Use of IFP35 protein family in evaluation of efficacy of tumor immunotherapy

Abstract

The present invention relates to the use of the IFP35 protein family in the evaluation of the efficacy of tumor immunotherapy. The present invention discloses, for the first time, that the IFP35 protein family (interferon-induced 35 kDa protein (IFP35) and / or N-Myc interacting protein (NMI)) can be used as a marker for evaluating the efficacy of tumor immunotherapy, evaluating the prognosis of tumor immunotherapy and / or selecting a treatment scheme, and by means of the IFP35 protein family (interferon-induced 35 kDa protein (IFP35) and / or N-Myc interacting protein (NMI)), and / or an IFP35 protein family (interferon-induced 35 kDa protein (IFP35) and / or N-Myc interacting protein (NMI)) promoter, the efficacy of tumor immunotherapy can be improved.

Description

Application of IFP35 protein family in evaluating the efficacy of tumor immunotherapy Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to the application of the IFP35 protein family in evaluating the efficacy of tumor immunotherapy. Background Art

[0002] Cancer is one of the most serious diseases that threaten human life and health. Currently, the commonly used cancer treatment methods mainly include surgery, chemotherapy, radiotherapy and targeted therapy, which have made great progress and greatly improved the survival of cancer patients. The initial clinical treatment effect of many tumors is good, but as the treatment time prolongs, the tumor often develops drug resistance and is prone to recurrence, which brings severe challenges to clinical treatment. Tumor immunotherapy is a new type of therapy that has developed rapidly in the past decade. Its main target is the body's immune system rather than tumor cells. It eliminates tumor cells by enhancing the body's natural immune defense against tumors and reshaping the immune microenvironment. On the one hand, these therapies train immune cells to recognize and eliminate target cells carrying tumor antigens, enhance immune-mediated tumor cell lysis, and on the other hand, eliminate or reduce the immunosuppressive signals induced by tumor cells, thereby achieving the effect of tumor treatment [1]. In December 2013, the American magazine Science ranked tumor immunotherapy as the top ten scientific breakthroughs of the year. The 2018 Nobel Prize in Physiology or Medicine was awarded to tumor immunotherapy. The main immunotherapies currently under clinical research and application include monoclonal antibody therapy, immune checkpoint inhibitors (ICIs) therapy, adoptive cell therapy (ACT), oncolytic virus therapy (OVT) and tumor vaccine therapy.

[0003] Monoclonal antibody therapy: Monoclonal antibodies consist of a Fab end that binds to tumor antigens and an Fc end that binds to immune cell surface receptors. Under the action of the monoclonal antibody, the two cells bind and kill the tumor cells through complement-mediated cytotoxicity and antibody-dependent cytotoxicity. Another monoclonal antibody-based therapy, antibody-drug conjugates (ADCs), are targeted biologics that are coupled to highly cytotoxic drugs through specific linkers. They can specifically bind to tumor surface antigens and release drugs through a "self-immolation" mechanism to kill tumor cells and activate the immune system [2].

[0004] Immune checkpoint inhibitor therapy: Immune checkpoints are protective factors in the body's immune system that can prevent T cells from being overactivated and causing autoimmune damage. However, tumor cells may exploit these checkpoints to escape the body's immune surveillance and killing, resulting in immune escape. Immune checkpoint inhibitors can effectively restore the function of T cells by blocking immune checkpoints. The 2018 Nobel Prize in Physiology or Medicine was awarded to scientists who made great contributions to the research of two classic checkpoints, CTLA-4 and PD-1. As research deepens, more and more immune checkpoints have been discovered, such as LAG3, TIGIT, TIM3, adenosine A2A and CD47, and drug development targeting these newly discovered immune checkpoints is also ongoing [3].

[0005] Adoptive cell therapy: Adoptive cell therapy is a passive immunotherapy method. First, immune cells are isolated from the tumor patient, genetically engineered or screened and activated in vitro, and then amplified in large quantities and then reinfused into the patient to achieve the purpose of clearing the tumor. Depending on whether the effector cells express exogenous genes, adoptive cell therapy can be divided into two categories: one requires genetic engineering, such as chimeric antigen receptor T cells (CAR-T) and T cell receptor chimeric T cells (TCR-T) therapy that genetically modify T cells to express chimeric antigen receptors (CAR) or T cell receptors (TCR), and CAR-NK therapy that expresses CAR on the surface of NK cell membranes; the other is to isolate and screen immune cells from the patient's peripheral blood or tumor in situ, amplify and activate in vitro, and then reinfuse them into the patient's body for anti-tumor treatment, including tumor infiltrating immune cells (TILs) therapy, cytokine-induced killer (CIK) cell therapy, lymphokine-activated killer (LAK) cell therapy, natural killer (NK) cell therapy, etc. [4].

[0006] Oncolytic virus therapy: Oncolytic viruses (OVs) are naturally occurring or genetically engineered viruses that can selectively replicate in tumor cells, causing tumor cell lysis and exposing antigens, thereby activating the immune system[5].

[0007] Therapeutic tumor vaccine therapy: Therapeutic tumor vaccine therapy is an active immunotherapy. Unlike conventional preventive vaccines, the injection of therapeutic tumor vaccines into patients with malignant tumors will induce or enhance the body's pre-existing humoral and cellular immunity targeting tumor antigens, killing tumor cells while forming long-term immune memory, which can prevent tumor recurrence to a certain extent[6].

