual global cancer databases constructed from RNA-Seq data. CDS: coding sequence. [Figure 2] Figures 2A and 2B show the expression of RNA encoding candidate aeTSA in normal tissues. These are heatmaps showing the average RNA expression of aeTSA coding sequences in 27 peripheral tissues, with color intensity corresponding to the expression level in each tissue for every 100 million reads (average logarithm-transformed reads (rphm)). Bold boxes indicate tissues / organs where the threshold RNA expression (average rphm > 10) is exceeded. The number next to each peptide sequence indicates the number of tissues where the corresponding RNA expression exceeds the threshold. [Figure 3]Figures 3A–3D show that most TSAs originate from non-mutated non-exon sequences. Figure 3A: Number of MAPs (left) and TSAs (right) identified in each sample. Figure 3B: Scatter plot showing the Pearson correlation between the number of MAPs and TSAs identified per sample. Figure 3C: Bar graph showing the origin of TSAs identified in the cohort studied herein and the Schuster et al. dataset. Blue shades indicate the number of TSAs resulting from in-frame exon translation (coding in), out-frame exon translation (coding out), or non-exon translation (non-coding). Figure 3D: Pie chart showing the translation frame of aeTSA (inner circle), detailed genomic origin (center circle), and their reporting status (outer circle). [Figure 4] This shows the expression of aeTSA coding regions across ovarian cancer samples. The heatmap shows RNA expression of each aeTSA coding region in the nine samples reported in this study (left) and 378 samples from the TCGA-OV cohort, and the color intensity indicates the RNA level in the mapped reads per million reads. [Figure 5] Figures 5A and 5B show that changes in copy number correlate with the expression of several aeTSA. Figure 5A: The heatmap shows the Spearman correlation between aeTSA RNA expression levels and DNA copy number, promoter methylation, or gene expression for aeTSA located intragenetically (left) or extrageneically (right). Unavailable data are shown in light gray. Figure 5B: Number of aeTSA identified from each chromosome arm (top) with arm-level amplification scores (bottom). Asterisks indicate amplification considered significant (Q value < 0.25). [Figure 6]Figures 6A–6E show that presentation of three aeTSAs can induce a spontaneous antitumor immune response. Figures 6A–6C: Kaplan-Meier curves show the survival of four groups of patients in the TCGA-OV cohort. For each of the three individual aeTSAs, one group was able to present aeTSA(EP) and three groups were not (ED, ND, and NP). ED: Expression of aeTSA coding RNA in the absence of the relevant HLA allotype; ND: No expression of aeTSA coding RNA in the absence of the relevant HLA allotype; EP: Expression of aeTSA coding RNA in the presence of the relevant HLA allotype; NP: No expression of aeTSA coding RNA in the presence of the relevant HLA allotype. Color shades represent 95% confidence intervals. Log-rank P values ​​are shown. Figures 6D, 6E: Abundance of T and cytotoxic cells in tumors from the four groups shown in Figure 6C. [Figure 7] This shows the estimated frequencies of aeTSA presented by individual HGSCs in three different populations. One million simulated patients were generated for each population. aeTSA was considered present in simulated patients if its RNA was expressed and the associated HLA allotype was present. The frequency of aeTSA RNA expression was based on TCGA-OV RNA-Seq data (shown in Figure 4). HLA allotype frequencies were obtained from the USA National Marrow Donor Program. The red dashed line shows the median number of aeTSA per tumor in each population. [Figure 8] Figures 8A and 8B demonstrate that mTSA validation based on dbSNP is also supported by sequencing data from paired normal samples. Figure 8A: Example of excluded TSA candidates matching common polymorphisms reported in dbSNP. The mutant nucleotide was also found in paired normal samples. Figure 8B: Example of an mTSA without mutants in dbSNP. The mutant nucleotide is detected only in paired normal samples with one read that is likely a sequencing error. [Figure 9]This shows the peripheral expression of coding sequences for TSA candidates containing germline polymorphisms. TSA candidates with single nucleotide mutations recorded in dbSNPs were considered aeTSA if both their coding sequence and the corresponding reference sequence restricted RNA expression in peripheral tissues. The heatmap shows the mean RNA expression of aeTSA coding sequences in 27 peripheral tissues, where color intensity corresponds to the expression level in each tissue in the mean logarithm-transformed reads (rphm) per 100 million reads. Bold indicates tissues / organs with a mean rphm value greater than 10. The number next to each peptide sequence indicates the number of tissues with significant RNA expression for a given peptide. Red asterisks indicate peptides retained as aeTSA. [Figure 10] This graph shows the correlation between the number of MAPs and tumor size. The scatter plot shows the Pearson correlation between tumor size (x-axis) and MAP number, identified for each HLA allele (y-axis) in each sample. In this plot, only samples from Schuster et al. (the largest subgroup) were used to avoid batch effects. [Figure 11] Figures 11A and 11B are graphs showing the relationship between aeTSA expression and DNA copy number variation (CNV). Figure 11A: Enrichment analysis of significant correlations between intragenetic aeTSA RNA expression and CNV, calculated using Fisher's exact test. aeTSA is grouped based on the proportion of tumors expressing TSA (upper and lower halves). Figure 11B: Correlation between aeTSA number and chromosome arm amplification. The scatter plot shows the Pearson correlation between aeTSA number and chromosome arm amplification (x axis) identified from the arms (y axis) of each sample. [Modes for carrying out the invention]

[0009] The terms and symbols used herein for genetics, molecular biology, biochemistry, and nucleic acids are as follows: , standard papers and texts in the field, e.g., Kornberg and Baker,DNA Replication,Second Edition(WH Freeman, New York, 1992), Lehninger, Bioche mistry,Second Edition(Worth Publishers,N ew York, 1975), Strachan and Read, Human Mo. lecular Genetics,Second Edition(Wiley-Li ss, New York, 1999), Eckstein, editor, Oligon. ucleotides and analogs: A Practical Appro ach(Oxford University Press, New York, 199 1), Gait, editor, Oligonucleotide Synthesis :A Practical Approach(IRL Press,Oxford,1 984) and so on. All terms are those typical of those established in the relevant technical field. It must be understood using that meaning.

[0010] The articles "a" and "an" refer to one or more grammatical objects of the article (i.e., a few Used herein to refer to at least one element. For example, "an elem "ent)" means one or more elements. Throughout this specification, sentence Unless there is a specific reason otherwise, "include (comprise)" The terms "s)" and "comprising" refer to the specified process or This means the inclusion of an element, or a process or group of elements, but any other process or element It will be understood that this does not mean the exclusion of a group of processes or elements. .

[0011] The enumeration of value ranges in this specification means that, unless otherwise indicated herein, the range is within that range. It is intended to serve as a complete and concise way of representing each distinct value that fits within it. Each individual value is merely listed separately herein and is not incorporated herein as if it were. It is included. Also, all subsets of values ​​within the range are as listed individually in this specification. Uni is incorporated herein.

[0012] All methods described herein, unless otherwise indicated herein, or in the context of They can be performed in any preferred order, as long as they do not clearly contradict each other.

[0013] Any and all examples or illustrative language provided herein (e.g., "etc.") The use of ) is intended solely to better illustrate the present invention, unless otherwise claimed. This does not limit the scope of the present invention.

[0014] Any language used herein does not imply any element not claimed to be essential to the implementation of the invention. It should not be interpreted as something that indicates something.

[0015] In this specification, the term "approximately" has its ordinary meaning. The term "approximately" refers to a value. However, this includes inherent variations in error for the device or method used to determine the value. Used to indicate or close to the enumerated values, for example, the enumerated values ​​( It includes values ​​that are within 10% or 5% of the range of values.

[0016] In the research described herein, the inventors used a proteogenome-based approach. Using this method, 111 TSA candidates (103 novel candidates) were selected from 23 HGSC tumors. We identified eight previously reported candidates. The majority of these TSAs (93) were normal. These abnormally expressed non-mutant genome sequences are not expressed in normal tissues. Current TSA (referred to as aeTSA in this specification) mainly consists of non-exon sequences, particularly in It has been shown that these originate from tron ​​(31%) and intergenetic (22%) sequences, and their expression This was regulated at the transcriptional level by variations in gene copy number and DNA methylation. et al. aeTSA is shared by the majority of HGSCs, and the frequency of aeTSA expression and H Considering the LA allele frequency, the median number of aeTSAs per tumor in Caucasians It was estimated to be 5. The novel TSA candidates identified herein are ovarian cancer T cells. This may be useful in immunotherapy based on [the following criteria].

[0017] Therefore, in certain embodiments, this disclosure includes SEQ ID NOs: 1-103, preferably SEQ ID NOs: 19-103. Tumors containing or consisting of one of the amino acid sequences in 03 (Table 3A, Table 3B) This concerns antigen peptides (or tumor-specific peptides).

[0018] In embodiments, this disclosure relates to Sequence IDs 1, 8, 10, 13, 15, and 19-27, Mashiku contains or consists of one of the amino acid sequences of sequence numbers 19-27. , located in the untranslated transcription region (UTR), i.e., the 3'-UTR or 5'-UTR region. This relates to tumor antigen peptides encoded by sequences. In embodiments, tumor antigen peptides This refers to sequence numbers 1, 10, 13, 15, and 19-23, preferably sequence numbers 19-23. A molecule located in the 5'-UTR region that contains or consists of one of the amino acid sequences of the following: It is encoded by the sequence. In this embodiment, the tumor antigen peptide is sequence number 8, and It contains one of the amino acid sequences 24-27, preferably sequence numbers 24-27, or This is encoded by an array located in the 3'-UTR region.

[0019] In embodiments, this disclosure relates to sequence numbers 16, 17, and 36-64, preferably sequence numbers 16, 17, and 36-64. An intron containing or consisting of one of the amino acid sequences number 36-64 This relates to tumor antigen peptides encoded by their positional sequence.

[0020] In embodiments, the present disclosure includes one of the amino acid sequences of SEQ ID NOs. 65 to 84, or tumor antigen peptides, which are encoded by sequences located in intergenetic regions. Regarding Do.

[0021] In another embodiment, the present disclosure relates to Sequence IDs 6, 7, 18 and 28-35, preferably to An exon containing or consisting of one of the amino acid sequences in sequence numbers 28-35. The tumor antigen peptide is located at and encoded by a sequence that begins with a frameshift. To relate to.

[0022] In another embodiment, the present disclosure relates to Sequence ID No. 4 and 85-92, preferably Sequence ID No. 8. Non-coding RNA sequences containing or consisting of one of 5 to 92 amino acid sequences. Tumor antibodies encoded by ncRNAs (located in the exons of non-coding transcripts) Regarding the proto-peptide.

[0023] In another embodiment, the disclosure includes one of the amino acid sequences of SEQ ID NOs: 93-99. or a tumor antisense encoded by a sequence that is an antisense of a gene. Regarding the proto-peptide.

[0024] In another embodiment, the present disclosure uses one of the amino acid sequences of SEQ ID NOs: 100-103 Tumor antigens encoded by sequences from mucin genes, including or consisting of such sequences Regarding Petit Do.

[0025] Generally, peptides such as tumor antigen peptides presented in relation to HLA class I are long The amount is approximately 7 or 8 to approximately 15 amino acid residues, preferably 8 to 14. In some embodiments of the method shown, the tumor antigen peptide sequence is included as defined herein. Longer peptides are artificially loaded into cells such as antigen-presenting cells (APCs), and the cells... The tumor antigen peptide is then processed by MHC class I molecules on the surface of the APC. This is presented. In this method, 15 amino acid residues (i.e., tumor antigen precursor peptide) are presented. Peptides / polypeptides longer than ) can be loaded into the APC, allowing for presentation of the original The proteases in the cytoplasm of APCs that provide the corresponding tumor antigen peptide as defined in the document The tumor antigen peptide as defined herein is processed accordingly. In some embodiments, the tumor antigen peptide as defined herein is used. The precursor peptide / polypeptide used to produce it is, for example, 1000, 500 , 400, 300, 200, 150, 100, 75, 50, 45, 40, 35, 30, 2 The tumor antigen described herein consists of 5, 20, or 15 or fewer amino acids. All methods and processes using peptides are "most" after processing with cells (APCs). Longer peptides are used to induce the presentation of 8-14 tumor antigen peptides, or These are polypeptides (including natural proteins), i.e., tumor antigen precursor peptides / polypeptides. This includes the use of butyl. In some embodiments, the tumor antigen peptide described herein is Approximately 8-14, 8-13, or 8-12 amino acid lengths (for example, 8, 9, 10, 11, (12 or 13 amino acid lengths) and sufficient to directly fit into HLA class I molecules. Small. In the embodiment, the tumor antigen peptide has 20 or fewer amino acids, preferably 15. The following amino acids, more preferably 14 or fewer amino acids, are included. In the embodiment, tumor antigen The peptide consists of at least 7 amino acids, preferably at least 8 amino acids, more preferably It contains at least 9 amino acids.

[0026] As used herein, the term "amino acid" refers to naturally occurring L-amino acids. To prepare both the D-isomer and the D-isomer, as well as synthetic analogs of tumor antigen peptides. Other amino acids used in peptide chemistry (for example, naturally occurring amino acids, naturally occurring It contains amino acids that are not encoded by nucleic acid sequences, such as amino acids that are not encoded by nucleic acid sequences. Examples of amino acids present include glycine, alanine, valine, leucine, isoleucine, and ce Examples include phosphorus and threonine. Other amino acids include, for example, non-genetic forms of amino acids. Examples include the substitution of amino acids and L-amino acids. Naturally occurring non-genetic compounds. Examples of diamino acids include β-alanine, 3-aminopropionic acid, and 2,3-diamino acid. Nopropionic acid, α-aminoisobutyric acid (Aib), 4-amino-butyric acid, N-methylglyceride (Sarcosine), hydroxyproline, ornithine (e.g., L-ornithine), cyto Lurin, t-butylalanine, t-butylglycine, N-methylisoleucine, phenyl Glycine, cyclohexylalanine, norleucine (Nle), norvaline, 2-naphthyl Lalanine, pyridylalanine, 3-benzothienylalanine, 4-chlorophenylalanine Nin, 2-fluorophenylalanine, 3-fluorophenylalanine, 4-fluoro Penylalanine, penicillamine, 1,2,3,4-tetrahydro-isoquinoline-3- Rubonic acid, β-2-thienylalanine, methionine sulfoxide, L-homoarginine ( Hoarg), N-acetyllysine, 2-aminobutyric acid, 2-aminobutyric acid, 2,4-diamycin Nobutyric acid (D- or L-), p-aminophenylalanine, N-methylvaline, homocyan Stain, homoserine (HoSer), cysteic acid, ε-aminohexanoic acid, δ-aminohexanoic acid Examples include novaleric acid or 2,3-diaminobutyric acid (D- or L-). The amino acids are well known in the field of biochemistry / peptide chemistry. In the embodiment, tumor anti The proto-peptide contains only naturally occurring amino acids.

[0027] In embodiments, the tumor antigen peptide described herein is compared with the sequence described herein. The peptide contains a modified sequence that includes functionally equivalent amino acid residue substitutions. For example, one or more amino acid residues in a sequence act as functional equivalents, and siren Alteration occurs when similar polarity (similar physicochemical properties) are present. It can be substituted by another amino acid. Amino acid substitutions in a sequence are those to which the amino acid belongs. Other members of the class may be selected. For example, positively charged (basic) amino acids and For example, arginine, lysine, and histidine (as well as homoarginine and ornithine) Examples include tin. Nonpolar (hydrophobic) amino acids include leucine, isoleucine, and Examples include ranine, phenylalanine, valine, proline, tryptophan, and methionine. It is possible. Examples of uncharged polar amino acids include serine, threonine, cysteine, and tyrosine. Examples include syn, asparagine, and glutamine. These are negatively charged (acidic) amino acids. Examples include glutamic acid and aspartic acid. The amino acid glycine is nonpolar. It may belong to either the amino acid family or the uncharged (neutral) polar amino acid family. Substitutions within an amino acid family are generally understood to be conservative substitutions. The tumor antigen peptides described herein include all L-amino acids and all D-amino acids. , or may include a mixture of L-amino acids and D-amino acids. In embodiments, this specification The tumor antigen peptides described in the book contain all L-amino acids.

[0028] In the embodiment, the sequence includes or is derived from one of the sequences disclosed in Tables 3A to 3B. In the sequence of the tumor antigen peptide, it does not substantially contribute to the interaction with the T cell receptor. The amino acid residues, when incorporated, substantially do not affect T cell reactivity, and are associated with MHC It can be modified by substitution with other amino acids that do not exclude binding to it.

