CN111863130A

CN111863130A - Screening method and application of tumor immunotherapy prognosis marker

Info

Publication number: CN111863130A
Application number: CN202010650063.7A
Authority: CN
Inventors: 赵松辉; 宋越强; 王凯
Original assignee: Origimed Technology Shanghai Co ltd
Current assignee: Origimed Technology Shanghai Co ltd
Priority date: 2020-04-07
Filing date: 2020-07-08
Publication date: 2020-10-30

Abstract

The invention relates to a screening method of tumor immunotherapy prognosis markers, which comprises the following steps: acquiring expression data, mutation data and clinical curative effect data of a candidate gene group aiming at a cancer species to be analyzed; performing statistical independence test on the mutation condition of the candidate gene group and the expression condition of an immune genome, finding out the gene mutation related to the immune genome, and sequencing the genes in the candidate gene group from high to low according to the degree of association; wherein the immune genome comprises CD8A, GZMA, GZMB, IFN γ, EOMES, CXCL9, CXCL10, and TBX 21; dividing the first N genes into a mutation group and a non-mutation group according to whether the first N genes are mutated or not, performing survival analysis, and selecting M genes with the most obvious difference according to the change of log-rank for reservation; the resulting genes were validated in independent datasets to determine their mutation and prognostic relevance.

Description

Screening method and application of tumor immunotherapy prognosis marker

RELATED APPLICATIONS

The priority of the chinese patent application entitled "method for screening prognostic markers for tumor immunotherapy and use", filed on 07/04/2020, having application number 202010265655.7, is hereby incorporated by reference in its entirety.

Technical Field

The invention relates to the field of medical diagnostics, in particular to a screening method and application of a tumor immunotherapy prognosis marker.

Background

In recent years, tumor immunotherapy, especially immune checkpoint inhibitors, bring long-term survival benefit to various tumor patients, currently PD-L1(programmed death-ligand 1) is the most widely used immunotherapy biomaker, and overexpression of tumor cells PD-L1 can inhibit the function of T cells and enable tumor cells to escape immunologically. A plurality of clinical trials and a plurality of analysis indexes (ORR, PFS and OS) also suggest that the expression of PD-L1 can predict the curative effect of I-O (immune-immune) immunotherapy, and the high expression of PD-L1 is positively correlated with clinical benefit. However, other studies have shown that patients who are PD-L1 negative in lung cancer may also benefit from anti-PD-L1 therapy, and similar phenomena occur in other tumor species. Meanwhile, the detection of PD-L1 is influenced by various factors, such as intratumoral heterogeneity and intratumoral heterogeneity, non-uniform detection standards, different tissue sources and the like. Therefore, there is a limitation in predicting the therapeutic effect of immunotherapy based on the expression amount of a single gene as biorarker.

TMB (Tumor mutation Burden) is also a widely used biomaker predictive of immunotherapy, where TMB is the total number of mutations in coding regions of the Tumor genome and Tumor cells with higher levels of TMB are more readily recognized by the immune system to elicit an immune response. A plurality of large-scale clinical tests such as CheckMate-026, CheckMate-227 and the like have proved the clinical application value of TMB, and the higher the TMB is, the more the clinical benefit is. However, TMB has certain limitations as a biomarker for predicting immunotherapy, for example, the types of gene mutations include point mutation, insertion deletion mutation, copy number variation, etc., and the prediction meanings of different mutation types are different, so a professional detection mechanism is required to evaluate the weights of different types of mutations through analysis of a large amount of data. In addition, the TMB is detected by using whole exon sequencing, so that the detection period is long, the economic cost is high, and the like, and besides, the cutoff value of the TMB is the same as the expression of PD-L1, and the unified standard is not provided.

In view of the above, the present invention is particularly proposed.

Disclosure of Invention

In order to solve the above problems, the inventors have established a novel method for detecting a marker that can predict the therapeutic effect of immunotherapy in different cancer species by combining the expression level of a specific immune gene and gene mutation. The inventor selects 8 immunity-related genes (CD8A, GZMA, GZMB, IFN gamma, EOMES, CXCL9, CXCL10 and TBX21), and finds out immunity-related gene mutation according to the expression level of the 8 immunity genes. And predicting the curative effect of the immunotherapy according to whether the gene is mutated or not. In the application process, the immunotherapy curative effect can be predicted only by detecting whether a limited number of specific genes are mutated, so that the problems of low flux, high cost and the like caused by widely used biorarkers such as PD-L1 and TMB can be well solved, and the problems of non-uniform standard, tumor heterogeneity and the like of the existing biorarkers exist.