[0008] Tumor immunotherapy has made some progress in the treatment of tumors, but there are still problems such as low efficacy and primary drug resistance in some patients. Therefore, it is necessary to study markers for evaluating the efficacy of tumor immunotherapy and methods to improve the efficacy of tumor immunotherapy in clinical practice. 1. Yang, Y., Cancer immunotherapy: harnessing the immune system to battle cancer. J Clin Invest, 2015. 125(9): p. 3335-7.

[0009] 2. Weiner, GJ, Building better monoclonal antibody-based therapeutics. Nat Rev Cancer, 2015.15(6):p.361-70.

[0010] 3. Wei, G., et al., Emerging immune checkpoints in the tumor microenvironment: Implications for cancer immunotherapy. Cancer Lett, 2021.511: p.68-76.

[0011] 4. Wang, Z. and YJCao, Adoptive Cell Therapy Targeting Neoantigens: A Frontier for Cancer Research. Front Immunol, 2020.11: p.176.

[0012] 5. Fukuhara, H., Y. Ino, and T. Todo, Oncolytic virus therapy: A new era of cancer treatment at dawn. Cancer Sci, 2016.107(10):p.1373-1379.

[0013] 6. Song, Q., CDZhang, and XHWu, Therapeutic cancer vaccines: From initial findings to prospects. Immunol Lett, 2018.196: p.11-21.

[0014] Summary of the Invention

[0015] The IFP35 protein family (interferon-induced protein 35kD (IFP35) and N-Myc interacting protein (NMI)) is an inflammatory factor first discovered and identified by the inventors of this application. The inventors of this application have filed a series of patent applications for this purpose: PCT International Application PCT / CN2015 / 000602, Application No. CN202010122554.4, and PCT International Application PCT / CN2020 / 082296; the entire contents of these applications are hereby incorporated herein for all purposes.

[0016] The first aspect of the present invention aims to provide a use of a substance for detecting a marker in a product.

[0017] The second aspect of the present invention aims to provide the use of the IFP35 protein family in the preparation of a tumor immunotherapy enhancer.

[0018] The third aspect of the present invention aims to provide a tumor immunotherapy enhancer.

[0019] The fourth aspect of the present invention aims to provide an anti-tumor pharmaceutical composition.

[0020] The fifth aspect of the present invention aims to provide an anti-tumor drug kit.

[0021] The sixth aspect of the present invention aims to provide a method.

[0022] The seventh aspect of the present invention aims to provide a system.

[0023] The eighth aspect of the present invention aims to provide a marker combination.

[0024] The ninth aspect of the present invention aims to provide a product.

[0025] The purpose of the tenth aspect of the present invention is to provide the use of markers as targets in the development of products related to tumor immunotherapy.

[0026] The eleventh aspect of the present invention aims to provide a tumor immunotherapy enhancer and a tumor immunotherapy agent for use in the preparation of drugs for treating tumors.

[0027] The twelfth aspect of the present invention is to provide a method for improving the efficacy of tumor immunotherapy.

[0028] The thirteenth aspect of the present invention aims to provide a method for treating tumors.

[0029] The purpose of the fourteenth aspect of the present invention is to provide a method for constructing a model with poor tumor immunotherapy efficacy.

[0030] The purpose of the fifteenth aspect of the present invention is to provide a method for constructing a model with good tumor immunotherapy efficacy.

[0031] In order to achieve the above object, the technical solution adopted by the present invention is:

[0032] In a first aspect of the present invention, there is provided use of a substance for detecting a marker in preparing a product, wherein the marker comprises the IFP35 protein family, wherein the IFP35 protein family comprises interferon-induced protein 35kD (IFP35) and / or N-Myc interacting protein (NMI);

[0033] The product is used for at least one of (1)-(3): (1) evaluation of the efficacy of tumor immunotherapy; (2) prognosis evaluation of tumor immunotherapy; (3) selection of treatment regimen.

[0034] Preferably, tumor immunotherapy efficacy evaluation refers to judging the effect of tumor immunotherapy (whether there is a response to tumor immunotherapy) based on the level of markers.

[0035] Preferably, the prognosis assessment of tumor immunotherapy refers to judging the prognosis (good prognosis or poor prognosis) based on the level of the marker.

[0036] Preferably, the treatment regimen selection refers to selecting whether to use a tumor immunotherapy enhancer (tumor immunotherapy, or tumor immunotherapy combined with a tumor immunotherapy enhancer) based on the level of the marker.

[0037] Preferably, the substance for detecting a marker comprises a substance for quantitatively detecting a marker.

[0038] Preferably, the substance for detecting a marker includes a substance for detecting a marker at a gene level and / or a protein level.

[0039] Preferably, the substance comprises a substance for one or more detection techniques or methods selected from the group consisting of immunohistochemistry, Western blotting, Northern blotting, PCR, biochip, nucleic acid sequencing, and amino acid sequencing.

[0040] Preferably, the immunohistochemistry method is selected from at least one of immunofluorescence assay, enzyme-linked immunosorbent assay (ELISA) and immunocolloidal gold assay.

[0041] Preferably, the substance for detecting the marker is selected from one or more of: a substance specific to the marker, a marker-specific probe, a gene chip, and a PCR primer.