[0029] Tumor antigen peptides also prevent degradation and increase stability, affinity, and / or uptake. To achieve this, the N-terminus and / or C-terminus may be capped or modified. It may be done. Therefore, in another embodiment, this disclosure is formula Z 1 -XZ 2 Modified tumor The formula provides an antigen peptide, where X is SEQ ID NOs: 1-103, preferably SEQ ID NOs: 19-103. A tumor antigen peptide containing or consisting of one of the amino acid sequences of 03. (Table 3A, Table 3B).

[0030] In the embodiment, the amino-terminal residue of the tumor antigen peptide (i.e., the free amino group at the N-terminus) ) is, for example, a partial / chemical group (Z 1 ) through covalent bonds (for example, protection from decomposition) (to be modified). Z 1 This is a linear or branched alkyl group of 1 to 8 carbon atoms, or It may also be a syl group (R-CO-), where R is a hydrophobic moiety (e.g., acetyl, p). (Lopionyl, butanil, isopropionyl, or iso-butanil), or aroyl The group is (Ar-CO-), where Ar is an aryl group. In embodiments, an acyl group C1~C 16 Or C3~C 16 Acyl group (straight-chain or branched, saturated or unsaturated) In further embodiments, saturated C1-C6 acyl groups (linear or branched) or unsaturated C1-C6 acyl groups may be used. A C3-C6 acyl group (linear or branched), for example, an acetyl group (CH3-CO-Ac) Yes. In one embodiment, Z 1 It does not exist. The carboxyl terminal residue of the tumor antigen peptide (sun For example, the free carboxyl group at the C-terminus of the tumor antigen peptide can be amidated (to an NH2 group). It can be modified by substitution of the OH group (for example, to protect from decomposition), so In cases like this, Z 2 This is an NH2 group. In the embodiment, Z 2 is a hydroxamate group, ni Tolyl group, amide (primary, secondary, or tertiary) group, methylamine, isobutylamine, i Aliphatic amines with 1 to 10 carbon atoms, such as sovalerylamine or cyclohexylamine. , aniline, naphthylamine, benzylamine, cinnamylamine, or phenylethylamine Aromatic or arylalkylamines such as armines, alcohols, or CH2OH It may also be Z. In this embodiment, 2 It does not exist. In the embodiment, the tumor antigen peptide is , the amino acid sequences of SEQ ID NOs. 1 to 103, preferably SEQ ID NOs. 19 to 103 (Table 3A, Table 3 Includes one of B). In the embodiment, the tumor antigen peptide is SEQ ID NOs: 1-103, Mashiku consists of one of the amino acid sequences of sequence numbers 19-103 (Table 3A, Table 3B). , in other words, Z 1 and Z 2 It does not exist.

[0031] In another aspect, the present disclosure includes sequences 21, 28, 40, 41, 66, or 88. HLA-A * 01:01 Binds to the molecule, tumor antigen peptide The present invention provides a peptide (or tumor-specific peptide), preferably an ovarian tumor antigen peptide.

[0032] In another aspect, this disclosure relates to Sequence IDs 14, 17, 45, 48, 51, 56, 75, and 77. , 82, 98 or 100, preferably 45, 48, 51, 56, 75, 77, 82, 9 HLA-A, which contains or consists of a sequence of 8 or 100 * 02:01 bonded to the molecule A tumor antigen peptide (or tumor-specific peptide), preferably an ovarian tumor antigen peptide, is combined with the tumor antigen peptide (or tumor-specific peptide). We will provide Chido.

[0033] In another aspect, this disclosure relates to Sequence IDs 1, 19, 20, 22, 30, 31, 36, 50, 52, 60, 62, 73, 84, 85, 86 or 91, preferably 19, 20, 22, Arrays of 30, 31, 36, 50, 52, 60, 62, 73, 84, 85, 86, or 91 HLA-A * 03:01 Tumor antigen peptides that bind to molecules The present invention provides cydo (or tumor-specific peptide), preferably ovarian tumor antigen peptide.

[0034] In another aspect, this disclosure relates to Sequence IDs 32, 54, 55, 67, 69, 81, 87, and 90. Or an HLA-A sequence containing or consisting of 102 * 11:01 Binds to the molecule This involves a tumor antigen peptide (or tumor-specific peptide), preferably an ovarian tumor antigen peptide. We will provide the service.

[0035] In another embodiment, the present disclosure includes or is derived from the sequence of sequence number 33 or 43. HLA-A * 24:02 molecules that bind to tumor antigen peptides (or tumor-specific peptides) The present invention provides a cytoplasmic (Cytoplasmic) peptide, preferably an ovarian tumor antigen peptide.

[0036] In another embodiment, the disclosure includes or comprises the sequence of Sequence ID No. 24, HLA -A * 25:01 A tumor antigen peptide (or tumor-specific peptide) that binds to the molecule, Mashiku provides ovarian tumor antigen peptides.

[0037] In another aspect, the present disclosure includes or is derived from the sequence of sequence number 34 or 58. HLA-A * 29:02 Tumor antigen peptides (or tumor-specific peptides) that bind to molecules. The present invention provides a cytoplasmic (Cytoplasmic) peptide, preferably an ovarian tumor antigen peptide.

[0038] In another aspect, the disclosure includes or comprises the sequence of Sequence ID No. 16, HLA -A * 32:01 A tumor antigen peptide (or tumor-specific peptide) that binds to the molecule, Mashiku provides ovarian tumor antigen peptides.

[0039] In another aspect, this disclosure relates to Sequence IDs 4, 6, 8, 9, 26, 49, 78, 92, and 97. Or it may include an array of 101, preferably 26, 49, 78, 92, 97 or 101, HLA-B * 07:02 Tumor antigen peptides that bind to molecules (or The present invention provides tumor-specific peptides, preferably ovarian tumor antigen peptides.

[0040] In another aspect, this disclosure relates to Sequence IDs 23, 35, 42, 44, 46, 59, 63, and 70. HLA-B * 0 A tumor antigen peptide (or tumor-specific peptide), preferably bound to the 8:01 molecule. This provides ovarian tumor antigen peptides.

[0041] In another aspect, the disclosure includes or comprises the sequence of Sequence ID No. 53, HLA -B * 14:01 A tumor antigen peptide (or tumor-specific peptide) that binds to the molecule, Mashiku provides ovarian tumor antigen peptides.

[0042] In another aspect, the present disclosure includes or is derived from the sequence of sequence numbers 2, 3, or 5. HLA-B * 15:01 Tumor antigen peptides (or tumor-specific peptides) that bind to molecules. The present invention provides a cytoplasmic (Cytoplasmic) peptide, preferably an ovarian tumor antigen peptide.

[0043] In another aspect, the disclosure includes or comprises the sequence of Sequence ID No. 89, HLA -B * 18:01 A tumor antigen peptide (or tumor-specific peptide) that binds to the molecule, Mashiku provides ovarian tumor antigen peptides.

[0044] In another aspect, the present disclosure includes the sequence of sequence numbers 47, 64, 96, or 99, or It consists of HLA-B * 39:01 Tumor antigen peptide (or tumor) that binds to the molecule. The present invention provides a tumor-specific peptide, preferably an ovarian tumor antigen peptide.

[0045] In another aspect, the disclosure includes the sequence of sequence numbers 11, 12, or 13, or the sequence of sequence numbers 11, 12, or 13. HLA-B * 40:01 Tumor antigen peptide (or tumor-specific) that binds to the molecule The present invention provides a peptide, preferably an ovarian tumor antigen peptide.

[0046] In another aspect, the present disclosure includes or comprises the sequence of Sequence ID No. 65, HLA -B * 44:02 A tumor antigen peptide (or tumor-specific peptide) that binds to the molecule, Mashiku provides ovarian tumor antigen peptides.

[0047] In another aspect, the present disclosure includes or is derived from the sequence of sequence number 37 or 94. HLA-B * 44:03 Tumor antigen peptides (or tumor-specific peptides) that bind to molecules. The present invention provides a cytoplasmic (Cytoplasmic) peptide, preferably an ovarian tumor antigen peptide.

[0048] In another aspect, the present disclosure relates to Sequence IDs 10, 29, 71, or 95, preferably Sequence IDs. HLA-C containing or consisting of sequences of 29, 71, or 95 *03:03 minutes A tumor antigen peptide (or tumor-specific peptide) that binds to the tumor, preferably an ovarian tumor antibody We provide the original peptide.

[0049] In another aspect, the disclosure includes or consists of the sequence of sequence number 6 or 15. HLA-C * 04:01 Tumor antigen peptides (or tumor-specific peptides) that bind to molecules. (D) Preferably, the ovarian tumor antigen peptide is provided.

[0050] In another embodiment, the present disclosure includes or comprises the sequence of Sequence ID No. 27, HLA -C * 05:01 A tumor antigen peptide (or tumor-specific peptide) that binds to the molecule, Mashiku provides ovarian tumor antigen peptides.

[0051] In another aspect, the present disclosure relates to Sequence ID No. 18 or 72, preferably Sequence ID No. 18 or 72. HLA-C, which contains or consists of a sequence * 06:02 Tumor antigens that bind to molecules The present invention provides peptides (or tumor-specific peptides), preferably ovarian tumor antigen peptides.

[0052] In another aspect, the present disclosure includes the sequence of sequence numbers 38, 61, or 93, or the sequence of sequence numbers 38, 61, or 93. HLA-C * 07:01 Tumor antigen peptides (or tumor-specific peptides) that bind to molecules The present invention provides a peptide, preferably an ovarian tumor antigen peptide.

[0053] In another aspect, the present disclosure includes or comprises the sequence of Sequence ID No. 7, HLA- C * 07:02 A tumor antigen peptide (or tumor-specific peptide) that binds to the molecule, preferably Alternatively, it provides ovarian tumor antigen peptides.

[0054] In another aspect, the disclosure includes or comprises the sequence of Sequence ID No. 80, HLA -C * 12:03 A tumor antigen peptide (or tumor-specific peptide) that binds to the molecule, Mashiku provides ovarian tumor antigen peptides.

[0055] In another aspect, the present disclosure includes the sequence of sequence numbers 25, 57, or 79, or the sequence of sequence numbers 25, 57, or 79. HLA-C * 14:02 Tumor antigen peptides (or tumor-specific peptides) that bind to molecules The present invention provides a peptide, preferably an ovarian tumor antigen peptide.

[0056] In this embodiment, the tumor antigen peptide is an untranslated transcription region (UTR), i.e., 3'-U It is encoded by a sequence located in the TR or 5'-UTR region. In another embodiment, The tumor antigen peptide is encoded by a sequence located in an intron. In another embodiment, In other words, tumor antigen peptides are encoded by sequences located in intergenetic regions. Morphologically, tumor antigen peptides are located within exons and are derived from frameshifts. Coded by column.

[0057] The tumor antigen peptides of this disclosure include nucleic acids that encode tumor antigen peptides in host cells. It can be produced by expression (recombinant expression) or by chemical synthesis (e.g., solid-phase peptide synthesis). Peptides can be produced using manual and / or automated solid-phase procedures well known in the art. It can be easily synthesized by, for example, "T-boc" or " This can be done by using the "Fmoc" procedure. Techniques for solid-phase synthesis and The procedure is, for example, Solid Phase Peptide Synthesis: A Practical Approach(E.Atherton and RCSh Eppard, author, IRL, Oxford University Press, 19 It is described in (published in 1989). Alternatively, tumor antigen peptides are, for example, Liu e t al., Tetrahedron Lett. 37:933-936, 1996, B aca et al.,J.Am.Chem.Soc.117:1881-1887,1 995, Tam et al., Int. J. Peptide Protein Res .45:209-216,1995, Schnolzer and Kent,Scie. nce 256:221-225, 1992, Liu and Tam, J.Am.Ch em.Soc.116:4149-4153, 1994, Liu and Tam, Pr. oc.Natl.Acad.Sci.USA 91:6584-6588,1994, Yamashiro and Li, Int.J. Peptide Protein As described in Res.31:322-334,1988), segment condensation It may be prepared in this way. Other methods useful for the synthesis of tumor antigen peptides include Nakagawa et al., J.Am.Chem.Soc.107:7087-7092,1985 As described in [reference], in embodiments, the tumor antigen peptide is chemically synthesized (synthetic peptide (d). Another embodiment of the present disclosure is a non-naturally occurring peptide, wherein the peptide is the present Consists of, or essentially comprises, an amino acid sequence as defined in the specification, and is pharmaceutically acceptable. This disclosure relates to synthetically produced (e.g., synthesized) peptides as salts. The salt of the ulcer antigen peptide is not a salt because the peptide produced in vivo is not a salt. This is significantly different from peptides in their natural state. Non-natural salt forms of peptides are particularly relevant in pharmaceuticals containing peptides. In relation to compositions, for example, peptide vaccines disclosed herein, the solubility of the peptide This can be adjusted. Preferably, the salt is a pharmaceutically acceptable salt of the peptide.

[0058] In the embodiments, the tumor antigen peptides described herein are substantially pure. When separated from its naturally associated components, it is "substantially pure." Typically The compound should make up at least 60% by weight, more commonly 75% by weight, and 80% by weight of the total material in the sample. In cases where the amount is % or 85% by weight, preferably more than 90% by weight, more preferably more than 95% by weight Therefore, it is essentially pure. For this reason, for example, it can be chemically synthesized by recombinant technology. Alternatively, the polypeptides produced are generally derived from their naturally occurring components, for example, their supply It will likely contain virtually no components of the macromolecule from which it originates. Nucleic acid molecules are derived from organisms. In naturally occurring genomes, coding sequences that are normally continuous and those that are not immediately contiguous ( In other words, if there are no covalent bonds, it is substantially pure. A substantially pure compound is, for example, For example, extraction from natural sources can lead to the expression of recombinant nucleic acid molecules encoding peptide compounds. Therefore, it can be obtained by chemical synthesis. Purity can be determined by column chromatography. Measurement can be performed using any suitable method such as gel electrophoresis or HPLC. In its morphology, the tumor antigen peptide is in solution. In another embodiment, the tumor antigen peptide is It is in solid form, for example, freeze-dried.

[0059] In another aspect, this disclosure relates to tumor antigen peptides or tumor antigen precursors described herein. Further providing nucleic acids (isolated) encoding ptide. In embodiments, the nucleic acid is approximately 2 1 nucleotide to approximately 45 nucleotides, approximately 24 to approximately 45 nucleotides, for example, 24, 2 Contains 7, 30, 33, 36, 39, 42, or 45 nucleotides. “Isolated” means When used herein, the molecule may contain other components or naturally occurring supplies present in the natural environment. Peptide isolated from source polymers (e.g., other nucleic acids, proteins, lipids, sugars, etc.) Refers to nucleic acid molecules. "Synthetic," as used herein, means, for example, recombinant technology. Peptides produced through art or using chemical synthesis, which are not isolated from their natural sources. This refers to a nucleic acid molecule. The nucleic acids of this disclosure are used for the recombinant expression of tumor antigen peptides of this disclosure. A cloning vector or embryo that can be used to transfect host cells. It may be contained in a vector or plasmid, such as a current vector. In embodiments, this disclosure is, Cloning, expression, or other methods including nucleic acid sequences encoding tumor antigen peptides in this disclosure We provide a tumor vector or plasmid. Alternatively, the tumor antigen peptide of this disclosure can be used as a plasmid. The nucleic acids that are incorporated can be integrated into the genome of the host cell. In either case, the host cell The cells express proteins encoded by tumor antigen peptides or nucleic acids. The term "host cell" as used in this book refers not only to a specific target cell, but also to... This refers to the offspring or potential offspring of a cell. The host cell is the tumor antigen peptide described herein. Any prokaryotic cell (e.g., E. coli) or eukaryotic cell (e.g., For example, it can be an insect cell, yeast cell, or mammalian cell. Rasmid contains elements necessary for the transcription and translation of the inserted coding sequence, and resistance genes It may contain other components such as cloning sites. Using methods well known to those skilled in the art, A sequence encoding a peptide or polypeptide, and an appropriate operably linked thereto Expression vectors containing transcriptional and translational regulatory / modulatory elements may be constructed. These include in vitro recombinant DNA technology, synthesis technology, and in vivo genetic engineering. Such technology is described in Sambrook et al. (1989) Molecul. ar Cloning,A Laboratory Manual,Cold Spri ng Harbor Press, Plainview, NY, and Ausube l,FMet al.(1989)Current Protocols in M Olecular Biology,John Wiley & Sons,New Y It is stated in ork, NY. "Operationally linked" means the components, especially nu This refers to the parallel arrangement of components that enable the creotide sequence to function normally. Therefore, the code sequence that is operably linked to the regulating sequence controls the regulating sequence, that is, Furthermore, nucleotides that can express coding sequences under transcriptional and / or translational control. This refers to the configuration of the control array. The terms "regulatory / control region" or "regulatory / control array" are used herein. In this case, it refers to non-coding nucleotide sequences involved in regulating the expression of coding nucleic acids. The term "regulatory region" refers to a promoter sequence, regulatory protein binding site, and upstream activation. This includes factor sequences, etc. In embodiments, nucleic acids (DN) encoding the tumor antigen peptide of this disclosure. A, RNA) is contained within liposomes or any other suitable vehicle, or It is connected to it.