Specifically, the invention relates to a screening method of a tumor immunotherapy prognosis marker, which comprises the following steps:

a) acquiring expression data, mutation data and clinical curative effect data of a candidate gene group aiming at a cancer species to be analyzed;

b) performing statistical independence test on the mutation condition of the candidate gene group and the expression condition of an immune genome, finding out the gene mutation related to the immune genome, and sequencing the genes in the candidate gene group from high to low according to the degree of association;

wherein the immune genome comprises CD8A, GZMA, GZMB, IFN γ, EOMES, CXCL9, CXCL10, and TBX 21;

c) dividing the first N genes into a mutation group and a non-mutation group according to whether the first N genes are mutated or not, performing survival analysis, and selecting M genes with the most obvious difference according to the change of log-rank for reservation;

d) validating the genes obtained in step c) in an independent dataset to determine their mutation and prognosis relevance.

The present invention also relates to a computer-readable storage medium associated with the above method for storing a computer instruction, a program, a set of codes or a set of instructions which, when run on a computer, causes the computer to perform the method for screening for prognostic markers for tumor immunotherapy as described above.

The invention also relates to an electronic device associated with the above method, comprising:

one or more processors; and

a storage device storing one or more programs,

when executed by the one or more processors, cause the one or more processors to implement a method for screening for a prognostic marker for tumor immunotherapy as described above.

According to still another aspect of the present invention, the invention also relates to the application of the detection agent of the marker obtained by screening the screening method of the tumor immunotherapy prognosis marker as described above in the preparation of a kit for the recurrence risk assessment, prognosis judgment, treatment or adjuvant therapy of tumors.

The invention has the beneficial effects that:

1. based on biological and immunological principles, the immunogenicity of tumor cells is assessed by the expression levels of key immune genes (CD8A, GZMA, GZMB, IFN γ, EOMES, CXCL9, CXCL10, TBX21) associated with the activation of effector T cells in the immune response;

2. based on the expression level of 8 key immune genes, relevant gene mutations are found to evaluate the gene mutations affecting the immunogenicity of tumor cells;

3. based on gene mutation, the tumor is predicted to activate autoimmunity after an immune checkpoint inhibitor is used, and a good treatment effect is achieved.

In summary, the method can simplify the detection content, reduce the detection cost, accelerate the detection time, and eliminate the interference caused by different platforms, different standards, different thresholds, and the like.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.

FIG. 1 is a schematic flow chart of a method for screening prognostic markers for tumor immunotherapy, which is used in one embodiment of the present invention;

FIG. 2 is a graph showing the results of the validation of markers screened for colon cancer in one embodiment of the present invention.

Detailed Description

Reference will now be made in detail to embodiments of the invention, one or more examples of which are described below. Each example is provided by way of explanation, not limitation, of the invention. In fact, it will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the scope or spirit of the invention. For instance, features illustrated or described as part of one embodiment, can be used on another embodiment to yield a still further embodiment.

It is therefore intended that the present invention cover the modifications and variations of this invention provided they come within the scope of the appended claims and their equivalents. Other objects, features and aspects of the present invention are disclosed in or are apparent from the following detailed description. It is to be understood by one of ordinary skill in the art that the present discussion is a description of exemplary embodiments only, and is not intended as limiting the broader aspects of the present invention.

The invention relates to a screening method of tumor immunotherapy prognosis markers, which comprises the following steps:

Mutations in important regulatory genes (tumor suppressor genes tumor supressors and proto-oncogenes) alter the behavior pattern of cells and may lead to the development of malignant tumors. All tumors must be controlled by a series of signal pathways, and the related genes may be different from each other and the functions of the genes are different. Even if two different patients suffer from the same cancer species, the involved tumor suppressor genes and oncogenes may differ greatly. Whereas proteins are the basis for the biological function of tumors and are synthesized based on the transcription of mRNA, mutations in the DNA of the coding region result in alterations in the mRNA. The mRNA changes may in turn lead to the formation of dysfunctional proteins.

Mutations include a variety of types, each of which affects mRNA expression. Even a single nucleotide (nucleotide) change in the DNA strand may result in the failure of mRNA transcription and the complete loss of protein function. If the oncogene (oncogene) is located in the amplification region, overexpression of the gene (overexpression) can lead to uncontrolled cell growth. For example, amplification of myc oncogenes in many tumors, and amplification of ErbB-2 or HER-2/neu oncogenes in breast and ovarian cancers. Currently, clinical treatment of the HER-2/neu oncogene is directed primarily to those cells that overexpress the protein product.