[0042] Preferably, the substance specific for the marker comprises any one of (a1) to (a3): (a1) an antibody that specifically binds to the marker; (a2) a ligand protein or polypeptide that specifically binds to the marker; (a3) ​​a non-protein compound that specifically recognizes the marker.

[0043] Preferably, the antibody comprises at least one of a polyclonal antibody, a monoclonal antibody, a single-chain antibody, a functional antibody fragment, an antibody Fab region, a nanobody, a chimeric antibody, and a multispecific antibody.

[0044] Preferably, the product comprises at least one of a reagent, a kit, a test paper, a chip, and a system.

[0045] Preferably, the test sample of the product is selected from at least one of body fluids, tissues, cells, and excretions of the subject to be tested.

[0046] Preferably, the body fluid comprises at least one of blood and lymph.

[0047] Preferably, the blood comprises at least one of serum, plasma, dried blood spots, and whole blood.

[0048] Preferably, the tissue comprises tumor tissue.

[0049] Preferably, the excrement comprises at least one of urine, feces, and tears.

[0050] Preferably, the subject to be tested includes mammals, such as humans, non-human primates (such as gorillas and apes), rodents (such as rats, mice, and guinea pigs), pets (such as cats and dogs), and livestock (such as horses, cows, sheep, pigs, and rabbits).

[0051] Preferably, the subject to be tested includes humans.

[0052] Preferably, the detected level of the IFP35 protein family is positively correlated with the evaluation of tumor immunotherapy efficacy and tumor immunotherapy prognosis, that is, the higher the level of the detected IFP35 protein family, the better the tumor immunotherapy efficacy and tumor immunotherapy prognosis.

[0053] Preferably, the higher the level of the detected IFP35 protein family, the less the need for combined use of a tumor immunotherapy enhancer.

[0054] Preferably, the markers further include other markers for evaluating the efficacy of tumor immunotherapy, evaluating the prognosis of tumor immunotherapy, and / or selecting treatment regimens.

[0055] Preferably, the other markers for evaluating the efficacy of tumor immunotherapy, evaluating the prognosis of tumor immunotherapy, and / or selecting a treatment regimen can be any marker disclosed in the prior art for evaluating the efficacy of tumor immunotherapy, evaluating the prognosis of tumor immunotherapy, and / or selecting a treatment regimen, such as p62 / SQSTM1 disclosed in patent document CN112014564A, the combination (H3Cit, IL-8 and CRP) disclosed in patent document CN113358872A, the combination (VPS29, SNRPD3, PSMB2, VCP, RAB7A, PSMB4, C1or43, GAPDH, ACTB) disclosed in patent document CN110223729A, and the combination (at least 3 of VPS29, SNRPD3, PSMB2, VCP, RAB7A, PSMB4, C1or43, GAPDH, ACTB) disclosed in patent document CN112014564A. The combination disclosed in CN109576365A (SCML4, TTC16, KIR2DL3, ART4, LINC00426, C15orf53, HEATR9, KIR2DL1, TRAT1, IL27, SLFN14, KEL, CXCR2P1, KIR3DL2, GBP7, BTN1A1, IFNG, CLEC6A, LINC00494, ITIH1, CLECL1, GHRL, C4orf50, WDR64, PLA2G2D, CR1L, OR52N4, TTLL6, FCRL6, PIK3CD-AS1, UPK3A, KIF25, LILRA4, RTP5, CLEC4C, KIR2DS4, C9orf139, DBH, FCER2, CCR9, CD40LG, NCR1, CCL25, RHOXF1, GPR174, TIMD4, IL26, AICDA, KCNJ10, FAM129C, WNT1, FAM180B, KLRC3, CN R2, LINC00158, EC9A, NME8, BCORP1, NCR2, IL12B, LILRP2, KCNA2, CCKAR, CLNK, FCRL4, SLC18A1, CRYBA4, TNFRSF13B, SSTR3, CR2, LCNL1, TTC24, ITGAD, IL17REL, DAZL, ZNF80, CXorf65, BEND4, FCAMR, CD19, HSF5, FNDC9, TUSC5, ARRDC5, GTSF1L, ATP1A4, CCL1, BFSP2, FCRL1, IL5RA, AIRE, MS4A1, UNQ6494, FAM177B, NPHS1,ASCL4, FAM216B, FRMD1); the combination disclosed in patent document CN110456042A (MIP-1beta, SDF-1alpha, PD-L1, CD28, TIM-3, RANTES and any multiple of IP-10), the levels of other markers for tumor immunotherapy efficacy evaluation, tumor immunotherapy prognosis evaluation, and / or treatment regimen selection and tumor immunotherapy efficacy evaluation, tumor immunotherapy prognosis evaluation, and / or treatment regimen selection are referred to the above patent documents.

[0056] The second aspect of the present invention provides the use of (c1) and / or (c2) in the preparation of a tumor immunotherapy enhancer;

[0057] (c1) interferon-induced protein 35kD (IFP35), and / or interferon-induced protein 35kD (IFP35) promoter;

[0058] (c2) N-Myc interacting protein (NMI), and / or N-Myc interacting protein (NMI) enhancer.