[0060] In another aspect, the present disclosure includes (i.e., presents or binds to) tumor antigen peptides. (to provide) MHC class I molecules. In the embodiment, the MHC class I molecule is HLA- It is the A1 molecule, and in further embodiments, the HLA-A*01:01 molecule. Morphologically, the MHC class I molecule is an HLA-A2 molecule, and in further embodiments, H It is an LA-A*02:01 molecule. In another embodiment, the MHC class I molecule is HLA- It is the A3 molecule, and in further embodiments, the HLA-A*03:01 molecule. Morphologically, the MHC class I molecule is an HLA-A11 molecule, and in further embodiments, In another embodiment, the MHC class I molecule is HLA -A24 molecule, and in further embodiments, HLA-A*24:02 molecule. In one embodiment, the MHC class I molecule is the HLA-A25 molecule, and in a further embodiment, This is an HLA-A*25:01 molecule. In another embodiment, the MHC class I molecule is H The molecule is LA-A29, and in further embodiments, it is HLA-A*29:02. In another embodiment, the MHC class I molecule is the HLA-A32 molecule, and further embodiments In this embodiment, it is an HLA-A*32:02 molecule. In another embodiment, the MHC class I molecule is In a further embodiment, it is an HLA-B07 molecule, and in a more recent embodiment, an HLA-B*07:02 molecule. In another embodiment, the MHC class I molecule is the HLA-B08 molecule, and further... In the application form, it is an HLA-B*08:01 molecule. In another embodiment, it is an MHC class I molecule. The child is the HLA-B14 molecule, and in further embodiments, the HLA-B*14:01 molecule. In another embodiment, the MHC class I molecule is an HLA-B15 molecule, and further In one embodiment, it is an HLA-B*15:01 molecule. In another embodiment, it is an MHC class Molecule I is the HLA-B18 molecule, and in further embodiments, HLA-B*18:01 It is a molecule. In another embodiment, the MHC class I molecule is the HLA-B39 molecule, In one embodiment, it is the HLA-B*39:01 molecule. In another embodiment, it is the MHC molecule. The LA-I molecule is the HLA-B40 molecule, and in further embodiments, HLA-B*40: It is a 01 molecule. In another embodiment, the MHC class I molecule is the HLA-B44 molecule. In further embodiments, the HLA-B*44:02 molecule or HLA-B*44:03 molecule Yes. In another embodiment, the MHC class I molecule is an HLA-C03 molecule, further In one embodiment, it is an HLA-C*03:03 molecule. In another embodiment, it is an MHC class I The molecule is the HLA-C04 molecule, and in further embodiments, HLA-C*04:01. It is a child. In another embodiment, the MHC class I molecule is an HLA-C05 molecule, and further In one embodiment, it is an HLA-C*05:01 molecule. In another embodiment, it is an MHC class. The molecule is the HLA-C06 molecule, and in further embodiments, HLA-C06:0 There are two molecules. In another embodiment, the MHC class I molecule is the HLA-C07 molecule. In further embodiments, the HLA-C*07:01 or HLA-C*07:02 molecule In another embodiment, the MHC class I molecule is an HLA-C12 molecule, and further... In the application form, it is an HLA-C*12:03 molecule. In another embodiment, it is an MHC class I molecule. The child is the HLA-C14 molecule, and in further embodiments, the HLA-C*14:02 molecule. That is the case.

[0061] In the embodiment, the tumor antigen peptide is non-covalently bound to an MHC class I molecule (that is, The tumor antigen peptide is loaded into the peptide binding groove / pocket of the MHC class I molecule. (or non-covalently bonded). In another embodiment, the tumor antigen peptide is MHC class It is covalently attached to / bound to molecule I (α chain). In such constructs, tumor antigen peptides and MHC class I molecules (α chains) are typically Short (for example, 5 to 20 residues, preferably about 8 to 12, for example 10) flexible Synthetic fusion protein having a linker or spacer (e.g., polyglycine linker) It is produced as a quality. In another embodiment, the present disclosure is fused to an MHC class I molecule (α chain). This specification provides nucleic acids that encode fusion proteins containing tumor antigen peptides as defined herein. In this embodiment, the MHC class I molecule (α chain)-peptide complex is polymerized. Therefore, in another aspect, the present disclosure relates to the tumor antigen peptide described herein (covalently bonded). It provides a polymer of MHC class I molecules loaded (either covalently or noncovalently). The polymer may be bound to a tag that enables detection of the polymer, such as a fluorescent tag. MH Numerous strategies for the production of MHC multimers, including C dimers, tetramers, pentamers, octamers, etc. It is being developed (Bakker and Schumacher, Current O An overview can be found in *Pinion in Immunology* 2005, 17:428-433. (It is being used.) MHC multimers are useful, for example, for the detection and purification of antigen-specific T cells. Therefore, in another aspect, the present disclosure is specific to tumor antigen peptides as defined herein. Typical CD8 + A method for detecting or purifying (isolating, concentrating) T lymphocytes, Cell populations are loaded (covalently or noncovalently) with tumor antigen peptides Contact with a multimer of MHC class I molecules, and binding by the MHC class I multimer. CD8 + The present invention provides a method for detecting or isolating T lymphocytes. CD8 linked by the Lath I multimer + T lymphocytes can be detected by known methods, such as fluorescence activity. Isolation using Fat-activated cell sorting (FACS) or magnetically activated cell sorting (MACS) That's fine.

[0062] In yet another aspect, the Disclosure may include the nucleic acids, vectors or plastics described herein. Sumid, i.e., a nucleic acid or vector encoding one or more tumor antigen peptides Cells (e.g., host cells), isolated cells are provided in this embodiment. In another embodiment, this The disclosure includes MHC class I cells that are bound to or present the tumor antigen peptide as a result of this disclosure. A molecule expressing a molecule (for example, one of the MHC class I molecules of the alleles disclosed above) Provides cells. In one embodiment, the host cell is a eukaryotic cell, for example, a mammalian cell, preferably is a human cell, cell line, or immortalized cell. In another embodiment, the cell is an antigen-presenting cell. (APC). In one embodiment, the host cells are primary cells, cell lines or immortalized cells. Yes. In another embodiment, the cells are antigen-presenting cells (APCs). The nucleic acids and vectors are It can be introduced into cells via conventional transformation or transfection techniques. The terms "transformation" and "transfection" refer to calcium phosphate or This involves calcium chloride coprecipitation, DEAE-dextran-mediated transfection, and lipofer Cushioning, electroporation, microinjection and virus-borne transfection This refers to a technique for introducing foreign nucleic acids, including cations, into host cells. Alternatively, a preferred method for transfection is, for example, Sambrook et al. This can be found in al. (above) and other laboratory manuals. In vivo feeding Methods for introducing nucleic acids into animal cells are also known, and the vectors disclosed herein are for gene therapy. This can be used to deliver plasmids to their target.

[0063] Using various methods known in the art, one or more cells such as APCs are given Tumor antigen peptides can be loaded. When used herein, tumor antigen peptides "Loading a tumor antigen peptide into a cell" means that the R encoding the tumor antigen peptide is used. NA or DNA is transfected into cells, or alternatively, APCs are transfected into tumors. This means that the cells are transformed with nucleic acids that encode antigen peptides. It can directly bind to MHC class I molecules present in (e.g., peptide pulsed cells). It can be loaded by bringing the exogenous tumor antigen peptide into contact with the cells. The ulcer antigen peptide also has a domain that facilitates its presentation by MHC class I molecules or Motifs, for example, endoplasmic reticulum (ER) retrieval signals, C-terminal Lys-Asp-Glu-Le It can be fused to the u sequence (Wang et al. Eur J Immunol.). See 2004 Dec:34(12):3582-94.

[0064] In another aspect, the present disclosure provides a composition or peptide combination / pool comprising any one or any combination of the tumor antigen peptides (or nucleic acids encoding said peptides) defined herein. In embodiments, the composition comprises any combination (any combination of 2, 3, 4, 5, 6, 7, 8, 9, 10 or more tumor antigen peptides) of the tumor antigen peptides defined herein, or a combination of nucleic acids encoding said tumor antigen peptides. Compositions comprising any combination / partial combination of the tumor antigen peptides defined herein are encompassed by the present disclosure. In another embodiment, the combination or pool may comprise one or more known tumor antigens. In another aspect, the present disclosure provides a composition or peptide combination / pool comprising any one or any combination of the tumor antigen peptides (or nucleic acids encoding said peptides) defined herein. In embodiments, the composition comprises any combination (any combination of 2, 3, 4, 5, 6, 7, 8, 9, 10 or more tumor antigen peptides) of the tumor antigen peptides defined herein, or a combination of nucleic acids encoding said tumor antigen peptides. Compositions comprising any combination / partial combination of the tumor antigen peptides defined herein are encompassed by the present disclosure. In another embodiment, the combination or pool may comprise one or more known tumor antigens. In another aspect, the present disclosure provides a composition or peptide combination / pool comprising any one or any combination of the tumor antigen peptides (or nucleic acids encoding said peptides) defined herein. In embodiments, the composition comprises any combination (any combination of 2, 3, 4, 5, 6, 7, 8, 9, 10 or more tumor antigen peptides) of the tumor antigen peptides defined herein, or a combination of nucleic acids encoding said tumor antigen peptides. Compositions comprising any combination / partial combination of the tumor antigen peptides defined herein are encompassed by the present disclosure. In another embodiment, the combination or pool may comprise one or more known tumor antigens. In another aspect, the present disclosure provides a composition or peptide combination / pool comprising any one or any combination of the tumor antigen peptides (or nucleic acids encoding said peptides) defined herein. In embodiments, the composition comprises any combination (any combination of 2, 3, 4, 5, 6, 7, 8, 9, 10 or more tumor antigen peptides) of the tumor antigen peptides defined herein, or a combination of nucleic acids encoding said tumor antigen peptides. Compositions comprising any combination / partial combination of the tumor antigen peptides defined herein are encompassed by the present disclosure. In another embodiment, the combination or pool may comprise one or more known tumor antigens. In another aspect, the present disclosure provides a composition or peptide combination / pool comprising any one or any combination of the tumor antigen peptides (or nucleic acids encoding said peptides) defined herein. In embodiments, the composition comprises any combination (any combination of 2, 3, 4, 5, 6, 7, 8, 9, 10 or more tumor antigen peptides) of the tumor antigen peptides defined herein, or a combination of nucleic acids encoding said tumor antigen peptides. Compositions comprising any combination / partial combination of the tumor antigen peptides defined herein are encompassed by the present disclosure. In another embodiment, the combination or pool may comprise one or more known tumor antigens. In another aspect, the present disclosure provides a composition or peptide combination / pool comprising any one or any combination of the tumor antigen peptides (or nucleic acids encoding said peptides) defined herein. In embodiments, the composition comprises any combination (any combination of 2, 3, 4, 5, 6, 7, 8, 9, 10 or more tumor antigen peptides) of the tumor antigen peptides defined herein, or a combination of nucleic acids encoding said tumor antigen peptides. Compositions comprising any combination / partial combination of the tumor antigen peptides defined herein are encompassed by the present disclosure. In another embodiment, the combination or pool may comprise one or more known tumor antigens. In another aspect, the present disclosure provides a composition or peptide combination / pool comprising any one or any combination of the tumor antigen peptides (or nucleic acids encoding said peptides) defined herein. In embodiments, the composition comprises any combination (any combination of 2, 3, 4, 5, 6, 7, 8, 9, 10 or more tumor antigen peptides) of the tumor antigen peptides defined herein, or a combination of nucleic acids encoding said tumor antigen peptides. Compositions comprising any combination / partial combination of the tumor antigen peptides defined herein are encompassed by the present disclosure. In another embodiment, the combination or pool may comprise one or more known tumor antigens. In another aspect, the present disclosure provides a composition or peptide combination / pool comprising any one or any combination of the tumor antigen peptides (or nucleic acids encoding said peptides) defined herein. In embodiments, the composition comprises any combination (any combination of 2, 3, 4, 5, 6, 7, 8, 9, 10 or more tumor antigen peptides) of the tumor antigen peptides defined herein, or a combination of nucleic acids encoding said tumor antigen peptides. Compositions comprising any combination / partial combination of the tumor antigen peptides defined herein are encompassed by the present disclosure. In another embodiment, the combination or pool may comprise one or more known tumor antigens.

[0065] Thus, in another aspect, the present disclosure provides a composition comprising any one or any combination of the tumor antigen peptides defined herein and a cell expressing an MHC class I molecule (e.g., an MHC class I molecule of one of the alleles disclosed above). APCs for use in the present disclosure are not limited to a particular type of cell and include professional APCs such as dendritic cells (DCs), Langerhans cells, macrophages, and B cells that are known to present proteinaceous antigens on their cell surface such that they are recognized by CD8 T lymphocytes. For example, APCs can be obtained by inducing DCs from peripheral blood mononuclear cells in vitro, ex vivo or in vivo and then contacting (stimulating) them with tumor antigen peptides. APCs can also be the Thus, in another aspect, the present disclosure provides a composition comprising any one or any combination of the tumor antigen peptides defined herein and a cell expressing an MHC class I molecule (e.g., an MHC class I molecule of one of the alleles disclosed above). APCs for use in the present disclosure are not limited to a particular type of cell and include professional APCs such as dendritic cells (DCs), Langerhans cells, macrophages, and B cells that are known to present proteinaceous antigens on their cell surface such that they are recognized by CD8 T lymphocytes. For example, APCs can be obtained by inducing DCs from peripheral blood mononuclear cells in vitro, ex vivo or in vivo and then contacting (stimulating) them with tumor antigen peptides. APCs can also be the Thus, in another aspect, the present disclosure provides a composition comprising any one or any combination of the tumor antigen peptides defined herein and a cell expressing an MHC class I molecule (e.g., an MHC class I molecule of one of the alleles disclosed above). APCs for use in the present disclosure are not limited to a particular type of cell and include professional APCs such as dendritic cells (DCs), Langerhans cells, macrophages, and B cells that are known to present proteinaceous antigens on their cell surface such that they are recognized by CD8 T lymphocytes. For example, APCs can be obtained by inducing DCs from peripheral blood mononuclear cells in vitro, ex vivo or in vivo and then contacting (stimulating) them with tumor antigen peptides. APCs can also be the Thus, in another aspect, the present disclosure provides a composition comprising any one or any combination of the tumor antigen peptides defined herein and a cell expressing an MHC class I molecule (e.g., an MHC class I molecule of one of the alleles disclosed above). APCs for use in the present disclosure are not limited to a particular type of cell and include professional APCs such as dendritic cells (DCs), Langerhans cells, macrophages, and B cells that are known to present proteinaceous antigens on their cell surface such that they are recognized by CD8 T lymphocytes. For example, APCs can be obtained by inducing DCs from peripheral blood mononuclear cells in vitro, ex vivo or in vivo and then contacting (stimulating) them with tumor antigen peptides. APCs can also be the CD8 + Thus, in another aspect, the present disclosure provides a composition comprising any one or any combination of the tumor antigen peptides defined herein and a cell expressing an MHC class I molecule (e.g., an MHC class I molecule of one of the alleles disclosed above). APCs for use in the present disclosure are not limited to a particular type of cell and include professional APCs such as dendritic cells (DCs), Langerhans cells, macrophages, and B cells that are known to present proteinaceous antigens on their cell surface such that they are recognized by CD8 T lymphocytes. For example, APCs can be obtained by inducing DCs from peripheral blood mononuclear cells in vitro, ex vivo or in vivo and then contacting (stimulating) them with tumor antigen peptides. APCs can also be the Thus, in another aspect, the present disclosure provides a composition comprising any one or any combination of the tumor antigen peptides defined herein and a cell expressing an MHC class I molecule (e.g., an MHC class I molecule of one of the alleles disclosed above). APCs for use in the present disclosure are not limited to a particular type of cell and include professional APCs such as dendritic cells (DCs), Langerhans cells, macrophages, and B cells that are known to present proteinaceous antigens on their cell surface such that they are recognized by CD8 T lymphocytes. For example, APCs can be obtained by inducing DCs from peripheral blood mononuclear cells in vitro, ex vivo or in vivo and then contacting (stimulating) them with tumor antigen peptides. APCs can also be the Thus, in another aspect, the present disclosure provides a composition comprising any one or any combination of the tumor antigen peptides defined herein and a cell expressing an MHC class I molecule (e.g., an MHC class I molecule of one of the alleles disclosed above). APCs for use in the present disclosure are not limited to a particular type of cell and include professional APCs such as dendritic cells (DCs), Langerhans cells, macrophages, and B cells that are known to present proteinaceous antigens on their cell surface such that they are recognized by CD8 T lymphocytes. For example, APCs can be obtained by inducing DCs from peripheral blood mononuclear cells in vitro, ex vivo or in vivo and then contacting (stimulating) them with tumor antigen peptides. APCs can also be the Thus, in another aspect, the present disclosure provides a composition comprising any one or any combination of the tumor antigen peptides defined herein and a cell expressing an MHC class I molecule (e.g., an MHC class I molecule of one of the alleles disclosed above). APCs for use in the present disclosure are not limited to a particular type of cell and include professional APCs such as dendritic cells (DCs), Langerhans cells, macrophages, and B cells that are known to present proteinaceous antigens on their cell surface such that they are recognized by CD8 T lymphocytes. For example, APCs can be obtained by inducing DCs from peripheral blood mononuclear cells in vitro, ex vivo or in vivo and then contacting (stimulating) them with tumor antigen peptides. APCs can also be the Thus, in another aspect, the present disclosure provides a composition comprising any one or any combination of the tumor antigen peptides defined herein and a cell expressing an MHC class I molecule (e.g., an MHC class I molecule of one of the alleles disclosed above). APCs for use in the present disclosure are not limited to a particular type of cell and include professional APCs such as dendritic cells (DCs), Langerhans cells, macrophages, and B cells that are known to present proteinaceous antigens on their cell surface such that they are recognized by CD8 T lymphocytes. For example, APCs can be obtained by inducing DCs from peripheral blood mononuclear cells in vitro, ex vivo or in vivo and then contacting (stimulating) them with tumor antigen peptides. APCs can also be the One or more of the tumor antigen peptides shown are administered to a subject and present the tumor antigen peptide APC is induced in the subject's body and activated to present the tumor antigen peptide in vivo It can be. The phrases "inducing APC" or "stimulating APC" refer to one or more tumors of a cell such that the tumor antigen peptide The peptide is presented on its surface by MHC class I molecules, Contacting the antigen peptide, or a nucleic acid encoding the tumor antigen peptide, or loading them As described herein, according to the present disclosure, the tumor antigen peptide Peptides may be indirectly loaded using longer peptides / polypeptides containing, for example, the sequence of the tumor antigen peptide (including native proteins), and then the tumor antigen peptide / The MHC class I complex is processed within the APC (e.g., By proteases) to generate on the cell surface. After loading the APC with the tumor antigen peptide and enabling the APC to present the tumor antigen peptide The APC can be administered to the subject as a vaccine . For example, ex vivo administration can include the following steps: (a) harvesting APCs from a first subject And (b) contacting / loading the APCs of step (a) with the tumor antigen peptide to form MHC class I / tumor antigen peptide complexes on the surface of the APC And (c) administering the peptide-loaded APCs to a second subject who requires treatment . The first subject and the second subject may be the same subject (e.g., an autologous vaccine) Or different subjects (e.g., an allogeneic vaccine). Alternatively, according to the present disclosure