Under normal conditions, a large number of mutations are accumulated in the development process of tumors, the mutations encode specific tumor-associated antigens and are released by tumor cells, Antigen Presenting Cells (APCs) recognize, take up and process tumor antigens, are combined with MHC (major histocompatibility complex) molecules (including CD8A molecules) and present on the cell surface, ACPs enter lymphatic tissues, and Src kinase LCK is recruited to the vicinity of TCR-CD3 complex. LCK initiates different intracellular signaling pathways by phosphorylating various substrates, ultimately leading to the production of lymphokines, activation of Cytotoxic T Lymphocytes (CTL), secretion of GZMA, GZMB, GZMC, PRF1 (granzymes a, B, C and perforin), killing of tumor cells, production of IFN γ, stimulation of chemokines that attract T cells by IFN γ (CXCL9, CLCL10 and CXCL11), limiting the anti-regulatory activity of the immune response, whereas TBX21 protein is a Th1(T helper) cell-specific transcription factor, regulating expression of the Th1 cytokine interferon- γ (IFNG), EOMES is an important paralogue of TBX 21. These 8 immune genes (CD8A, GZMA, GZMB, IFN γ, EOMES, CXCL9, CXCL10, TBX21) are highly co-expressed and highly correlated with T cell activation and interferon γ (the specific functions of the 8 genes are shown in table 1), co-participating in immune surveillance, the process of killing tumor cells.

Although tumor cells express certain specific antigens and are recognized by the immune system, these tumor cells can function to suppress the immune system through the expression of other molecules (e.g., immune checkpoint, PD-1/PD-L1). In this case, although the immune system can recognize tumor cells, it cannot be activated effectively and cannot exert the effect of killing tumor cells. In response to the loss of immunogenicity of tumor cells, Immune Checkpoint Inhibitors (ICIs) immunotherapy, typified by PD-1/PD-L1 monoclonal antibody, are now used in a large number of patients. The ICIs can inhibit the binding of PD-1 on the surface of the T cell and PD-L1 ligand on the surface of the tumor cell, reactivate the T cell and play a role in killing the tumor cell. Therefore, according to 8 genes closely related to T cell activation, the mutation closely related to T cell activation is analyzed in multiple cancer types, and a patient with the mutation can be activated again in ICIs to inhibit and kill tumor cells and prolong the survival time.

The information of 8 genes in the immune genome selected by the invention is shown in table 1:

TABLE 1

In the present invention, a "tumor" may be from a different cancer species, and may be benign or malignant, including, for example: bone, bone junction, muscle, lung, trachea, heart, spleen, artery, vein, blood, capillary vessel, lymph node, lymphatic vessel, lymph fluid, oral cavity, pharynx, esophagus, stomach, duodenum, small intestine, colon, rectum, anus, appendix, liver, gallbladder, pancreas, parotid gland, sublingual gland, urinary kidney, ureter, bladder, urethra, ovary, fallopian tube, uterus, vagina, vulva, scrotum, testis, vas deferens, penis, eye, ear, nose, tongue, skin, brain, brainstem, medulla oblongata, spinal cord, cerebrospinal fluid, nerve, thyroid, parathyroid, adrenal gland, pituitary, pineal gland, pancreatic islet, thymus, gonad gland, sublingual gland, and parotid gland.

In the present invention, the genetic variation may include point mutation (point mutation) and fragment mutation (fragmentmutation); the point mutation may be a Single Nucleotide Polymorphism (SNP), a base substitution, a single base insertion or base deletion, or a silent mutation (e.g., a synonymous mutation); the fragment mutation may be an insertion mutation, a truncation mutation or a gene rearrangement mutation.

In some embodiments, step c) further comprises:

and performing cluster analysis on the M genes, removing redundancy according to variation similarity, and reserving representative genes in each class.

In some embodiments, the method of statistical independence test in step b) is a Fisher test.

In some embodiments, the indicator according to which the degree of association is ranked from high to low in step b) is an OR value.

In some embodiments, the expression level of the immune genome in step b) is the median of the expression levels of the individual genes in the immune genome.

In some embodiments, the immunotherapy is an immune checkpoint inhibitor therapy.