[0059] Preferably, the interferon-induced protein 35kD (IFP35) promoter comprises at least one of a substance that increases the level of IFP35, a substance that enhances the activity of IFP35, and a substance that delays the metabolism of IFP35.

[0060] Preferably, the IFP35 promoter comprises at least one of: a naturally purified substance, a modified naturally purified substance, a semi-synthetic substance, and a chemically synthesized substance.

[0061] Preferably, the IFP35 promoter comprises at least one of chemical drugs, polypeptide drugs, protein drugs and gene drugs.

[0062] Preferably, the IFP35 promoter comprises: at least one of: biomaterials related to IFP35, exogenous IFP35, nanoparticles carrying genes (IFP35 genes), PEG-modified proteins (IFP35 proteins), protein microspheres (IFP35 proteins), liposome-encapsulated target genes (IFP35 genes) or proteins (IFP35 proteins), extracellular vesicles carrying target genes (IFP35 genes) or proteins (IFP35 proteins), and IFP35 precursor proteins, conjugates, or complexes that can be converted into IFP35 in vivo.

[0063] Preferably, the biological material related to IFP35 comprises at least one of d1) to d16): d1) a nucleic acid molecule encoding IFP35; d2) an expression cassette comprising the nucleic acid molecule of d1); d3) a vector comprising the nucleic acid molecule of d1); d4) a vector comprising the expression cassette of d2); d5) a transgenic cell line comprising the nucleic acid molecule of d1); d6) a transgenic cell line comprising the expression cassette of d2); d7) a transgenic cell line comprising the vector of d3); d8) a transgenic cell line comprising the vector of d4); d9) a microorganism comprising the nucleic acid molecule of d1); d10) a microorganism comprising the expression cassette of d2); d11) a microorganism comprising the vector of d3); d12) a microorganism comprising the vector of d4); d13) a virus comprising the nucleic acid molecule of d1); d14) a virus comprising the expression cassette of d2); d15) a virus comprising the vector of d3); d16) a virus comprising the vector of d4).

[0064] Preferably, the IFP35 is human IFP35.

[0065] Preferably, the N-Myc interacting protein (NMI) promoter comprises at least one of a substance that increases the level of NMI, a substance that enhances the activity of NMI, and a substance that delays the metabolism of NMI.

[0066] Preferably, the NMI promoter comprises at least one of: a naturally purified substance, a modified naturally purified substance, a semi-synthetic substance, and a chemically synthesized substance.

[0067] Preferably, the NMI promoter comprises at least one of chemical drugs, polypeptide drugs, protein drugs and gene drugs.

[0068] Preferably, the NMI promoter comprises: at least one of: NMI-related biomaterials, exogenous NMI, nanoparticles carrying genes (NMI genes), PEG-modified proteins (NMI proteins), protein microspheres (NMI proteins), liposome-encapsulated target genes (NMI genes) or proteins (NMI proteins), extracellular vesicles carrying target genes (NMI genes) or proteins (NMI proteins), and NMI precursor proteins or conjugates or complexes that can be converted into NMI in vivo.

[0069] Preferably, the biological material associated with NMI comprises at least one of f1) to f16): f1) a nucleic acid molecule encoding NMI; f2) an expression cassette comprising the nucleic acid molecule of f1); f3) a vector comprising the nucleic acid molecule of f1); f4) a vector comprising the expression cassette of f2); f5) a transgenic cell line comprising the nucleic acid molecule of f1); f6) a transgenic cell line comprising the expression cassette of f2); f7) a transgenic cell line comprising the vector of f3); f8) a transgenic cell line comprising the vector of f4); f9) a microorganism comprising the nucleic acid molecule of f1); f10) a microorganism comprising the expression cassette of f2); f11) a microorganism comprising the vector of f3); f12) a microorganism comprising the vector of f4); f13) a virus comprising the nucleic acid molecule of f1); f14) a virus comprising the expression cassette of f2); f15) a virus comprising the vector of f3); f16) a virus comprising the vector of f4).

[0070] Preferably, the NMI is a human NMI.

[0071] Preferably, the tumor immunotherapy enhancer further comprises a pharmaceutically acceptable carrier or excipient.

[0072] Preferably, the form of the tumor immunotherapy enhancer makes it suitable for administration by a method selected from the following groups: oral, injection (e.g., direct naked DNA or protein injection, liposome-encapsulated DNA, RNA or protein injection), gold-coated gene gun bombardment, replication-defective bacteria carrying plasmid DNA, replication-defective adenovirus carrying target DNA or target gene-encoded protein, electroporation, intravenous, pulmonary, mucosal, nasal, intraperitoneal, intracranial, intratumoral, sublingual, buccal, and transdermal administration.

[0073] Preferably, the dosage form of the tumor immunotherapy enhancer comprises at least one of capsules, tablets, microcapsules, injections, suppositories, sprays, powders, soft capsules, dripping pills, honey pills, pills, granules, honey pastes, sustained-release preparations, oral liquid preparations, chewable tablets, buccal tablets, transdermal patches, and effervescent tablets.

[0074] Preferably, the tumor immunotherapy enhancer further comprises other substances that improve the efficacy of tumor immunotherapy.