[0066] The first subject and the second subject may be the same subject (e.g., an autologous vaccine), Or different subjects (e.g., an allogeneic vaccine). Alternatively, according to the present disclosure A composition for inducing antigen-presenting cells (e.g., a pharmaceutical composition) for manufacturing The use of tumor antigen peptides (or combinations thereof) as described in the details is provided. In addition Therefore, this disclosure relates to a method or process for producing a pharmaceutical composition for inducing antigen-presenting cells. The method or process provides a drug containing tumor antigen peptides, or a combination thereof. The process includes mixing or formulating with a scientifically acceptable carrier. Tumor anti- MHC class I cells loaded with one or any combination of the proto-peptides Molecules (e.g., HLA-A1, HLA-A2, HLA-A3, HLA-A11, HLA- A24, HLA-A25, HLA-A29, HLA-A32, HLA-B07, HLA- B08, HLA-B14, HLA-B15, HLA-B18, HLA-B39, HLA- B40, HLA-B44, HLA-C03, HLA-C04, HLA-C05, HLA- AP expressing HLA-C06, HLA-C07, HLA-C12, or HLA-C14 molecules Cells such as C are CD8 + T lymphocytes, for example, autologous CD8 + Stimulate / amplify T lymphocytes It can be used for that purpose. Therefore, in another aspect, this disclosure is defined herein Among the tumor antigen peptides (or nucleic acids or vectors that encode them) Any one of the following, or any combination: MHC class I molecules and T lymphocytes, more specific CD8 + Cells that express T lymphocytes (e.g., CD8 + A group of cells including T lymphocytes The present invention provides a composition containing ).

[0067] In embodiments, the composition includes a buffer, an excipient, a carrier, a diluent, and / or a culture medium (e.g.) In further embodiments, the mixture may include a culture medium. In more embodiments, it may also include a buffer, an excipient, a carrier, a diluent, and / or culture media are pharmaceutically acceptable buffers, excipients, carriers, diluents and / or Culture medium (culture medium). When used herein, "pharmaceutically acceptable buffer, excipient" The carrier, diluent and / or culture medium are physiologically compatible and the biological activity of the active ingredient. Any and all solvents, buffers, and binders that do not interfere with the effectiveness of the substance and are non-toxic to the target substance. Lubricants, fillers, thickeners, disintegrants, plasticizers, coating agents, barrier layer formulations, lubricants Stabilizers, release retarders, dispersion media, coating agents, antibacterial and antifungal agents, isotonic agents, etc. This includes. Using such culture media and drugs with pharmaceutically active substances is a technical matter. It is well known in the field (Rowe et al., Handbook of pharmacy) aceutical excipients, 2003, 4 th edition, Ph Armaceutical Press, London, UK). Any conventional culture medium or Unless the drug is incompatible with the active compound (peptide, cell), the composition of this disclosure is not applicable. The use of these is intended. In embodiments, buffers, excipients, carriers, and / or Culture media are buffers, excipients, carriers, and / or culture media that do not exist in nature. In this state, a tumor antigen peptide as defined herein, or one or more such tumor antigen peptides. One or more nucleic acids (e.g., mRNA) that code for a liposome, for example, catkins It is contained within or complexed with liposomes (e.g., Vit or MT et al.,Recent Pat Drug Deliv Formu See l.2013 Aug;7(2):99-110.

[0068] In another aspect, the present disclosure provides a composition comprising any one or any combination of the tumor antigen peptides (or nucleic acids encoding said peptides) defined herein, and one or more of a buffer, excipient, carrier, diluent, and / or medium. For compositions comprising cells (e.g., APCs, T lymphocytes), the composition comprises a suitable medium that allows for maintenance of viable cells. Representative examples of such media include saline, Earl’s balanced salt solution (Life Technologies®), and also PlasmaLyte® (Baxter International®). In embodiments, the composition (e.g., a pharmaceutical composition) is an “immunogenic composition”, “vaccine composition” or a “vaccine”. As used herein, the terms “immunogenic composition”, “vaccine composition” or “vaccine” refer to a composition or formulation that contains one or more tumor antigen peptides or vaccine vectors and is capable of inducing an immune response against one or more tumor antigen peptides present therein when administered to a subject. Vaccination methods for inducing an immune response in mammals include the use of a vaccine or vaccine vector administered by any conventional route known in the vaccine art, e.g., via the mucosal (e.g., ocular, intranasal, pulmonary, oral, gastric, intestinal, rectal, vaginal, or urinary tract) surface, parenterally (e.g., subcutaneously, intradermally, intramuscularly, intravenously, or intraperitoneally), or by topical administration (e.g., via a transdermal delivery system such as a patch). In embodiments, the tumor antigen peptides (or combinations thereof) are conjugated to a carrier protein to increase the immunogenicity of the tumor antigen peptides (conjugate vaccine). (Baxter International®). In embodiments, the composition (e.g., a pharmaceutical composition) is an “immunogenic composition”, “vaccine composition” or a “vaccine”. As used herein, the terms “immunogenic composition”, “vaccine composition” or “vaccine” refer to a composition or formulation that contains one or more tumor antigen peptides or vaccine vectors and is capable of inducing an immune response against one or more tumor antigen peptides present therein when administered to a subject. Vaccination methods for inducing an immune response in mammals include the use of a vaccine or vaccine vector administered by any conventional route known in the vaccine art, e.g., via the mucosal (e.g., ocular, intranasal, pulmonary, oral, gastric, intestinal, rectal, vaginal, or urinary tract) surface, parenterally (e.g., subcutaneously, intradermally, intramuscularly, intravenously, or intraperitoneally), or by topical administration (e.g., via a transdermal delivery system such as a patch). In embodiments, the tumor antigen peptides (or combinations thereof) are conjugated to a carrier protein [[ID=ip>20]]to increase the immunogenicity of the tumor antigen peptides (conjugate vaccine). (conjugate vaccine). The vaccination method for inducing an immune response in mammals includes the use of a vaccine or vaccine vector administered by any conventional route known in the vaccine field, for example, through the mucosal (for example, eye, intranasal, lung, oral, stomach, intestine, rectum, vagina, or urinary tract) surface, parenterally (for example, subcutaneous, intradermal, intramuscular, intravenous, or intraperitoneal) route, or by topical administration (for example, through a transdermal delivery system such as a patch). In embodiments, the tumor antigen peptides (or combinations thereof) are conjugated to a carrier protein to increase the immunogenicity of the tumor antigen peptides (conjugate vaccine). (conjugate vaccine). The vaccination method for inducing an immune response in mammals includes the use of a vaccine or vaccine vector administered by any conventional route known in the vaccine field, for example, through the mucosal (for example, eye, intranasal, lung, oral, stomach, intestine, rectum, vagina, or urinary tract) surface, parenterally (for example, subcutaneous, intradermal, intramuscular, intravenous, or intraperitoneal) route, or by topical administration (for example, through a transdermal delivery system such as a patch). In embodiments, the tumor antigen peptides (or combinations thereof) are conjugated to a carrier protein to increase the immunogenicity of the tumor antigen peptides (conjugate vaccine). Therefore, this disclosure refers to tumor antigen peptides (or combinations thereof), or Nucleic acids encoding tumor antigen peptides or combinations thereof, and carrier proteins. To provide a composition (conjugate). For example, a tumor antigen peptide or nucleic acid is provided. LL-like receptor (TLR) ligands (e.g., Zom et al., Adv Immun) ol.2012,114:177-201) or polymer / dendrimer (e.g., L iu et al., Biomacromolecules.2013 Aug 12; Conjugate or complex into (see 14(8):2798-806) Alternatively, the immunogenic composition or vaccine may further comprise an adjuvant. "Adjuvant" refers to an antigen (tumor antigen peptide, nucleic acid and / or cell as defined in this disclosure). When added to immunogenic agents such as ), the immune response to the drug in the host upon exposure to the mixture is not This refers to a substance that specifically improves or enhances something. Adjuvants are currently used in the field of vaccines. Examples of the product include (1) mineral salts (such as aluminum phosphate and aluminum hydroxide). (1) Aluminum salt, calcium phosphate gel, squalene, (2) oil-based adjuvant (For example, oil emulsions and surfactant-based formulations), MF59 (micro Fluidized detergent (stabilized oil-in-water emulsion), QS21 (purified saponin), AS02 [SB AS2] (Oil-in-water emulsion + MPL + QS-21), (3) Particulate adjuvant, For example, virosom (a monolayer liposome incorporating influenza hemagglutinin) (Lu), AS04 (containing MPL [SBAS4] aluminum salt), ISCOMS (Saponin (4) Microbial induction (a structural complex of ions and lipids), polylactide coglycol (PLG), Body (natural and synthetic), e.g., monophosphoryl lipid A (MPL), Detox (MPL) L+M.Phlei cell wall skeleton), AGP[RC-529] (synthetic acylated monosaccharide), D C_Chol (a lipoid immunostimulant that can self-construct into liposomes), OM -174 (Lipid A derivative), CpG motif (containing immunostimulatory CpG motif) (Constituting oligonucleotides), modified LT and CT (to provide non-toxic adjuvant effects) (5) Genetically modified bacterial toxin immunoassays), (6) Endogenous human immunomodulators, for example, hG M-CSF or hIL-12 (either a protein or an encoded plasmid) Cytokines that can be administered as follows: Immudaptin (C3d tandem array), (6) Examples include an inert vehicle such as gold particles.

[0069] In the embodiment, the tumor antigen peptide or a composition containing the same is in a lyophilized form. In another embodiment, the tumor antigen peptide or a composition containing it is a liquid composition. In further embodiments, the tumor antigen peptide is present in the composition at a concentration of about 0.01 μg / mL to about 1 The concentration is 00 μg / mL. In further embodiments, the tumor antigen peptide is present in the composition. Approx. 0.2 μg / mL ~ approx. 50 μg / mL, approx. 0.5 μg / mL ~ approx. 10, 20, 30, 4 0, or 50 μg / mL, approximately 1 μg / mL to approximately 10 μg / mL, or approximately 2 μg / mL This is the concentration.

[0070] Any of the tumor antigen peptides as defined herein, as described herein Load or combine one or any combination of MHC Class I Cells such as APCs that express molecules can be used to target CD8+ T lymphocytes in vivo or ex vivo. It may be used to stimulate / amplify. Therefore, in another aspect, this disclosure is used herein It can interact with or bind to the MHC class I molecule / tumor antigen peptide complex described. T cell receptors (TCRs), and nucleic acid molecules that encode such TCR molecules, The present disclosure provides a vector containing such nucleic acid molecules. The TCR according to this disclosure is preferably In vitro or in vivo, on the surface of living cells, it is loaded onto MHC class I molecules, Alternatively, it specifically interacts with tumor antigen peptides presented by MHC class I molecules. It can bind. The nucleic acid encoding the TCR, in particular the TCR of this disclosure, is, for example, M Novel T lymphocyte clones that specifically recognize HC class I / tumor antigen peptide complexes To generate, T lymphocytes (e.g., CD8 + T lymphocytes) or other types of lymphocytes It can be applied to genetically transform / modify. In certain embodiments, obtained from a patient T lymphocytes (e.g., CD8) + T lymphocytes recognize one or more tumor antigen peptides. The cells are transformed to express TCR, and the transformed cells are administered to the patient (autologous). Cell transfusion). In certain embodiments, T lymphocytes (e.g., CD8) obtained from a donor are used. + T Lymphocytes are transformed to express one or more TCRs that recognize tumor antigen peptides. The transformed cells are then administered to the recipient (allogeneic cell transfusion). Another implementation In this context, the present disclosure relates to T lymphocytes, for example, those encoding tumor antigen peptide-specific TCRs. CD8 transformed / transfected by a processor or plasmid + T lymphocytes This provides. In further embodiments, this disclosure provides a tumor antigen peptide-specific TCR for transformation. The present invention provides a method for treating patients using replaced autologous or allogeneic cells. In this state, tumor antigen-specific TCRs in the production of autologous or allogeneic cells for cancer treatment Use is provided.