As used herein, the term "immune checkpoint" refers to some inhibitory signaling pathway present in the immune system. Under normal conditions, the immune checkpoint can maintain immune tolerance by adjusting the strength of autoimmune reaction, however, when the organism is invaded by tumor, the activation of the immune checkpoint can inhibit autoimmunity, which is beneficial to the growth and escape of tumor cells. By using the immune checkpoint inhibitor, the normal anti-tumor immune response of the body can be restored, so that the tumor can be controlled and eliminated.

Immune checkpoints according to the invention include, but are not limited to, programmed death receptor 1(PD1), PD-L1, cytotoxic T lymphocyte-associated antigen 4 (CTLA-4); also included are some newly discovered immune checkpoints such as lymphocyte activation gene 3(LAG3), CD160, T cell immunoglobulin and mucin-3 (TIM-3), T cell activated V domain immunoglobulin inhibitor (VISTA), adenosine A2a receptor (A2aR), and the like.

Preferred immune checkpoint inhibitors are PD1 inhibitors and/or PD-L1 inhibitors.

The PD1 inhibitor may further be selected from one or more of Nivolumab (OPDIVO; BMS-936558), Pembrolizumab (MK-3475), Jembrolizumab, lambrolizumab, Pidilizumab (CT-011) Terepril mab (JS001), and Iplilimumab.

The PD-L1 inhibitor may further be selected from one or more of Atezolizumab (MPDL3280A), JS003, Durvalumab, Avelumab, BMS-936559, MEDI4736 and MSB001071 0010718C.

The present invention also relates to a computer-readable storage medium for storing a computer instruction, a program, a set of codes or a set of instructions which, when run on a computer, causes the computer to perform the method for screening for a prognostic marker for tumor immunotherapy as described above.

Any combination of one or more computer-readable media may be employed. The computer readable medium may be a computer readable signal medium or a computer readable storage medium. A computer readable storage medium may be, for example, but not limited to, an electronic, magnetic, optical, electromagnetic, infrared, or semiconductor system, apparatus, or device, or any combination of the foregoing. More specific examples (a non-exhaustive list) of the computer readable storage medium would include the following: an electrical connection having one or more wires, a portable computer diskette, a hard disk, a Random Access Memory (RAM), a read-only memory (ROM), an erasable programmable read-only memory (EPROM or flash memory), an optical fiber, a portable compact disc read-only memory, an optical storage device, a magnetic storage device, or any suitable combination of the foregoing. In the context of this document, a computer readable storage medium may be any tangible medium that can contain, or store a program for use by or in connection with an instruction execution system, apparatus, or device.

A computer readable signal medium may include a propagated data signal with computer readable program code embodied therein, for example, in baseband or as part of a carrier wave. Such a propagated data signal may take any of a variety of forms, including, but not limited to, electro-magnetic, optical, or any suitable combination thereof. A computer readable signal medium may also be any computer readable medium that is not a computer readable storage medium and that can communicate, propagate, or transport a program for use by or in connection with an instruction execution system, apparatus, or device.

Program code embodied on a computer readable medium may be transmitted using any appropriate medium, including but not limited to wireless, wireline, optical fiber cable, RF, etc., or any suitable combination of the foregoing.

Computer program code for carrying out operations for aspects of the present disclosure may be written in any combination of one or more programming languages, including an object oriented programming language such as Java, Smalltalk, C + +, and conventional procedural programming languages, such as the "C" programming language or similar programming languages. The program code may execute entirely on the user's computer, partly on the user's computer, as a stand-alone software package, partly on the user's computer and partly on a remote computer or entirely on the remote computer or server. In the case of a remote computer, the remote computer may be connected to the user's computer through any type of network, including a Local Area Network (LAN) or a Wide Area Network (WAN), or the connection may be made to an external computer (for example, through the Internet using an Internet service provider).

According to yet another aspect of the invention, it also relates to an electronic device comprising:

One or more processors; and

a storage device storing one or more programs,

Optionally, the electronic device may further comprise a transceiver. The processor is coupled to the transceiver, such as via a bus. It should be noted that the transceiver in practical application is not limited to one, and the structure of the electronic device does not constitute a limitation to the embodiments of the present application.

The processor may be a CPU, general purpose processor, DSP, ASIC, FPGA or other programmable logic device, transistor logic device, hardware component, or any combination thereof. Which may implement or perform the various illustrative logical blocks, modules, and circuits described in connection with the disclosure. A processor may also be a combination of computing functions, e.g., comprising one or more microprocessors, a DSP and a microprocessor, or the like.