[0075] Preferably, the other substances that improve the efficacy of tumor immunotherapy can be any substances disclosed in the prior art that improve the efficacy of tumor immunotherapy, such as the PARP1 inhibitor disclosed in patent document CN109331021A, the calcium-activated chloride channels (CaCCs) inhibitor disclosed in patent document CN113117088A (such as: T16Ainh-A01 and / or CaCCinh-A01), the animal Bifidobacterium lactis subsp. Lactis disclosed in patent document CN112618577A (such as: Animal Bifidobacterium lactis subsp. NexStrain 02 (Deposit Number: CGMCC No. 20455)), the human intestinal flora whole bacterial composition disclosed in patent document CN111249314A, and the like.

[0076] The third aspect of the present invention provides a tumor immunotherapy enhancer, comprising: (c1) at least one of interferon-induced protein 35kD (IFP35) and interferon-induced protein 35kD (IFP35) promoter; and / or (c2) at least one of N-Myc interacting protein (NMI) and N-Myc interacting protein (NMI) promoter.

[0077] Preferably, the tumor immunotherapy enhancer comprises (c1) and (c2).

[0078] Preferably, the interferon-induced protein 35kD (IFP35) promoter is the interferon-induced protein 35kD (IFP35) promoter in the second aspect of the present invention.

[0079] Preferably, the N-Myc interacting protein (NMI) promoter is the N-Myc interacting protein (NMI) promoter in the second aspect of the present invention.

[0080] Preferably, the tumor immunotherapy enhancer further comprises a pharmaceutically acceptable carrier or excipient.

[0081] Preferably, the form of the tumor immunotherapy enhancer makes it suitable for administration by a method selected from the following groups: oral, injection (e.g., direct naked DNA or protein injection, liposome-encapsulated DNA, RNA or protein injection), gold-coated gene gun bombardment, replication-defective bacteria carrying plasmid DNA, replication-defective adenovirus carrying target DNA or target gene-encoded protein, electroporation, intravenous, pulmonary, mucosal, nasal, intraperitoneal, intracranial, intratumoral, sublingual, buccal, and transdermal administration.

[0082] Preferably, the dosage form of the tumor immunotherapy enhancer comprises at least one of capsules, tablets, microcapsules, injections, suppositories, sprays, powders, soft capsules, dripping pills, honey pills, pills, granules, honey pastes, sustained-release preparations, oral liquid preparations, chewable tablets, buccal tablets, transdermal patches, and effervescent tablets.

[0083] Preferably, the tumor immunotherapy enhancer further comprises other substances that improve the efficacy of tumor immunotherapy.

[0084] Preferably, the other substances that improve the efficacy of tumor immunotherapy can be any substances disclosed in the prior art that improve the efficacy of tumor immunotherapy, such as the PARP1 inhibitor disclosed in patent document CN109331021A, the calcium-activated chloride channels (CaCCs) inhibitor disclosed in patent document CN113117088A (such as: T16Ainh-A01 and / or CaCCinh-A01), the animal Bifidobacterium lactis subsp. Lactis disclosed in patent document CN112618577A (such as: Animal Bifidobacterium lactis subsp. NexStrain 02 (Deposit Number: CGMCC No. 20455)), the human intestinal flora whole bacterial composition disclosed in patent document CN111249314A, and the like.

[0085] The fourth aspect of the present invention provides an anti-tumor pharmaceutical composition comprising: (e1) a tumor immunotherapy synergist; and (e2) a tumor immunotherapy agent;

[0086] The tumor immunotherapy enhancer is the tumor immunotherapy enhancer in the third aspect of the present invention.

[0087] Preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or excipient.

[0088] Preferably, the form of the pharmaceutical composition makes it suitable for administration selected from the group consisting of oral, injection (e.g., direct naked DNA or protein injection, liposome-encapsulated DNA, RNA or protein injection), gold-coated gene gun bombardment, replication-defective bacteria carrying plasmid DNA, replication-defective adenovirus carrying target DNA or target gene-encoded protein, electroporation, intravenous, pulmonary, mucosal, nasal, intraperitoneal, intracranial, intratumoral, sublingual, buccal, and transdermal administration.

[0089] Preferably, the dosage form of the pharmaceutical composition comprises at least one of capsules, tablets, microcapsules, injections, suppositories, sprays, powders, soft capsules, dripping pills, honey pills, pills, granules, honey pastes, sustained-release preparations, oral liquid preparations, chewable tablets, buccal tablets, transdermal patches, and effervescent tablets.

[0090] Preferably, the pharmaceutical composition further comprises instructions.

[0091] Preferably, the instructions describe the number of administrations, dosage and administration method.

[0092] A fifth aspect of the present invention provides an anti-tumor drug kit comprising: (e1) a tumor immunotherapy synergist; and (e2) a tumor immunotherapy agent;

[0093] The tumor immunotherapy synergist is the tumor immunotherapy synergist of the third aspect of the present invention;

[0094] The tumor immunotherapy enhancer and the tumor immunotherapy agent exist independently.

[0095] Preferably, the tumor immunotherapeutic agent further comprises a pharmaceutically acceptable carrier or excipient.

[0096] Preferably, the form of the tumor immunotherapeutic agent makes it suitable for administration by a method selected from the following groups: oral, injection (e.g., direct naked DNA or protein injection, liposome-encapsulated DNA, RNA or protein injection), gold-coated gene gun bombardment, replication-defective bacteria carrying plasmid DNA, replication-defective adenovirus carrying target DNA or target gene-encoded protein, electroporation, intravenous, pulmonary, mucosal, nasal, intraperitoneal, intracranial, intratumoral, sublingual, buccal, and transdermal administration.