[0071] In some embodiments, a patient treated with a composition of the present disclosure (e.g., a pharmaceutical composition) Treatment involves allogeneic stem cell transplantation (ASCL), allogeneic lymphocyte infusion, or autologous lymphocyte infusion. It is treated before or after. The compositions of this disclosure are ex vivo against tumor antigen peptides. Activated allogeneic T lymphocytes (e.g., CD8) + T lymphocytes, loaded with tumor antigen peptides Allogeneic or autologous APC vaccines, tumor antigen peptide vaccines, and tumor antigen-specific vaccines. Allogeneic or autologous T lymphocytes transformed at the TCR (e.g., CD8) + T lymphocytes) This includes lymphocytes. The T lymphocytes that can recognize tumor antigen peptides according to this disclosure The method for providing the loan is to provide the target (e.g., graft recipient), e.g., ASC In T and / or donor lymphocyte infusion (DLI) recipients, tumor antigen peptides It may be produced for tumor cells that express the gene, and it can be specifically targeted. Therefore, this disclosure specifically recognizes tumor antigen peptide / MHC class I molecule complexes. CD8, which encodes and expresses a T cell receptor that can recognize or bind to it. + T lymphocytes To provide the T lymphocyte (e.g., CD8) + T lymphocytes are recombinant (manipulated) These may be naturally selected T lymphocytes. Therefore, this specification applies to in vitro or This can be done in vivo (i.e., APC is loaded with tumor antigen peptides) This procedure was performed in patients who received thin or in patients who received tumor antigen peptides. Under conditions favorable to induce T cell activation and proliferation, undifferentiated lymphocytes can be... Tumor antigen peptide / MHC class I molecule complex (typically, on the surface of cells such as APCs) The present disclosure includes a step of contacting (the expressed) CD8 + A small amount of T lymphocytes are produced. At the very least, it provides two methods: a combination of tumor antigen peptides bound to MHC class I molecules. Population C, which can recognize multiple tumor antigen peptides using combination or pooling. D8 + It is possible to generate T lymphocytes, or tumor antigen-specific or targeted T lymphocytes. T lymphocytes are MHC class I molecule / tumor antigen peptide complexes (i.e., manipulated or is a recombinant CD8 + TCRs (more specifically, α-chain and By cloning one or more nucleic acids (genes) that encode the β chain, It may be produced / generated in vitro or ex vivo. Tumor antigen peptide-specific T The nucleic acid encoding CR is obtained using methods known in the relevant field of art, and is a tumor antigen peptide. Ex vivo activated T lymphocytes (for example, loaded with tumor antigen peptides) against the thrombocytopenia Obtained from individuals that exhibit an immune response to APCs or peptide / MHC molecular complexes. The tumor antigen peptide-specific TCRs of this disclosure can be used in host cells and / or graft cells. Recombinant expression occurs in host lymphocytes obtained from septics or graft donors. They can be arbitrarily differentiated in vitro and provide cytotoxic T lymphocytes (CTLs). Nucleic acids encoding α and β chains (transgenes) undergo transfection (e.g., electrical Any suitable method such as perforation or transduction (e.g., using a viral vector) It may be used to introduce T cells (for example, from the subject being treated or another individual). Modified CD8 expressing a TCR specific to tumor antigen peptides + T lymphocytes are well known It can be grown in vitro using culture methods.

[0072] This disclosure relates to tumor antigen peptides (i.e., MHC class I molecules expressed on the cell surface). Specific induction by the bound tumor antigen peptide, or by a combination of tumor antigen peptides. Isolated CD8 is guided, activated, and / or amplified (proliferated). + T-rin This disclosure provides pocytes. This disclosure also provides tumor antigen peptides, or combinations thereof. CD8 can recognize the match + T lymphocytes (that is, MHC class I molecules) The present invention provides one or more combined tumor antigen peptides and compositions comprising the tumor antigen peptides. In another aspect, this disclosure relates to one or more MHC class I molecules / tumor antigens described herein. CD8 specifically recognizes peptide complexes. + Cell population or cell culture enriched with T lymphocytes Nutrients (for example, CD8) + This specification provides a population of T lymphocytes. MH loaded with one or more of the tumor antigen peptides disclosed in the book (e.g., presented) Using cells such as APCs that express Class C I molecules, specific T lymphocytes are subjected to ex vivo testing. It can be obtained by carrying out propagation. The term "concentrated" as used herein refers to , tumor antigen-specific CD8 in the population + The proportion of T lymphocytes compared to the natural population of cells That is, more pronounced than those not subjected to the ex vivo proliferation process of specific T lymphocytes. This means that the tumor antigen peptide specificity in a cell population target CD8 + The percentage of T lymphocytes is at least about 0.5%, for example, at least about 0.6%. These are 0.7%, 0.8%, 0.9%, 1%, 1.5%, 2%, or 3%. In this embodiment, tumor antigen peptide-specific CD8 in a cell population + The percentage of T lymphocytes is approximately 0 0.5-10%, 0.5-8%, 0.5-5%, 0.5-4%, 0.5- Approximately 3%, approximately 1% to approximately 5%, approximately 1% to approximately 4%, approximately 1% to approximately 3%, approximately 2% to approximately 5%, approximately 2% to The percentages are approximately 4%, 2% to 3%, 3% to 5%, or 3% to 4%. One or more objectives. CD specifically recognizes MHC class I molecule / peptide (tumor antigen peptide) complexes. A cell population or culture enriched with 8+ T lymphocytes (e.g., CD8+ T lymphocyte population) The group may be used in tumor antigen-based cancer immunotherapy, as detailed below. In some embodiments, tumor antigen peptide-specific CD8 + A population of T lymphocytes, for example, The tumor antigen peptides as defined herein are loaded (covalently or noncovalently). The resulting MHC class I molecule is further enriched using a polymer. Therefore, this disclosure For example, tumor antigen peptide-specific CD8 + The proportion of T lymphocytes is at least about 30%. 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, Tumor antigen peptide-specific CD8 is 98%, 99%, or 100%. + T lymphocyte sperm Provides a manufactured or isolated population.

[0073] This disclosure further relates to any of the following: The tumor antigen peptide, nucleic acid, expression vector, T cell receptor, cell (e.g., T lymphocyte) The use of , APC), and / or compositions, or any combination thereof. In some embodiments, the pharmaceutical product is for the treatment of cancer, and is, for example, a cancer vaccine. This disclosure is for use, for example, as a cancer vaccine for the treatment of cancer, and is not subject to any discretion as described herein. Tumor antigen peptides, nucleic acids, expression vectors, T cell receptors, cells (e.g., T lymphocytes) APCs), and / or compositions (e.g., vaccine compositions), or any combination thereof. Regarding combinations. The tumor antigen peptide sequences identified herein are i) injected into tumor patients. Used for in vitro priming and proliferation of tumor antigen-specific T cells, and / or ii) to induce or enhance the antitumor T cell response in cancer patients It can be used as cutin, or for the production of synthetic peptides.

[0074] In another aspect, the present disclosure describes the present specification as a vaccine for treating cancer in the subject. Tumor antigen peptides described in the book, or combinations thereof (e.g., peptide pools) This disclosure also provides the use of ) as a vaccine for treating cancer in the subject. , tumor antigen peptides described herein, or combinations thereof (e.g., peptide The use of dopure is provided. In the embodiment, the target is tumor antigen peptide-specific CD8 + The recipient is a T lymphocyte. Therefore, in another embodiment, this disclosure treats cancer. The method of providing (for example, reducing the number of tumor cells, killing tumor cells) The method involves an effective amount of one or more MHC class I molecule / tumor antigen peptide complexes (AP). Recognizes (i.e., expresses a TCR that binds to) cells such as C expressed on the cell surface. (do) ​​CD8 + This includes administering (injecting) T lymphocytes to a target that needs them. In the embodiment, the method is the CD8 + After administration / infusion of T lymphocytes, an effective amount of tumor antigen M loaded with peptides, or combinations thereof, and / or tumor antigen peptides Cells expressing HC class I molecules (e.g., APCs such as dendritic cells) are administered to the subject. This further includes the following. In yet another embodiment, the method involves a therapeutically effective amount of one or more tumor antibodies This includes administering dendritic cells loaded with the proto-peptide to a target that requires it. In one embodiment, the method involves an effective amount of tumor antimicrobial agent (MHC class I) present by an MHC class I molecule. Allogeneic or autologous cells expressing recombinant TCRs that bind to the proto-peptide, which require This includes administering the drug to patients.

[0075] In another aspect, the disclosure describes a treatment for the cancer in question (e.g., reducing the number of tumor cells). Loaded with tumor antigen peptides, or combinations thereof, to kill tumor cells. CD8 recognizes one or more MHC class I molecules (to present) + The use of T lymphocytes is proposed. In another aspect, the Disclosure provides a method for treating the cancer in question (e.g., reducing the number of tumor cells). Tumor antigen peptides for the preparation / manufacturing of pharmaceuticals (to kill tumor cells), Alternatively, it recognizes one or more MHC class I molecules loaded (presenting) with a combination of these. CD8 + The use of T lymphocytes is provided. In another aspect, the disclosure provides treatment for the target cancer. For use in (for example, to reduce the number of tumor cells and kill tumor cells) One or more loaded (presenting) tumor antigen peptides, or combinations thereof. CD8 recognizes MHC class I molecules. + Provides T lymphocytes (cytotoxic T lymphocytes). In further embodiments, use involves the use of tumor antigen peptide-specific CD8+ T lymphocytes. Subsequently, an effective amount of tumor antigen peptide (or a combination thereof), and / or tumor Cells expressing one or more MHC class I molecules loaded (presenting) ulcer antigen peptides (for example) This further includes the use of APC.

[0076] This disclosure also relates to any or any combination thereof of the tumor antigen peptides disclosed herein. The target is the immune response against tumor cells expressing human class I MHC molecules loaded with a specific compound. The present invention provides a method for generating a tumor antigen peptide or tumor antigen peptide Cytotoxic T lymphocytes that specifically recognize class I MHC molecules loaded with a combination This disclosure also includes loading tumor antigen peptides or combinations thereof. To induce an immune response against tumor cells expressing human class I MHC molecules any of the tumor antigen peptides or combinations of tumor antigen peptides disclosed herein. The use of cytotoxic T lymphocytes that specifically recognize class I MHC molecules loaded with these molecules is also proposed. To provide.

[0077] In embodiments, the methods or uses described herein are performed by the patient before treatment / use. Determining the HLA class I alleles expressed and the HLA class I alleles expressed by the patient Administering or using tumor antigen peptides that bind to one or more of the LASS I alleles. This further includes, for example, a patient with HLA-A2*01, HLA-B14*01 and If HLA-C05*01 is determined to be expressed, (i) SEQ ID NOs. 14, 17, 45, 48, 51, 56, 75, 77, 82, 98 and / or 100 (HLA-A2*01) (ii) binds to (ii) Sequence ID 53 (binds to HLA-B14*01), and / ma (iii) Tumor antigen peptide of sequence number 27 (which binds to HLA-C05*01) Any combination may be administered to or used by the patient.

[0078] In the embodiment, the cancer is a solid tumor, preferably ovarian cancer. This refers to ovarian cancer. In this embodiment, ovarian cancer includes epithelial cancer, serous cancer, and small cell carcinoma. Primary peritoneal cancer, clear cell carcinoma or adenocarcinoma, endometrial adenocarcinoma, malignant mixed Müller tumor Mucinous gland carcinoma or cystadenocarcinoma, malignant Brenner tumor, transitional cell carcinoma, sex cord-stromal tumor, granuloma Granular cell tumor, Sertoli-Leydig tumor, germ cell tumor, undifferentiated germ cell tumor, choriocarcinoma, Immature (solid) teratoma or mature teratoma, yolk sac tumor, squamous cell carcinoma, or secondary It is ovarian cancer. In this embodiment, the ovarian cancer is type I ovarian carcinoma. In another embodiment, In this embodiment, the ovarian cancer is type II ovarian cancer. In further embodiments, serous carcinoma is high-grade serous carcinoma (HGSC). In this context, ovarian cancer is classified as stage I, II, III, or IV ovarian cancer.

[0079] In embodiments, tumor antigen peptides, nucleic acids, expression vectors, T cell receptors, etc., according to the present disclosure Cells (e.g., T lymphocytes, APCs), and / or compositions, or any combination thereof. Combinations include chemotherapy (e.g., vinca alkaloids, drugs that inhibit microtubule formation (e.g., Colchicine and its derivatives), anti-angiogenic agents, therapeutic antibodies, EGFR targeting agents, tyrofoam Syn-kinase targeting agents (e.g., tyrosine kinase inhibitors), transition metal complexes, proteaso Antibodies, antimetabolites (e.g., nucleoside analogs), alkylating agents, platinum-based drugs, Anthracycline antibiotics, topoisomerase inhibitors, macrolides, retinoids (examples) For example, all trans-retinoic acid or its derivatives), gardamine or its derivatives (17-AAG), immune checkpoint inhibitors (e.g., PD-1 / -PD-L1 inhibitors) Agents and CTLA-4 inhibitors, B7-1 / B7-2 inhibitors) antibodies, cell-based therapies (e.g.) For example, to treat cancer, such as CAR T cells, one or more additional active drugs or It may be used in combination with therapy. In embodiments, the tumor antigen peptide according to the present disclosure Nucleic acids, expression vectors, T cell receptors, cells (e.g., T lymphocytes, APCs), and / or These compositions are administered / used in combination with immune checkpoint inhibitors. [Examples]

[0080] This disclosure is illustrated in further detail by the following non-limiting embodiments.

[0081] Example 1: Materials and Method Human HGSC samples. Tumor fragments of HGSC1-6 and matched normal adjacent cells of HGSC1-3. The contact information was obtained from Tissue Solutions (Glasgow, GB). Tumor tissue (OV606) or ascites (OV633 and OV642) are Princes s Margaret Cancer Registry(Toronto, ON, Ca Obtained from nada. The snap-frozen sample was subjected to RNA extraction. It was used for the isolation of MHCI-related peptides. RNA sequencing of OvCa48-114. The data was collected under the project name PRJNA398141 by the National Ce ter for Biotechnology Information Seque Download from nce Read Archive and convert to a fastq file. The samples were processed in the same way as other samples. The raw MS data of the samples in this cohort were identified as PXD00. ProteomeXchange Consortium via PRIDE Partner 7635 Downloaded from [website / platform name]. HLA typing for each sample was done using the default parameter (24). RNA sequencing using OptiType(trademark) v1.0 (RNA-S The data was obtained from the eq) data. Sample information is presented in Table 1. [Table 1]

[0082] RNA extraction and sequencing. For HGSC1-6, use AllPrep(registered trademark). Using the DNA / RNA / miRNA Universal Kit (Qiagen), manufacturers Total RNA was isolated according to the recommendations of OV606, OV633, and OV642. Then, we used TRIzol(registered trademark) (Invitrogen) to isolate total RNA. The RNA of each sample was analyzed using 2100 Bioanalyzer(registered trademark)(Agilent). We evaluated it using Genomics and confirmed that RIN > 6, and used that to determine the sample. One RNA-Seq replication was performed per sample. The KAPA standard mRNA-Seq Kit was used. Using this method, a cDNA library was prepared from polyA-rich mRNA. Further amplification is achieved using HiSeq(registered trademark) 2000 or Illumina NextSe Used for paired-end RNA-Seq with q(registered trademark)500, yielding 105 million per sample. We gained a lead of 0 to 300 million.

[0083] Generation of a customized reference database for MS analysis. For each sample, The customized "Global Cancer Database" is, as mentioned above (15 and US provisional Application No. 62 / 724,760), "Canonical cancer proteome" and "cancer-specific proteome" It was generated by linking two modules of "roteome". In short, T Using rimmomatic v0.35(25), RNA-Seq reads are added The pters and low-quality 3' bases were trimmed. This generates a canonical cancer proteome. Therefore, the trimmed lead is referenced using STAR v2.5.1b. Aligned to version GRCh38.88. Transcript expression was set to default parameters. Using the method, with kallisto v0.43.0, the number of transcripts per million ( Quantitative analysis was performed using tpm. Nucleotide variants were analyzed using FreeBayes(26) Identify and use agnostic as input to pyGeno(27) ) Converted to single nucleotide polymorphism file format. Then, sample-specific proteo for each sample Insert a single nucleotide variant (FreeBayes quality > 20) into the reference genome. This was constructed using pyGeno. Sample-specific sequences of expressed proteins (tpm>0 ) was added to the canonical cancer proteome in fasta format.

[0084] To generate cancer-specific proteomes, trimmed R1 reads are processed using FASTX. -Use Toolkit version 0.0.14 to make it inversely complementary and trimmed R k-mer databases of 33-nucleotide and 24-nucleotide lengths with 2 reads. Used to generate the array. To exclude array determination errors and limit the database size. Therefore, the sample-specific threshold for the generation of the minimum k-mer is set as follows, relative to 33 nucleotides. Applied to: HGSC1~3: 7, OV642: 8, OV633: For 10, 4 for HGSC4 and OV606, 6 for HGSC5, HGS For C6, the answer is 5, and for OvCa48-114, the answer is 3. In human thymic epithelial cells (TECs) After subtracting the expressed k-mers to obtain cancer-specific k-mers, NEKTAR (in-house) The developed software can be found at https: / / github.com / iric-soft Longer arrays (continued) can be processed by the kmer_assembly tool ( / nektar) It was assembled into a 34-nucleotide contig, which was then translated into amino acids in 3 frames. The sequence was then split at the internal stop codon. The resulting chain of at least eight amino acid lengths was related. It was included in the cancer-specific proteome.