A bus may include a path that transfers information between the above components. The bus may be a PCI bus or an EISA bus, etc. The bus may be divided into an address bus, a data bus, a control bus, etc. The memory 802 may be, but is not limited to, a ROM or other type of static storage device that can store static information and instructions, a RAM or other type of dynamic storage device that can store information and instructions, an EEPROM, a CD-ROM or other optical disk storage, optical disk storage (including compact disk, laser disk, optical disk, digital versatile disk, blu-ray disk, etc.), a magnetic disk storage medium or other magnetic storage device, or any other medium that can be used to carry or store desired program code in the form of instructions or data structures and that can be accessed by a computer.

In other embodiments, the invention also relates to the application of the detection agent of the marker obtained by screening the screening method of the tumor immunotherapy prognosis marker in preparing a kit for the recurrence risk assessment, prognosis judgment, treatment or adjuvant treatment of tumors.

In some embodiments, the marker comprises one or more of FLT1, STK11, EPHA5, PRKD1, HGF, FAT1, ARID1B, inp 4B, EPHB1, and MTOR.

In some embodiments, the markers include one or more of FLT1, STK11, EPHA5, PRKD1, HGF, FAT1, ARID1B, inp 4B, EPHB1, and MTOR for risk of recurrence assessment, prognostic judgment, treatment, or adjuvant treatment of colon cancer.

The above-mentioned detection agent is used for judging the mutation of the marker, and thus the detection can be carried out at the DNA level, the RNA level (if the marker is transcribed) or the protein level (if the marker is translated).

As the detection agent for a nucleic acid level (DNA or RNA level), a known agent known to those skilled in the art can be used, for example, a nucleic acid (usually a probe or primer) which can hybridize to the DNA or RNA and is labeled with a fluorescent label, and the like. And one skilled in the art would also readily envision reverse transcribing mRNA into cDNA and detecting the cDNA, and routine replacement of such techniques would not be outside the scope of the present invention.

In some embodiments, the detection agent is used to perform any one of the following methods:

polymerase chain reaction, denaturing gradient gel electrophoresis, nucleic acid sequencing, nucleic acid typing chip detection, denaturing high performance liquid chromatography, in situ hybridization, biological mass spectrometry and HRM method.

In some embodiments, the polymerase chain reaction is selected from the group consisting of restriction fragment length polymorphism, single strand conformation polymorphism, Taqman probe, competitive allele-specific PCR, and allele-specific PCR.

In some embodiments, the biomass spectrometry is selected from flight mass spectrometer detection.

In some embodiments, the nucleic acid sequencing method is selected from the Snapshot method.

In some embodiments of the invention, the nucleic acid sequencing method may be transcriptome sequencing or genome sequencing. In some further embodiments of the invention, the nucleic acid sequencing method is high throughput sequencing, also known as next generation sequencing ("NGS"). Second generation sequencing produces thousands to millions of sequences simultaneously in a parallel sequencing process. NGS is distinguished from "Sanger sequencing" (one generation sequencing), which is based on electrophoretic separation of chain termination products in a single sequencing reaction. Sequencing platforms that can be used with the NGS of the present invention are commercially available and include, but are not limited to, Roche/454 FLX, Illumina/Solexa genome Analyzer, and Applied Biosystems SOLID system, among others. Transcriptome sequencing can also rapidly and comprehensively obtain almost all transcripts and gene sequences of a specific cell or tissue of a certain species in a certain state through a second-generation sequencing platform, and can be used for researching gene expression quantity, gene function, structure, alternative splicing, prediction of new transcripts and the like.

In some embodiments, the detection agent is detected at the protein level.

biological mass spectrometry, amino acid sequencing, electrophoresis, and detection using antibodies specifically designed for the mutation site.

The detection method using an antibody specifically designed for the mutation site may further be immunoprecipitation, co-immunoprecipitation, immunohistochemistry, ELISA, Western Blot, or the like.

The invention also relates to a method for assessing the risk of recurrence, prognostically judging, treating or adjunctively treating a tumor, which comprises:

detecting a mutation in a marker from a biological sample obtained from the subject and evaluating the patient;

the marker is obtained by screening the tumor immunotherapy prognosis marker by the screening method.