[0097] Preferably, the dosage form of the tumor immunotherapy agent comprises at least one of capsules, tablets, microcapsules, injections, suppositories, sprays, powders, soft capsules, dripping pills, honey pills, pills, granules, honey pastes, sustained-release preparations, oral liquid preparations, chewable tablets, buccal tablets, transdermal patches, and effervescent tablets.

[0098] Preferably, the tumor immunotherapy enhancer and the tumor immunotherapy agent are administered simultaneously or in any order, for example, the tumor immunotherapy agent is administered before the tumor immunotherapy enhancer, or the tumor immunotherapy agent is administered after the tumor immunotherapy enhancer, or both are administered simultaneously.

[0099] Preferably, the pharmaceutical kit further comprises instructions.

[0100] Preferably, the instructions describe the number of administrations, dosage and administration method.

[0101] A sixth aspect of the present invention provides a method for detecting the content and / or expression of a marker in a test sample, wherein the marker comprises the IFP35 protein family, wherein the IFP35 protein family comprises interferon-induced protein 35kD (IFP35) and / or N-Myc interacting protein (NMI);

[0102] The method is used for at least one of (1)-(3): (1) evaluation of the efficacy of tumor immunotherapy; (2) prognosis evaluation of tumor immunotherapy; (3) selection of treatment regimen.

[0103] Preferably, tumor immunotherapy efficacy evaluation refers to judging the effect of tumor immunotherapy (whether there is a response to tumor immunotherapy) based on the level of markers.

[0104] Preferably, the prognosis assessment of tumor immunotherapy refers to judging the prognosis (good prognosis or poor prognosis) based on the level of the marker.

[0105] Preferably, the treatment regimen selection refers to selecting whether to use a tumor immunotherapy enhancer (tumor immunotherapy, or tumor immunotherapy combined with a tumor immunotherapy enhancer) based on the level of the marker.

[0106] Preferably, the detection of the content and / or expression of the marker in the test sample includes gene level and / or protein level.

[0107] Preferably, the method for detecting the content and / or expression of the marker in the test sample includes at least one of immunohistochemistry, Western blotting, Northern blotting, PCR, biochip method, nucleic acid sequencing method, and amino acid sequencing method.

[0108] Preferably, the immunohistochemistry method is selected from at least one of immunofluorescence assay, enzyme-linked immunosorbent assay (ELISA) and immunocolloidal gold assay.

[0109] Preferably, the substance for detecting the content and / or expression of the marker in the test sample is selected from one or more of: a substance specific to the marker, a marker-specific probe, a gene chip, and a PCR primer.

[0110] Preferably, the substance specific for the marker comprises any one of (a1) to (a3): (a1) an antibody that specifically binds to the marker; (a2) a ligand protein or polypeptide that specifically binds to the marker; (a3) ​​a non-protein compound that specifically recognizes the marker.

[0111] Preferably, the antibody comprises at least one of a polyclonal antibody, a monoclonal antibody, a single-chain antibody, a functional antibody fragment, an antibody Fab region, a nanobody, a chimeric antibody, and a multispecific antibody.

[0112] Preferably, the test sample is selected from at least one of body fluids, tissues, cells, and excretions of the subject to be tested.

[0113] Preferably, the body fluid comprises at least one of blood and lymph.

[0114] Preferably, the blood comprises at least one of serum, plasma, dried blood spots, and whole blood.

[0115] Preferably, the tissue comprises tumor tissue.

[0116] Preferably, the excrement comprises at least one of urine, feces, and tears.

[0117] Preferably, the subject to be tested includes mammals, such as humans, non-human primates (such as gorillas and apes), rodents (such as rats, mice, and guinea pigs), pets (such as cats and dogs), and livestock (such as horses, cows, sheep, pigs, and rabbits).

[0118] Preferably, the subject to be tested includes humans.

[0119] Preferably, the detected level of the IFP35 protein family is positively correlated with the evaluation of tumor immunotherapy efficacy and tumor immunotherapy prognosis, that is, the higher the level of the detected IFP35 protein family, the better the tumor immunotherapy efficacy and tumor immunotherapy prognosis.

[0120] Preferably, the higher the level of the detected IFP35 protein family, the less the need for combined use of a tumor immunotherapy enhancer.

[0121] Preferably, the markers further include other markers for evaluating the efficacy of tumor immunotherapy, evaluating the prognosis of tumor immunotherapy, and / or selecting treatment regimens.