[0085] Isolation of MAP. Tumor and tissue samples are cut into small pieces (cubes, approximately 3 mm in size) and then tanned. Ice-cold protein inhibitor cocktail (Sigma, catalog number P8340-5ml) 5 ml of PBS was added. First, the Ultra Tu was set to a speed of 20000 rpm. Using rrax(trademark) T25 homogenizer (IKA-Labortechnik) Then for 20 seconds, then set the speed to 25,000 rpm and run the Ultra Turrax (commercial Using a standard T8 homogenizer (IKA-Labortechnik), for 20 seconds, 2 The mixture was homogenized multiple times. Then, 550 μl of ice-cold 10-fold lysis buffer (5% w / v CHAPS) was added. The following was added to each sample. After incubation at 4°C for 60 minutes while tumbling, The sample was centrifuged at 10,000 g at 4°C for 30 minutes. The supernatant was mixed with 1 mg of W6 / 32 antibody. Transfer the MAP to a new tube containing the covalently bound protein A magnetic beads, and then proceed as described above. Sea urchin (28) was immunoprecipitated. Then, the MAP extract was processed using Speed-Vac. They were dried and kept frozen before MS analysis.

[0086] MS analysis. Dried peptide extract was resuspended in 0.2% formic acid. , homemade C18 analytical column (C18 Jupiter Phenomenex(trademark)) A 15cm x 150μm inner diameter container filled with 0-30% acetonitrile (0.2% A 56-minute gradient from formic acid and 600 nl on the Easy-nLC II system. - 1 The sample was packed at the following flow rate. The sample was then measured using a Q-Exactive (trademark) HF mass spectrometer (Ther The data was analyzed using a Fisher Scientific instrument. The data was acquired at a resolution of 60,000. Each complete MS spectrum is followed by 20 MS / MS spectra, totaling 30,000 Resolution of 5 × 104 Automated gain control target, 100 ms injection time, and 25 % collision energy were selected for the most abundant polyvalent ions for MS / MS sequencing For HGSC4 - 6, after each full MS spectrum acquired at 60,000 resolution, 20 MS / MS spectra followed, with a resolution of 30,000, 2×10 Automated gain control target, 800 ms injection time, and 25% collision energy were selected for the most abundant polyvalent ions for MS / MS sequencing with 4 automation MAP identification. Peptides were identified using PEAKS 8.5 or Peaks X (Bioinformatics Solution Inc.), and the peptide sequences were searched against the global cancer database. For peptide identification, the tolerances for precursor ions and fragment ions were set to 10 ppm and 0.01 Da respectively. For samples from Schuster et al. (13), the tolerances for precursor ions and fragment ions were set to 5 ppm and 0.5 Da respectively. Oxidation (M) and deamidation ( NQ) occurrences were considered as post - translational modifications. To ensure that the MAP list contained only 5% of decoy identifications, a sample - specific threshold was applied to the PEAKS score. Peptides exceeding this threshold were further filtered according to the following criteria: peptide length 8 - 1

[0087] 1 amino acid, and MHC allele affinity rank based on NetMHC4.0 prediction ≦2% (29). TSA candidate identification and verification. To identify TSA candidates, each MAP and its corresponding coding For samples from Schuster et al., the tolerances for precursor ions and fragment ions were set to 5 ppm and 0.5 Da respectively. Oxidation (M) and deamidation ( NQ) occurrences were considered as post - translational modifications. To ensure that the MAP list contained only 5% of decoy identifications, a sample - specific threshold was applied to the PEAKS score. Peptides exceeding this threshold were further filtered according to the following criteria: peptide length 8 - 1 1 amino acid, and MHC allele affinity rank based on NetMHC4.0 prediction ≦2% (29). 1 amino acid, and MHC allele affinity rank based on NetMHC4.0 prediction ≦2% (29). ≦2% (29).

[0088] TSA candidate identification and verification. To identify TSA candidates, each MAP and its coding The sequences are, respectively, associated with cancer and normal canonical proteome, or cancer The database of normal 24-nucleotide k-mers was also queried. d) Typical canonical proteome and typical 24-nucleotide k-mer data. The tabes were derived from RNA-Se24, obtained from purified TEC extracted from six human thymes. Constructed using the lead of q (15 and U.S. Provisional Application No. 62 / 724,760). MA P is in two cases: i) in the normal canonical proteome of the sample, and also in the normal (i.e.) In cases where the peptide sequence was not detected even in the TEC)k-mer, or i i) The peptide is not present in both cancer and normal canonical proteomes, and RNA coding sequences are overexpressed at least 10 times more in cancer cells compared to TECs. If so, it was labeled as a TSA candidate. MAP corresponds to several RNA sequences. In that case, it is only if all sequences match the TSA candidate status. It was considered a candidate for SA. The MS / MS spectra of all TSA candidates were manually verified. Any false identifications were removed. Supported by RNA data that could be distinguished by MS. Among the TSA candidates possessing the I / L mutant, the most expressed mutant is the TSA candidate. If so, both mutants were further examined.

[0089] Finally, mapping reads containing MAP coding sequences on the reference genome (GRCh38) By mapping using BLAT (UCSC Genome Browser), the genome position The placement was assigned to all MS-validated TSA candidates. Hypervariable region (HLA, Ig or T) TSA candidates for reads that matched the CR gene were excluded. TSA candidates are MAP code The sequence contains mutants that do not match known germline polymorphisms (reported in dbSNP v149). If present, it was classified as mTSA. Non-mutational candidates were classified as aeTSA candidates, and correct This allowed for further evaluation of their expression in normal tissues and organs.

[0090] Tissue expression of sequences encoding aeTSA candidates. RNA-Seq data from 27 different tissues. The data is available through the Genotype-Tissue expression (GTEx) portal. Downloaded from (phs000424.v7 accessed on April 16, 2018) .p2), as mentioned above (15), in order to evaluate the expression of the coding sequence of aeTSA candidate The RNA-Seq data was collected from the cervix (n=6), fallopian tubes (n=7), and adipose tissue. Excluding the bladder (n=49), bladder (n=12), and kidneys (n=38), from 50 donors Obtained. The registration numbers of the GTEx datasets used in this study are listed in Table 2. Briefly described below. Therefore, the number of reads that completely cover the MAP coding sequence is the RNA-Seq reads of each tissue. 24-me MAP code sequence in a database of 24-mer transformed from code The read count was estimated by the minimum occurrence of r sets. Read counts were calculated per 100 million sequences determined. Normalize to the read (rphm), and then logarithmically transform (log 10 (rphm+1), This was averaged across all available RNA-Seq experiments for each tissue. MHC 低 Peripheral expression of rphm > 10 was observed in tissues other than the cerebral cortex, nerves, and testes. We considered any aeTSA candidate that did not exist to be a genuine aeTSA. [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4]

[0091] Expression of the TSA coding region. RNA expression of the TSA coding region on the TSA coding strand. As a count of all reads overlapping with the A chord area, parameter orientation = "same" The mapping is done using the `qCount` function of the R package `QuasR` (30). The data was quantified from the BAM files. The count was calculated per 100 million mapped reads. The number of reads was normalized. In aeTSA expression analysis, the context of surrounding sequences is considered. Since t) can affect aeTSA expression, individual aeTSA molecules are mapped to We specifically analyzed the NEEC domain.

[0092] Clinical and genomic data from TCGA. hg38 for HM27 methylation, DNA Copy number variation, RNA-Seq gene expression, and clinical data were processed and normalized. The converted Level 3 data was processed using the R package 'TCGAbiolinks' (31) I downloaded it from the TCGA Open Access Database. (Arm-level DNA copy) The number of changes was analyzed by the Broad Institute TCGA Genome Data Analysis Center (do Downloaded from i:10.7908 / C1P84B9Q.

[0093] Immune cell score. An immune cell score representing the immune cell population is calculated based on RNA-Seq data. This was estimated. For each tumor, the score was explained by Danaher et al. (32). Therefore, it was estimated as the average of the logarithmically transformed FPKM values ​​of those marker genes.

[0094] Frequency of aeTSA presentation. To estimate the number of aeTSA presentations by individual patients. Bioinformatics simulations were then performed. This estimation uses two parameters. It was based on the following. Firstly, the possibility of aeTSA expression was in the TCGA-OV cohort. It was based on the proportion of tumors expressing the corresponding RNA. a The expression likelihood for eTSA was calculated as follows: (TCGA expressing aeTSA) (Percentage of tumors) × (SNP frequency in a given population) SNP frequency is calculated using Genome Aggregation. Obtained from the Region Database. Secondly, a collection of European Americans. Group (n=1,242,890), African Americans (n=416,581), and Chinese ( For n=99672, HLA allele frequencies were obtained from the National Marrow Bank (NMDP). We got it. Next, we determined the patient's HLA genotype based on the reported frequency in a given population. This was simulated with six HLA class I alleles. The six HLA alleles were independently Because it was assumed that this was the case, some HLA loci were homozygous in the simulated patients. The condition was conjugated. When both aeTSA and the associated HLA alleles were expressed, ae TSA was considered to be presented to the simulated patient. Expression of each aeTSA was considered to be in the same overlapping group. Except for duplicate aeTSA, whose expression state was simulated only once, these are independent events. This was considered to be the case. 1 million simulated patients and their aeTSA presentation status were divided into three groups. It was generated and used to plot the distribution.

[0095] Statistical analysis and data visualization. Analysis and numerical data are performed using R v3.5.1 or Python. This was done using n v2.7.6. The 'gplots' package in R was used to plot the tumor A heatmap of TSA coding region expression in ulcers was generated. Correlation studies were performed as instructed. Unless otherwise specified, the method was performed using Spearman's method and the R function 'cor.test'. The ANOVA test was used for the comparison of fabrics, and pairwise comparisons between groups were performed using the Ulcoxon method. The test was performed using a rank-sum test. The log-rank p-values ​​for survival analysis were calculated using the 'survival' package. I calculated it using a cage.

[0096] Example 2: Proteogenetic analysis identified 111 TSAs in 23 HGSCs. To obtain a system-level characterization of the TSA landscape, high-throughput tank Direct MAP identification was performed using Dem MS (MS / MS) analysis (34-36). The search engine matches each acquired MS / MS spectrum to the peptide sequence. , depends on the user-defined protein database (37). Therefore, in the test sample The peptide is only searchable by search engines if its sequence is included in the reference database. It can be identified. General reference protein databases such as UniProt can identify the sample. Because it does not contain specific mutations, out-of-frame translation events, and non-exon sequences, all G The search for capturing TSA encoded by the nom region is tumor-specific for each tumor. This requires building a customized database containing different translation products. Each sample analyzed using the recently described proteogenetic approach (15) A customized search database was constructed from RNA-Seq readings. The customized database consists of two modules: canonical proteome. It contains (intraframe translation of exons) and cancer-specific proteome, which is (TEC This is a 3-frame translation of the cancer-specific RNA sequence after subtracting the normal RNA sequence (from) (Figure). 1) TECs are important in i) establishing immune tolerance during the development of immature T cells. ii) its role (i.e., tolerance to medium water), and ii) more transcripts than other types of somatic cells. It was used as a normal control for two reasons: its remarkable ability to express promiscuously (38). After obtaining MAPs from nine primary HGSC samples by immunoprecipitation of MHC I molecules, Analysis was performed by liquid chromatography-MS / MS (15, 28). Immunopeptemia in an additional cohort of 14 HGSCs reported by r et al. (13) The data also includes matching RNA-Seq and MS data available for samples as described herein. Each TSA candidate was re-analyzed by applying a proteogenomic approach. These were confirmed by manual validation of the spectrum and genome location.

[0097] Candidate duplicate genomic variants not present in dbSNPs may represent germline polymorphisms. Low, and therefore labeled as mTSA. This classification was used for the 23 analyses. Eighteen mTSAs were obtained from the sample (Table 3A). All of these mTSAs have been previously reported. Not done. Of the 18 mTSAs, 7 originate from intraframe exon translation, and 4 Some originated from out-of-frame exon translation, and eight originated from non-coding sequences. Regarding the three tumors... Therefore, if a consistent normal tissue is available, RNA-Seq analysis will show that the mTSA variant is reproductively affected. It was confirmed that it was not a cell lineage polymorphism (Figures 8A-8B). Tumors in which normal tissue does not match In some cases, some mTSAs may correspond to rare polymorphisms that are not present in dbSNPs. This cannot be formally ruled out. The number of mTSAs may be slightly overestimated. In cases where there is less than one mTSA per tumor (18 mTSAs / 23 tumors), mTS The conclusion is that A is rare in HGSC and therefore represents a less attractive target. It only makes it stronger. Furthermore, the classic TSA detection method is frame exon translation or Because we are strictly focusing on mTSAs resulting from these, they are reported herein. It will only reveal 7 out of 111 TSAs. [Table 3]

[0098] For non-mutant TSA candidates, strict criteria are applied to determine pure aeTSA, i.e., They identified cancer-specific expression of [specific gene]. Following this, they identified 27 cells throughout the human body. We analyzed the expression of these RNAs in peripheral tissues. MHC 低 Other than tissues (brain, nerves, testes) All candidate candidates (rphm>10) whose coding RNA is expressed in any peripheral tissue are selected. Therefore, it was removed from the aeTSA list. The coding sequence of the aeTSA candidate was germline single nucleotide polymorphism. If it contains the type (as reported in dbSNP), this candidate is an SNP-containing sequence. And only if the reference sequence meets the above criteria, it is labeled as a valid aeTSA. (Figure 9). Overall, 93 aeTSA candidates met these stringent criteria (Figure 2). (Tables 3B and 3C), of which 85 have never been reported to our knowledge. Table 2B). Interestingly, of the 93 aeTSAs, 5 were expressed in the testes. While several cancer bacterial antigens (CGAs) are aeTSAs, most aeTSAs are CGAs It was shown that this is not the case. CGA is a canonical expression normally expressed only by germ cells. Encoded by Son, their abnormal expression in cancer cells is primarily epigenetic. It is promoted by certain changes. However, some CGAs are expressed by adult mTECs. (16) CGA expressed in mTEC (or other body tissue) is expressed as TAA. Anything not expressed by any normal tissue (including mTECs) is a genuine aeTS. It is considered to be A.

[0099] A genomic location was assigned to each aeTSA. When multiple locations were possible, matching RNA was used. The one with the highest read generation rate was selected. All TSA characteristics are shown in Tables 3B and 3C. Report to: The stringent approach is atypical translation (5'UTR, 3'UTR, The total number of aeTSAs resulting from intergenetic and frameshifts is formally underestimated. It is possible. In fact, the read frames used to generate MAPs in tumors are As is known, when these coding RNAs are expressed in some normal tissues, It is not possible to guess whether the reading frame can be translated. Therefore, TS To avoid including false positives in list A, such aeTSA candidates were excluded. . [Table 4-1] [Table 4-2] 1 The "translation event" column summarizes the relationship between the TSA code sequence and the ORF, with the outside of the ORF being "non-coding", the overlapping but frame-shifted ORF being "code out", and the ORF-matched being "code in". atching being "code in". 2 The "genomic origin" column further annotates the aeTSA according to the biotype from Ensembl. For example, "antisense" means that the sequence is on the opposite strand as the annotated gene, "canonical" refers to the canonical / annotated ORF, and "ncRNA" is an annotated RNA that does not contain an ORF according to Ensembl. strand, "canonical" refers to the canonical / annotated ORF and "ncRNA" is an annotated R NA that does not contain an ORF according to Ensembl. 3 "ncRNA" refers to the case where the TSA code sequence aligns with the exon of the non-coding transcript. 4 "Non-coding antisense" means that the TSA code sequence is the antisense of the gene in the Ensembl annotation, and thus it is a non-coding region.

Table 5

[0100] Example 3: Most of the TSA of HGSC are non-mutated MAPs obtained from non-canonical translation. A total of 111 unique TSAs were identified by an average of 2,200 unique MAPs identified per sample (Figure 3A). The number of TSAs identified per sample was significantly correlated with the number of MAPs (Figure 3B). Furthermore, there was a moderate correlation between the number of MAPs per HLA allele and the tumor sample size (Figure 10). This indicates that the tumor sample size has an impact on MS analysis. and the number of MAPs (Figure 3B). Furthermore, there was a moderate correlation between the number of MAPs per HLA allele and the tumor sample size ​​​​​​​This is consistent with the idea that it is a limiting factor (28). In principle, mutated non-coding A TSA derived from a column can be specified as either an mTSA or an aeTSA. We decided to label them as mTSAs as appropriate. The basis for this is their genome origin ( Regardless of whether it is exonential or not, mTSA is a "private TSA". In other words, it was expected that it would not be shared by multiple tumors. In contrast, The variant aeTSA could theoretically be shared by a significant proportion of HGSCs.