The biological sample can be any sample obtained from a subject, e.g., tissue, cell, body fluid, among others. Preferably, the biological sample is tissue, blood, plasma, serum, whole blood, urine, saliva, genital secretions, cerebrospinal fluid, sweat, faeces or bronchoalveolar lavage fluid. More preferably, the tissue is lung tissue, further may be lung cancer tissue.

Wherein the step of evaluating may be performed on a computer system.

Embodiments of the present invention will be described in detail with reference to examples.

Examples

In a representative embodiment, the method provided by the present invention involves 6 steps,

1. through Fisher's test, gene mutations associated with expression of 8 immune genes were found.

2. Ranking according to the degree of association between the mutant genes and the expression of 8 immune genes (i.e., ranking according to the OR value of the mutant genes, the higher the OR, the greater the probability of the gene mutation occurring when the gene is expressed at a high level)

3. Dividing the first N genes into a mutation group and a non-mutation group according to whether the first N genes are mutated or not, performing log-rank test, and analyzing the life cycle difference.

4. Of the first M genes, the differences in survival were most significant (i.e., log-rank test, p-value was minimal). To further reduce dimension, the M genes can be clustered into n classes.

5. Correlation analysis is performed on each class (cluster) in the n classes. Selecting representative gene in each class (i.e., the gene has the largest correlation coefficient with other genes in cluster)

6. Finally, n genes are reserved, and whether the obtained genes can be used for characterizing the life cycle difference or not is verified in an independent data set.

The method is suitable for finding markers for predicting the efficacy of immunotherapy in a plurality of cancer species, and when the method is used to study a specific cancer species, the cancer species data is required to include expression data (including 8 immune genes), mutation data, and clinical efficacy data (such as OS, PFS). In this example, the data for the candidate genes are from the TCGA dataset and the independent dataset used in the validation at step 6 is the MSK dataset.

By analyzing the data by the method, N genes can be found for predicting the curative effect of immunotherapy. And finally, the kit can be clinically applied, and the curative effect of immunotherapy is judged by only detecting whether the N genes are mutated, so that clinical guidance is provided.

The flow diagram of the above method is shown in fig. 1.

In this example, analysis was performed for Colon Cancer (COAD), and finally 10 genes (FLT1, STK11, EPHA5, PRKD1, HGF, FAT1, ARID1B, inp 4B, EPHB1, MTOR) were found as biomarkers for predicting immunotherapy. In all data validation using the external data set MSK, the mutation group (MT) was more sensitive to immunotherapy and survived significantly more than the wild group (wild type, WT) (see fig. 2). Among them, 10 genes were used as a mutation group as long as at least one of the genes was mutated.

The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.

The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.

Claims

1. A method for screening prognostic markers for tumor immunotherapy, comprising:

2. The method for screening prognostic markers for tumor immunotherapy according to claim 1, wherein the step c) further comprises:

3. The method for screening prognostic markers for tumor immunotherapy according to claim 1, wherein the statistical independence test in step b) is Fisher test.

4. The method for screening prognostic markers for tumor immunotherapy according to claim 3, wherein the index according to which the degree of association is ranked from high to low in step b) is an OR value.

5. The method for screening prognostic markers for tumor immunotherapy according to claim 1, wherein the expression level of the immune genome in step b) is the median of the expression levels of the individual genes in the immune genome.

6. The method for screening prognostic markers for immunological treatment of tumors according to any one of claims 1 to 5, wherein the immunological treatment is an immune checkpoint inhibitor therapy.

7. The screening method for prognostic markers for tumor immunotherapy according to claim 6, wherein the immune checkpoint inhibitor is a PD1 inhibitor and/or a PD-L1 inhibitor.

8. A computer readable storage medium for storing a computer instruction, program, code set or instruction set which, when run on a computer, causes the computer to perform a method of screening for a prognostic marker for tumour immunotherapy as defined in any one of claims 1 to 7.

9. An electronic device, comprising:

one or more processors; and

a storage device storing one or more programs,

when executed by the one or more processors, cause the one or more processors to implement the method for screening for a prognostic marker for tumor immunotherapy as defined in any one of claims 1 to 7.

10. Use of the marker detection agent obtained by screening the tumor immunotherapy prognosis marker according to any one of claims 1 to 7 in the preparation of a kit for tumor recurrence risk assessment, prognosis, therapy or adjuvant therapy.

11. The use according to claim 10, comprising one or more of FLT1, STK11, EPHA5, PRKD1, HGF, FAT1, ARID1B, inp 4B, EPHB1 and MTOR.

12. The use of claim 11, wherein the tumor is colon cancer.