[0122] Preferably, the other markers for evaluating the efficacy of tumor immunotherapy, evaluating the prognosis of tumor immunotherapy, and / or selecting a treatment regimen can be any marker disclosed in the prior art for evaluating the efficacy of tumor immunotherapy, evaluating the prognosis of tumor immunotherapy, and / or selecting a treatment regimen, such as p62 / SQSTM1 disclosed in patent document CN112014564A, the combination disclosed in patent document CN113358872A (H3Cit, IL-8 and CRP), the combination disclosed in patent document CN110223729A (VPS29, SNRPD3, PSMB2, VCP, RAB7A, PSMB4, C1or43, GAPDH, ACTB), and the combination disclosed in patent document CN109576365A (SCML4, TTC16, KIR2DL3, ART4, LINC00426, C15orf53, HEATR9, KIR2DL1, TRAT1, IL27, SLFN14, KEL, CXCR2P1, KIR3DL2, GBP7, BTN1A1, IFNG, CLEC6A, LINC00494, ITIH1, CLECL1, GHRL, C4orf50, WDR64, PLA2G2D, CR1 L, OR52N4, TTLL6, FCRL6, PIK3CD-AS1, UPK3A, KIF25, LILRA4, RTP5, CLEC4C, KIR2DS4, C9orf139, DBH, FCER2, CCR9, CD40LG, NCR1, CCL 25. RHOXF1, GPR174, TIMD4, IL26, AICDA, KCNJ10, FAM129C, WNT1, FAM180B, KLRC3, CNR2, LINC00158, XKR4, IDO2, FCRL2, FAM92B, ZFR2, PTCRA, DNAJC5B, DNAH8, OCM, GPR25, KLRC4, KIR3DL1, DNASE2B, TCL1A, LOC100240735, IL13, PSMA8, CLEC9A, NME8, BCORP1, NCR2, IL12B , LILRP2, KCNA2, CCKAR, CLNK, FCRL4, SLC18A1, CRYBA4, TNFRSF13B, SSTR3, CR2, LCNL1, TTC24, ITGAD, IL17REL, DAZL, ZNF80, CXorf65,BEND4, FCAMR, CD19, HSF5, FNDC9, TUSC5, ARRDC5, GTSF1L, ATP1A4, CCL1, BFSP2, FCRL1, IL5RA, AIRE, MS4A1, UNQ6494, FAM177B, NPHS1, ASCL4, FAM216B, FRMD1); the combination disclosed in patent document CN110456042A (MIP-1beta, SDF-1alpha, PD-L1, CD28, TIM-3, RANTES and any multiple of IP-10), the levels of other markers for tumor immunotherapy efficacy evaluation, tumor immunotherapy prognosis evaluation, and / or treatment regimen selection detected and tumor immunotherapy efficacy evaluation, tumor immunotherapy prognosis evaluation, and / or treatment regimen selection refer to the above patent documents.

[0123] A seventh aspect of the present invention is to provide a system comprising:

[0124] Device for detecting the content and / or expression of a marker in a test sample: used to detect the content and / or expression of a marker in a test sample;

[0125] Evaluation or selection device: evaluates the efficacy of tumor immunotherapy, assesses the prognosis of tumor immunotherapy, and / or selects a treatment plan based on the content and / or expression of the marker;

[0126] Result output device: outputs tumor immunotherapy efficacy evaluation, tumor immunotherapy prognosis evaluation, and / or treatment plan selection results;

[0127] The markers include the IFP35 protein family, and the IFP35 protein family includes interferon-induced protein 35kD (IFP35) and / or N-Myc interacting protein (NMI).

[0128] Preferably, tumor immunotherapy efficacy evaluation refers to judging the effect of tumor immunotherapy (whether there is a response to tumor immunotherapy) based on the level of markers.

[0129] Preferably, the prognosis assessment of tumor immunotherapy refers to judging the prognosis (good prognosis or poor prognosis) based on the level of the marker.

[0130] Preferably, the treatment regimen selection refers to selecting whether to use a tumor immunotherapy enhancer (tumor immunotherapy, or tumor immunotherapy combined with a tumor immunotherapy enhancer) based on the level of the marker.

[0131] Preferably, the device for detecting the content and / or expression of the marker in the test sample includes a device for detecting the content and / or expression of the marker at the gene level and / or protein level.

[0132] Preferably, the device for detecting the content and / or expression of the marker in the test sample comprises a device for one or more detection techniques or methods selected from the following group: immunohistochemistry, Western blotting, Northern blotting, PCR, biochip method, nucleic acid sequencing method, and amino acid sequencing method.

[0133] Preferably, the immunohistochemistry method is selected from at least one of immunofluorescence assay, enzyme-linked immunosorbent assay (ELISA) and immunocolloidal gold assay.

[0134] Preferably, the test sample of the product is selected from at least one of body fluids, tissues, cells, and excretions of the subject to be tested.

[0135] Preferably, the body fluid comprises at least one of blood and lymph.

[0136] Preferably, the blood comprises at least one of serum, plasma, dried blood spots, and whole blood.

[0137] Preferably, the tissue comprises tumor tissue.

[0138] Preferably, the excrement comprises at least one of urine, feces, and tears.

[0139] Preferably, the subject to be tested includes mammals, such as humans, non-human primates (such as gorillas and apes), rodents (such as rats, mice, and guinea pigs), pets (such as cats and dogs), and livestock (such as horses, cows, sheep, pigs, and rabbits).

[0140] Preferably, the subject to be tested includes humans.

[0141] Preferably, the detected level of the IFP35 protein family is positively correlated with the evaluation of tumor immunotherapy efficacy and tumor immunotherapy prognosis, that is, the higher the level of the detected substance, the better the tumor immunotherapy efficacy and tumor immunotherapy prognosis.

[0142] Preferably, the higher the level of the detected IFP35 protein family, the less the need for combined use of a tumor immunotherapy enhancer.