[0101] Notably, the first to be processed in this study, or by Schuster et al. The TSA characteristics identified in the treated samples were remarkably similar (Figure 3C). This is because the proteogenetic approaches described herein are generally RNA-Seq and This suggests that it can be applied to MS data and is superficially unaffected by inter-laboratory variability. This suggests that in both cohorts, approximately 83% of TSAs were not mutated, and the majority of TSAs The minutes arose from atypical translation: mainly from non-code areas, and in a small part, from fre It was derived from an exotranslation of the system (Figure 3C). Two features of aeTSA are noteworthy: i) 80% are non-coding sequences, particularly introns (31%) and intergenes (22%). ii) 90% are new MAPs (Figure 3D). Previously reported MAPs are compared to The corresponding protein isoforms are found in the UniProt database (13, 39-43). Processed transfer (E) included in U.S. Patent Publication No. 2012 / 0077696A1 Except for those matching the biotype annotated by the nsembl database, Derived from exon translation within the frame.

[0102] Example 4: Expression of aeTSA-coding transcripts in ovarian cancer samples. Does cancer-specific expression of aeTSA-coding transcripts arise from random transcription noise? To determine whether this is due to repetitive transcriptional abnormalities, this study and TCGA eggs were used to determine whether it is caused by repetitive transcriptional abnormalities. Genomic regions encoding 93 aeTSAs identified in samples from the tumor cohort. RNA expression was analyzed. The region encoding aeTSA was found in a significant proportion of ovarian cancers. It was expressed in at least 10% of the samples, with 72 cells (77%) being expressed. Sixteen cells (17%) were expressed in 80% of cases (Figure 4). These commonly expressed regions are located in the affected area. It is highly likely that shared TSAs will be generated between parties. Therefore, 93 ae in HGSC The expression of this set of TSA-coding transcripts is not a rare or random event, but rather a significant event. It can be concluded that this is a common characteristic of HGSC.

[0103] Example 5: Genomic correlation of aeTSA expression. To understand the mechanism of aeTSA expression, we used the TCGA-OV dataset. Using luciomic data, aeTSA RNA expression and local gene or epigei We explored the relationship with genetic abnormalities. Where applicable, local DNA copy number changes. DNA methylation levels on gene promoter regions, and RNA for each aeTSA The correlation between expression levels was tested (Figure 5A). Genomic regions (exons, genes) that are part of a gene were examined. When using aeTSA derived from TRON (or UTR), the expression of related genes and We also analyzed the correlation between the gene and aeTSA expression. In the latter situation, the relationship between the gene and aeTSA expression and A significant correlation was observed between them (Figure 5A). This is because the coding region is located within the gene. For SA, the regulation of aeTSA expression generally suggests that it affects the entire gene. Furthermore, changes in DNA copy number showed a positive correlation with the RNA expression level of aeTSA. This is because intragenetic aeTSA and extragenetic aeTSA (antisense and gene The same was true in both cases. This indicates that changes in DNA copy number have a substantial impact on aeTSA expression. This suggests an influence. In particular, this correlation is expressed in a larger proportion of tumors. The staining was particularly strong for intragenetic aeTSA (Figure 11A). Upon examining the chromosome distribution, it was found that several chromosome arms that are frequently amplified in HGSC are associated with many AEs. It was found that TSA was produced (Figure 5B). For example, it is commonly amplified in ovarian cancer. The long arm (44) of chromosome 3 was the source of eight aeTSAs. One example is MECOM, located at 3q26.2(44), which has three overlapping exons. Out-of-frame aeTSA was generated (Table 3B). However, amplification of chromosome arms is not always possible. It was not necessary (for example, 15q), and it was not enough to generate aeTSA (for example) (8q) (Figure 5B, Figure 11B).

[0104] For the technology that TCGA uses for DNA methylation (HM27 array) analysis, genes For the promoters of external aeTSA and some aeTSA source genes, Promoter methylation data was unavailable. Therefore, the promoter methylation analysis was performed on 17 individuals. This was limited to a subset of aeTSA. Nevertheless, for six aeTSA, DNA analysis was performed. A significant correlation was found between chilling and aeTSA expression (Figure 5A). The correlation was observed in 5 cases. The results were negative, with one case being positive. This indicates that promoter demethylation frequently enhances transcription. This aligns with the idea that it brings about. In particular, the two genes showing the highest negative correlation are MAGEC1(ρ=-0.53, P adj = 1.6 x 10 -26 ) and MAGEA4( ρ = -0.51, P adj = 6.7 x 10 -25 These are accompanied by the arrows in Figure 5A. It is represented by a dark bar. The MAGE family of genes is found in several cancers, including HGSC. It is a CGA that is overexpressed in the ip (3). Overall, aeTSA expression is at least Partially, this is regulated at the transcriptional level by variations in gene copy number and DNA methylation. We can conclude that this is the case.

[0105] Example 6: Expression of three aeTSA molecules correlates with improved survival rates. Next, we evaluated whether several aeTSAs could trigger a spontaneous protective immune response. aeTSA expression at the cytoplasmic level, in addition to aeTSARNA expression, is associated with HL The fact that the presence of the A allotype is required makes addressing this problem complicated. Therefore, patients from the TCGA cohort were selected based on the expression of individual aeTSA RNAs. Based on (or not based on) and the presence of (or based on) the associated HLA allotype. It was subdivided into four subgroups. The presentation of three aeTSAs is more preferable. It correlated with clinical outcomes (Figures 6A-6C). HLA allele polymorphisms affected the size of each group. This reduced the power of the analysis, thus decreasing its detection power. The log-rank p-value for SA was in the range of 0.013 to 0.076 (Figures 6A to 6A). 6C). Nevertheless, the two observations suggest that these correlations are not biologically significant. This provides evidence to support the claim. Firstly, the "protective effect" of these aeTSAs is HL It appeared to be restricted to A: In patients expressing aeTSA RNA, the associated H When the LA allele was also expressed, the survival rate was superior. Secondly, RTHQMNTFQR Expression of aeTSA and its associated HLA allotypes is associated with T cells and cytotoxic T cells. This showed a positive correlation with tumor invasion by cytoplasm (Figure 6D, Figure 6E); ANOVA, p<0.05). .

[0106] Example 7: Median number of aeTSAs presented by individual tumors Using the list of 93 aeTSA, we ultimately determined how this study relates to TSA-targeted immunotherapy. We estimated the extent of the potential benefits. Therefore, 93% of 1 million patients The presentation states of individual aeTSA were randomly simulated. The frequencies of HLA alleles were estimated. To determine this, we need three of the largest datasets from the US bone marrow bank: European American. Humans, African Americans, and Chinese (45) were used. Alleles in a given population Frequency, expression rate in TCGA-OV tumors, and, where applicable, SNP frequency are used. Then, six HLA alleles and aeTSA expression states were independently generated for each tumor. The number of aeTSA molecules per cell was calculated as the sum of the expressed HLA-aeTSA pairs. Based on these simulations, 98% of European whites and African Americans It was determined that at least one aeTSA could be found in 74% and 78% of Chinese individuals. Furthermore, the median number of aeTSAs per tumor is 5 for European Caucasians and 5 for African Americans. The differences were 2 in humans and 4 in Chinese (Figure 7). These differences between groups were due to HLA alleles. Changes in the frequency of offspring, and the fact that tumor samples are primarily from European Caucasians This occurred. These calculations underestimated the number of aeTSAs per tumor, mainly for three reasons. There is suspicion that this is the case. Firstly, more than 50% of MAPs bind to two or more HLA allotypes. However, in many cases, they bind across supertypes or loci (46) However, this is because it has not been taken into consideration. Secondly, the genomic region encoding a given MAP is This is because it frequently generates duplicate MAPs presented by different HLA allotypes (23) Yes. Thirdly, regarding the five aeTSAs, including non-synonymous SNPs listed in dbSNP: Furthermore, only SNP mutants that generate MAPs in the sample are effective, and other SNP mutants are M We assumed that AP was not produced. A single amino acid change invalidates MAP presentation. This cautious strategy was adopted because it was likely sufficient (47). The current 93 a Vaccines containing the eTSA set are available to almost all whites and Africans who have HGSC. It was concluded that this covers a significant proportion of both Caucasian Americans and Asians (e.g., Chinese). It is possible to do so.

[0107] The technology of the present invention is described above by specific embodiments thereof, but the appended claims The subject invention may be modified without departing from the spirit and nature of the subject invention as defined within the scope of the subject invention. In patent claims, the word "comprising" means "including but , but not limited to The expression is essentially equivalent to "open-ended," and is used as a term for open-ended systems. It is used. The singular forms "a," "an," and "the" are used when the context makes it clear that they are not singular. Unless otherwise specified, it includes multiple corresponding references.

[0108] References 1.Jayson GC,Kohn EC,Kitchener HC,Lederma nn JA.Ovarian cancer.Lancet 2014;384:137 6-88 2.Bowtell DD, Bohm S, Ahmed AA, Aspuria PJ, Bast RC,Jr.,Beral V,et al.Rethinking ova rian cancer II:reducing mortality from h igh-grade serous ovarian cancer.Nat Rev Cancer 2015;15:668-79 3.Want MY, Lugade AA, Battaglia S, Odunsi K .Nature of tumor rejection antigens in ovarian cancer.Immunology 2018;155:202-1 0 4. Yang SYC, Lheureux S, Karakasis K, Burnie r JV,Bruce JP,Clouthier DL,et al.Landsca pe of genomic alterations in high-grade serous ovarian cancer from exceptional l on-and short-term survivors.Genome Med 2018;10:81 5. Zhang AW, McPherson A, Milne K, Kroeger D R,Hamilton PT,Miranda A,et al.Interfaces of Malignant and Immunologic Clonal Dyn amics in Ovarian Cancer.Cell 2018;173:17 55-69 e22 6.Hamanishi J,Mandai M,Ikeda T,Minami M, Kawaguchi A,Murayama T,et al.Safety and Antitumor Activity of Anti-PD-1 Antibody ,Nivolumab,in Patients With Platinum-Res istant Ovarian Cancer.J Clin Oncol 2015; 33:4015-22 7.Hamanishi J,Mandai M,Konishi I.Immune checkpoint inhibition in ovarian cancer. Int Immunol 2016;28:339-48 8.Rodriguez-Garcia A,Minutolo NG,Robinso n JM,Powell DJ.T-cell target antigens ac ross major gynecologic cancers.Gynecol O ncol 2017;145:426-35 9.Riley RS,June CH,Langer R,Mitchell MJ. Delivery technologies for cancer immunot herapy.Nat Rev Drug Discov 2019 10.Ehx GE,Perreault C.Discovery and char acterization of actionable tumor antigen s.Genome Med 2019;11:1-3 11.Millar DG,Ohashi PS.Central tolerance :what you see is what you don’t get! Nat Immunol 2016;17:115-6 12.Haen SP,Rammensee HG.The repertoire o f human tumor-associated epitopes--ident ification and selection of antigens and their application in clinical trials.Cur r Opin Immunol 2013;25:277-83 13.Schuster H,Peper JK,Bosmuller HC,Rohl e K,Backert L,Bilich T,et al.The immunop eptidomic landscape of ovarian carcinoma s.Proc Natl Acad Sci U S A 2017;114:E994 2-E51 14.Laumont CM,Daouda T,Laverdure JP,Bonn eil E,Caron-Lizotte O,Hardy MP,et al.Glo bal proteogenomic analysis of human MHC class I-associated peptides derived from non-canonical reading frames.Nat Commun 2016;7:10238 15.Laumont CM,Vincent K,Hesnard L,Audema rd E,Bonneil E,Laverdure JP,et al.Non-co ding regions are the main source of targ etable tumor-specific antigens.Sci Trans l Med 2018;10:aau5516 16.Gotter J,Brors B,Hergenhahn M,Kyewski B.Medullary epithelial cells of the hum an thymus express a highly diverse selec tion of tissue-specific genes colocalize d in chromosomal clusters.J Exp Med 2004 ;199:155-66 17.Bobisse S,Genolet R,Roberti A,Tanyi J L,Racle J,Stevenson BJ,et al.Sensitive a nd frequent identification of high avidi ty neo-epitope specific CD8(+)T cells in immunotherapy-naive ovarian cancer.Nat Commun 2018;9:1092 18.Simoni Y,Becht E,Fehlings M,Loh CY,Ko o SL,Teng KWW,et al.Bystander CD8(+)T ce lls are abundant and phenotypically dist inct in human tumour infiltrates.Nature 2018;557:575-9 19.Marty R,Kaabinejadian S,Rossell D,Sli fker MJ,van de Haar J,Engin HB,et al.MHC -I Genotype Restricts the Oncogenic Muta tional Landscape.Cell 2017 20.Capietto AH,Jhunjhunwala S,Delamarre L.Characterizing neoantigens for persona lized cancer immunotherapy.Curr Opin Imm unol 2017;46:58-65 21.Bilich T,Nelde A,Bichmann L,Roerden M ,Salih HR,Kowalewski DJ,et al.The HLA li gandome landscape of chronic myeloid leu kemia delineates novel T-cell epitopes f or immunotherapy.Blood 2019;133:550-65 22.Loffler MW,Mohr C,Bichmann L,Freudenm ann LK,Walzer M,Schroeder CM,et al.Multi -omics discovery of exome-derived neoant igens in hepatocellular carcinoma.Genome Med 2019;11:1-16 23.Pearson H,Daouda T,Granados DP,Durett e C,Bonneil E,Courcelles M,et al.MHC cla ss I-associated peptides derive from sel ective regions of the human genome.J Cli n Invest 2016;126:4690-701 24.Szolek A,Schubert B,Mohr C,Sturm M,Fe ldhahn M,Kohlbacher O.OptiType:precision HLA typing from next-generation sequenc ing data.Bioinformatics 2014;30:3310-6 25.Bolger AM,Lohse M,Usadel B.Trimmomati c:a flexible trimmer for Illumina sequen ce data.Bioinformatics 2014;30:2114-20 26.Garrison E,Marth G.Haplotype-based va riant detection from short-read sequenci ng.arXiv 2012;1207.3907[q-bio.GN] 27.Daouda T,Perreault C,Lemieux S.pyGeno :A Python package for precision medicine and proteogenomics.F1000Res 2016;5:381 28.Lanoix J,Durette C,Courcelles M,Cosse tte E,Comtois-Marotte S,Hardy MP,et al.C omparison of the MHC I immunopeptidome r epertoir of B-cell lymphoblasts using tw o isolation methods.Proteomics 2018;18:e 1700251 29.Andreatta M,Nielsen M.Gapped sequence alignment using artificial neural netwo rks:application to the MHC class I syste m.Bioinformatics 2016;32:511-7 30.Gaidatzis D,Lerch A,Hahne F,Stadler M B.QuasR:quantification and annotation of short reads in R.Bioinformatics 2015;31 :1130-2 31.Colaprico A,Silva TC,Olsen C,Garofano L,Cava C,Garolini D,et al.TCGAbiolinks: an R / Bioconductor package for integrativ e analysis of TCGA data.Nucleic Acids Re s 2016;44:e71 32.Danaher P,Warren S,Dennis L,D’Amico L ,White A,Disis ML,et al.Gene expression markers of Tumor Infiltrating Leukocytes .J Immunother Cancer 2017;5:18 33.Perez-Riverol Y,Csordas A,Bai J,Berna l-Llinares M,Hewapathirana S,Kundu DJ,et al.The PRIDE database and related tools and resources in 2019:improving support for quantification data.Nucleic Acids R es 2019;47:D442-d50 34.Shao W,Pedrioli PGA,Wolski W,Scurtesc u C,Schmid E,Vizcaino JA,et al.The Syste MHC Atlas project.Nucleic Acids Res 2018 ;46:D1237-D47 35.Gfeller D,Bassani-Sternberg M.Predict ing Antigen Presentation-What Could We L earn From a Million Peptides? Front Immu nol 2018;9:1716 36.Villani AC,Sarkizova S,Hacohen N.Syst ems Immunology:Learning the Rules of the Immune System.Annu Rev Immunol 2018;36: 813-42 37.Caron E,Kowalewski DJ,Chiek Koh C,Stu rm T,Schuster H,Aebersold R.Analysis of Major Histocompatibility Complex(MHC)Imm unopeptidomes Using Mass Spectrometry.Mo l Cell Proteomics 2015;14:3105-17 38.Sansom SN,Shikama-Dorn N,Zhanybekova S,Nusspaumer G,Macaulay IC,Deadman ME,et al.Population and single-cell genomics reveal the Aire dependency,relief from P olycomb silencing,and distribution of se lf-antigen expression in thymic epitheli a.Genome Res 2014;24:1918-31 39.Bassani-Sternberg M,Pletscher-Frankil d S,Jensen LJ,Mann M.Mass spectrometry o f human leukocyte antigen class I peptid omes reveals strong effects of protein a bundance and turnover on antigen present ation.Mol Cell Proteomics 2015;14:658-73 40.Bassani-Sternberg M,Braunlein E,Klar R,Engleitner T,Sinitcyn P,Audehm S,et al .Direct identification of clinically rel evant neoepitopes presented on native hu man melanoma tissue by mass spectrometry .Nat Commun 2016;7:13404 41.Rizvi NA,Hellmann MD,Snyder A,Kvistbo rg P,Makarov V,Havel JJ,et al.Cancer imm unology.Mutational landscape determines sensitivity to PD-1 blockade in non-smal l cell lung cancer.Science 2015;348:124- 8 42.Gloger A,Ritz D,Fugmann T,Neri D.Mass spectrometric analysis of the HLA class I peptidome of melanoma cell lines as a promising tool for the identification o f putative tumor-associated HLA epitopes .Cancer Immunol Immunother 2016;65:1377- 93 43.UniProt Consortium:a worldwide hub of protein knowledge.Nucleic Acids Res 201 9;47:D506-D15 44.Cancer Genome Atlas Research Network. Integrated genomic analyses of ovarian c arcinoma.Nature 2011;474:609-15 45.Maiers M,Gragert L,Klitz W.High-resol ution HLA alleles and haplotypes in the United States population.Hum Immunol 200 7;68:779-88 46.Rao X,Hoof I,Costa AI,van Baarle D,Ke smir C.HLA class I allele promiscuity re visited.Immunogenetics 2011;63:691-701 47.Granados DP,Sriranganadane D,Daouda T ,Zieger A,Laumont CM,Caron-Lizotte O,et al.Impact of genomic polymorphisms on th e repertoire of human MHC class I-associ ated peptides.Nat Commun 2014;5:3600 48.Delaney JR,Patel CB,Willis KM,Haghigh iabyaneh M,Axelrod J,Tancioni I,et al.Ha ploinsufficiency networks identify targe table patterns of allelic deficiency in low mutation ovarian cancer.Nat Commun 2 017;8:14423 49.Kahles A,Lehmann KV,Toussaint NC,Huse r M,Stark SG,Sachsenberg T,et al.Compreh ensive Analysis of Alternative Splicing Across Tumors from 8,705 Patients.Cancer Cell 2018;34:211-24 e6 50.Ali M,Foldvari Z,Giannakopoulou E,Bos chen ML,Stronen E,Yang W,et al.Induction of neoantigen-reactive T cells from hea lthy donors.Nat Protoc 2019 51.Croft NP,Smith SA,Pickering J,Sidney J,Peters B,Faridi P,et al.Most viral pep tides displayed by class I MHC on infect ed cells are immunogenic.Proc Natl Acad Sci U S A 2019;116:3112-7.

Claims

1. A tumor antigen peptide containing one of the amino acid sequences described in SEQ ID NOs: 1 to 103.

2. The tumor according to claim 1, comprising one of the amino acid sequences described in SEQ ID NOs: 19 to 103. Cerebral antigen peptide.

3. The tumor antigen peptide binds to the HLA-A*01:01 molecule, as shown in SEQ ID NOs. 21 and 28. The amino acid sequence according to claim 1 or 2, comprising the amino acid sequence described in 40, 41, 66, or 88. Tumor antigen peptide.

4. The tumor antigen peptide binds to the HLA-A*02:01 molecule, and SEQ ID NOs: 1, 19, 20, 22, 30, 31, 36, 50, 52, 60, 62, 73, 84, 85, 86 also The tumor antigen peptide according to claim 1 or 2, comprising the amino acid sequence described in 91.

5. The tumor antigen peptide binds to the HLA-A*11:01 molecule, as shown in SEQ ID NOs. 32 and 54. Claims including the amino acid sequence described in 55, 67, 69, 81, 87, 90, or 102. The tumor antigen peptide described in item 1 or 2.

6. The tumor antigen peptide binds to the HLA-A*24:02 molecule, and SEQ ID NO: 33 or The tumor antigen peptide according to claim 1 or 2, comprising the amino acid sequence described in 43.

7. The tumor antigen peptide is bound to the HLA-A*25:01 molecule, as described in Sequence ID No.

24. The tumor antigen peptide according to claim 1 or 2, comprising one of the amino acid sequences of the following.

8. The tumor antigen peptide binds to the HLA-A*29:02 molecule, and SEQ ID NO: 34 or The tumor antigen peptide according to claim 1 or 2, comprising one of the amino acid sequences described in 58. Chido.

9. The tumor antigen peptide is bound to the HLA-A*32:01 molecule, as described in Sequence ID No.

16. The tumor antigen peptide according to claim 1 or 2, comprising the amino acid sequence.

10. The tumor antigen peptide binds to the HLA-B*07:02 molecule, as described in Sequence IDs 4, 6, and 8. Claims including the amino acid sequence described in 9, 26, 49, 78, 92, 97, or 101. The tumor antigen peptide described in item 1 or 2.

11. The tumor antigen peptide binds to the HLA-B*08:01 molecule, as shown in Sequence IDs 23 and 35. The amino acids listed in 42, 44, 46, 59, 63, 70, 74, 76, 83, or 103 The tumor antigen peptide according to claim 1 or 2, comprising one of the sequences.

12. The tumor antigen peptide is bound to the HLA-B*14:01 molecule, as described in Sequence ID No.

53. The tumor antigen peptide according to claim 1 or 2, comprising the amino acid sequence.

13. The tumor antigen peptide binds to the HLA-B*15:01 molecule, and SEQ ID NOs: 2, 3 The tumor antigen peptide according to claim 1 or 2, comprising the amino acid sequence described in 5.

14. The tumor antigen peptide is bound to the HLA-B*18:01 molecule, as described in Sequence ID No.

89. The tumor antigen peptide according to claim 1 or 2, comprising the amino acid sequence.

15. The tumor antigen peptide binds to the HLA-B*39:01 molecule, as shown in SEQ ID NOs. 47 and 64. , comprising the amino acid sequence described in 96 or 99, the tumor antigen peptide according to claim 1 or 2 Chido.

16. The tumor antigen peptide binds to the HLA-B*40:01 molecule, as shown in SEQ ID NOs: 11, 12 Or the tumor antigen peptide according to claim 1 or 2, comprising the amino acid sequence described in 13.

17. The tumor antigen peptide is bound to the HLA-B*44:02 molecule, as described in Sequence ID No.

65. The tumor antigen peptide according to claim 1 or 2, comprising the amino acid sequence.

18. The tumor antigen peptide binds to the HLA-B*44:03 molecule, and SEQ ID NO: 37 or A tumor antigen peptide according to claim 1 or 2, comprising the amino acid sequence described in 94.

19. The tumor antigen peptide binds to the HLA-C*03:03 molecule, as shown in SEQ ID NOs: 10 and 29. , comprising the amino acid sequence described in 71 or 95, the tumor antigen peptide according to claim 1 or 2 Chido.

20. The tumor antigen peptide binds to the HLA-C*04:01 molecule, and SEQ ID NO: 6 or 1 A tumor antigen peptide according to claim 1 or 2, comprising the amino acid sequence described in 5.

21. The tumor antigen peptide is bound to the HLA-C*05:01 molecule, as described in Sequence ID No.

27. The tumor antigen peptide according to claim 1 or 2, comprising the amino acid sequence.

22. The tumor antigen peptide binds to the HLA-C*06:02 molecule, and SEQ ID NO: 18 or A tumor antigen peptide according to claim 1 or 2, comprising the amino acid sequence described in 72.

23. The tumor antigen peptide binds to the HLA-C*07:01 molecule, as shown in SEQ ID NOs: 38, 61. Or the tumor antigen peptide according to claim 1 or 2, comprising the amino acid sequence described in 93.

24. The tumor antigen peptide is bound to the HLA-C*07:02 molecule, as described in Sequence ID No.

7. A tumor antigen peptide according to claim 1 or 2, comprising an amino acid sequence.

25. The tumor antigen peptide is bound to the HLA-C*12:03 molecule, as described in Sequence ID No.

80. The tumor antigen peptide according to claim 1 or 2, comprising the amino acid sequence.

26. The tumor antigen peptide binds to the HLA-C*14:02 molecule, as shown in SEQ ID NOs. 25 and 57. Or the tumor antigen peptide according to claim 1 or 2, comprising the amino acid sequence described in 79.

27. Encoded by sequences located in the non-protein coding region of the genome, SEQ ID NOs: 1, 4 , 6-8, 10, 13, 15-27 and 36-99, preferably sequence numbers 19-27 Claims 1 to 26, comprising the amino acid sequence described in any one of 36 to 99. The tumor antigen peptide described in item 1.

28. The non-protein coding region of the genome is an untranslated transcription region (UTR), The tumor antigen peptide is preferably sequence numbers 1, 8, 10, 13, 15, and 19-27. Claim 27 includes the amino acid sequence described in any one of SEQ ID NOs: 19 to 27. The tumor antigen peptide described.

29. The non-protein coding region of the genome is an intron, and the tumor antigen peptide D is one of sequence numbers 16, 17, and 36-64, preferably sequence numbers 36-64. The tumor antigen peptide according to claim 27, comprising any one of the amino acid sequences described herein.

30. The non-protein coding region of the genome is an intergenetic region, and the tumor antigen peptide Claims that the cido comprises the amino acid sequence described in any one of SEQ ID NOs. 65 to 84. The tumor antigen peptide described in 27.

31. The non-protein coding region of the genome is a non-coding RNA transcript (ncRNA) The exon is the tumor antigen peptide, and the tumor antigen peptide is preferably the sequence numbers 4 and 85-92. Claim 27, comprising the amino acid sequence described in any one of the row numbers 85 to 92 Tumor antigen peptide.

32. The non-protein coding region of the genome is the antisense strand of the gene, and the genome The ulcer antigen peptide contains the amino acid sequence described in any one of SEQ ID NOs: 93-99 The tumor antigen peptide according to claim 27.

33. A nucleic acid encoding a tumor antigen peptide according to any one of claims 1 to 32.

34. The nucleic acid according to claim 33, which is mRNA or a viral vector.

35. The tumor antigen peptide according to any one of claims 1 to 32, or claim 33 or A liposome containing the nucleic acid described in 34.

36. The tumor antigen peptide according to any one of claims 1 to 32, or claim 33 or 34 The nucleic acid described, or the liposome described in claim 35, and a pharmaceutically acceptable carrier are included. Hmm, composition.

37. The tumor antigen peptide according to any one of claims 1 to 32, or claim 33 or 34 The nucleic acid described, the liposome described in claim 35, or the composition described in claim 36, and A vaccine containing an adjuvant.

38. The peptide bond groove contains the tumor antigen peptide according to any one of claims 1 to 32. Hmm, an isolated major histocompatibility complex (MHC) class I molecule.

39. An isolated MHC class I molecule according to claim 38, in the form of a polymer.

40. The isolated MHC class I molecule according to claim 39, wherein the polymer is a tetramer.

41. (i) the tumor antigen peptide according to any one of claims 1 to 32, or (ii) claim The nucleotide sequence comprises a nucleotide sequence encoding a tumor antigen peptide as described in any one of items 1 to 32. Isolated cells containing a vector.

42. The tumor antigen peptide according to any one of claims 1 to 32 within those peptide binding grooves A major histocompatibility complex (MHC) class I molecule containing is expressed on its surface, isolated cells that were processed.

43. The cell according to claim 42, which is an antigen-presenting cell (APC).

44. The cell according to claim 43, wherein the APC is a dendritic cell.

45. An isolated MHC class I molecule and / or as described in any one of claims 38 to 40. MHC class I expressed on the surface of the cell according to any one of claims 42 to 44 T cell receptors (TCRs) are molecules that specifically recognize other molecules.

46. An isolated CD8 expresses the TCR described in claim 45 on its cell surface. ＋ T Lymphocytes.

47. At least 0.5% of the CD8 defined in claim 46 ＋ A group of cells, including T lymphocytes.

48. A method for treating ovarian cancer in the subject, wherein an effective amount of (i) Claims 1 to 32 (ii) the tumor antigen peptide according to any one of the claims, (ii) the nucleic acid according to claim 33 or 34 (iii) the liposome according to claim 35, (iv) the composition according to claim 36, (v) (vi) the vaccine according to claim 37, (vi) the cell according to any one of claims 41 to 45 (vii) CD8 according to claim 46 ＋ T lymphocytes, or (viiii) as described in claim 47 A method comprising administering the cell population described above to the subject.

49. The method according to claim 48, wherein the ovarian cancer is serous carcinoma.

50. The method according to claim 49, wherein the serous carcinoma is high-grade serous carcinoma (HGSC). 。

51. Further comprising administering at least one additional antitumor agent or therapy to the subject. or the method according to any one of claims 48 to 50.

52. The aforementioned at least one additional antitumor agent or therapy is a chemotherapy agent, immunotherapy, or immunotherapy agent. The method according to claim 51, wherein the method is a block point inhibitor, radiotherapy, or surgery.

53. (i) the tumor antigen peptide according to any one of claims 1 to 32, (ii) claim 33 or the nucleic acid according to claim 34, (iii) the liposome according to claim 35, (iv) The composition according to claim 36, (v) the vaccine according to claim 37, (vi) the composition according to claims 41 to 45 (vii) the cells described in any one of the claims, (vii) the CD8 described in claim 46 ＋ T lymphocytes, or (viiii) A cell population according to claim 47 for treating ovarian cancer in a subject, use.

54. (i) the tumor antigen peptide according to any one of claims 1 to 32, (ii) claim 33 or the nucleic acid according to claim 34, (iii) the liposome according to claim 35, (iv) The composition according to claim 36, (v) the vaccine according to claim 37, (vi) the composition according to claims 41 to 45 (vii) the cells described in any one of the claims, (vii) the CD8 described in claim 46 ＋ T lymphocytes, or (viiii) A cell population according to claim 47 for treating ovarian cancer in a subject Use for the manufacture of pharmaceuticals.

55. The use according to claim 53 or 54, wherein the ovarian cancer is serous carcinoma.

56. The use according to claim 55, wherein the serous carcinoma is high-grade serous carcinoma (HGSC). 。

57. Claims 53 to 56 further include the use of at least one additional antitumor agent or therapy. Use as described in any one of the items.

58. The aforementioned at least one additional antitumor agent or therapy is a chemotherapy agent, immunotherapy, or immunotherapy agent. The use according to claim 57, which is a block point inhibitor, radiotherapy, or surgery.

59. (i) For use in treating ovarian cancer in the subject, claims 1 to 32 (ii) the tumor antigen peptide according to any one of the claims, (ii) the nucleic acid according to claim 33 or 34 (iii) the liposome according to claim 35, (iv) the composition according to claim 36, (v (vi) the vaccine according to claim 37, (vi) the cell according to any one of claims 41 to 45 (vii) CD8 according to claim 46 ＋ T lymphocytes, or (viiii) in claim 47 The cell population described.

60. The tumor antigen peptide for use according to claim 59, wherein the ovarian cancer is serous carcinoma. Nucleic acid, liposome, composition, vaccine, cell, CD8 ＋ T lymphocytes, or cell populations 。

61. The use according to claim 60, wherein the serous carcinoma is high-grade serous carcinoma (HGSC). Tumor antigen peptides, nucleic acids, liposomes, compositions, vaccines, cells, CD8 ＋ Tri A cell, or a group of cells.

62. The tumor antigen peptide, nucleic acid, liposome, composition, vaccine, cell, CD8 ＋ T lymphocyte Pacocytes, or cell populations, are used in combination with at least one additional antitumor agent or therapy. Tumor antigen peptides, nucleic acids, liposomes for use as described in any one of items 59 to 61. M, composition, vaccine, cells, CD8 ＋ T lymphocytes, or a group of cells.

63. The aforementioned at least one additional antitumor agent or therapy is a chemotherapy agent, immunotherapy, or immunotherapy agent. A tumor for use according to claim 62, which is a tumor-causing inhibitor, radiotherapy, or surgery. Cerebral antigen peptide, nucleic acid, liposome, composition, vaccine, cell, CD8 ＋ T lymphocytes, A group of cells.

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