[0143] Preferably, the markers further include other markers for evaluating the efficacy of tumor immunotherapy, evaluating the prognosis of tumor immunotherapy, and / or selecting treatment regimens.

[0144] Preferably, the other markers for evaluating the efficacy of tumor immunotherapy, evaluating the prognosis of tumor immunotherapy, and / or selecting a treatment regimen can be any marker disclosed in the prior art for evaluating the efficacy of tumor immunotherapy, evaluating the prognosis of tumor immunotherapy, and / or selecting a treatment regimen, such as p62 / SQSTM1 disclosed in patent document CN112014564A, the combination (H3Cit, IL-8 and CRP) disclosed in patent document CN113358872A, the combination (VPS29, SNRPD3, PSMB2, VCP, RAB7A, PSMB4, C1or43, GAPDH, ACTB) disclosed in patent document CN110223729A, and the combination (at least 3 of VPS29, SNRPD3, PSMB2, VCP, RAB7A, PSMB4, C1or43, GAPDH, ACTB) disclosed in patent document CN112014564A. The combination disclosed in CN109576365A (SCML4, TTC16, KIR2DL3, ART4, LINC00426, C15orf53, HEATR9, KIR2DL1, TRAT1, IL27, SLFN14, KEL, CXCR2P1, KIR3DL2, GBP7, BTN1A1, IFNG, CLEC6A, LINC00494, ITIH1, CLECL1, GHRL, C4orf50, WDR64, PLA2G2D, CR1L, OR52N4, TTLL6, FCRL6, PIK3CD-AS1, UPK3A, KIF25, LILRA4, RTP5, CLEC4C, KIR2DS4, C9orf139, DBH, FCER2, CCR9, CD40LG, NCR1, CCL25, RHOXF1, GPR174, TIMD4, IL26, AICDA, KCNJ10, FAM129C, WNT1, FAM180B, KLRC3, CN R2, LINC00158, EC9A, NME8, BCORP1, NCR2, IL12B, LILRP2, KCNA2, CCKAR, CLNK, FCRL4, SLC18A1, CRYBA4, TNFRSF13B, SSTR3, CR2, LCNL1, TTC24, ITGAD, IL17REL, DAZL, ZNF80, CXorf65, BEND4, FCAMR, CD19, HSF5, FNDC9, TUSC5, ARRDC5, GTSF1L, ATP1A4, CCL1, BFSP2, FCRL1, IL5RA, AIRE, MS4A1, UNQ6494, FAM177B, NPHS1,ASCL4, FAM216B, FRMD1); the combination disclosed in patent document CN110456042A (MIP-1beta, SDF-1alpha, PD-L1, CD28, TIM-3, RANTES and any multiple of IP-10), the levels of other markers for tumor immunotherapy efficacy evaluation, tumor immunotherapy prognosis evaluation, and / or treatment regimen selection and tumor immunotherapy efficacy evaluation, tumor immunotherapy prognosis evaluation, and / or treatment regimen selection are referred to the above patent documents.

[0145] The eighth aspect of the present invention is to provide a marker combination comprising the IFP35 protein family, wherein the IFP35 protein family comprises interferon-induced protein 35kD (IFP35) and N-Myc interacting protein (NMI).

[0146] Preferably, the marker combination is used for at least one of (1)-(3): (1) evaluation of the efficacy of tumor immunotherapy; (2) evaluation of the prognosis of tumor immunotherapy; (3) selection of treatment regimen.

[0147] Preferably, tumor immunotherapy efficacy evaluation refers to judging the effect of tumor immunotherapy (whether there is a response to tumor immunotherapy) based on the level of markers.

[0148] Preferably, the prognosis assessment of tumor immunotherapy refers to judging the prognosis (good prognosis or poor prognosis) based on the level of the marker.

[0149] Preferably, the treatment regimen selection refers to selecting whether to use a tumor immunotherapy enhancer (tumor immunotherapy, or tumor immunotherapy combined with a tumor immunotherapy enhancer) based on the level of the marker.

[0150] Preferably, the marker combination is derived from at least one of body fluids, tissues, cells, and excretions of the subject to be tested.

[0151] Preferably, the body fluid comprises at least one of blood and lymph.

[0152] Preferably, the blood comprises at least one of serum, plasma, dried blood spots, and whole blood.

[0153] Preferably, the tissue comprises tumor tissue.

[0154] Preferably, the excrement comprises at least one of urine, feces, and tears.

[0155] Preferably, the subject to be tested includes mammals, such as humans, non-human primates (such as gorillas and apes), rodents (such as rats, mice, and guinea pigs), pets (such as cats and dogs), and livestock (such as horses, cows, sheep, pigs, and rabbits).

[0156] Preferably, the subject to be tested includes humans.

[0157] Preferably, the detected level of the IFP35 protein family is positively correlated with the evaluation of tumor immunotherapy efficacy and tumor immunotherapy prognosis, that is, the higher the level of the detected IFP35 protein family, the better the tumor immunotherapy efficacy and tumor immunotherapy prognosis.

[0158] Preferably, the higher the level of the detected IFP35 protein family, the less the need for combined use of a tumor immunotherapy enhancer.

[0159] Preferably, the markers further include other markers for evaluating the efficacy of tumor immunotherapy, evaluating the prognosis of tumor immunotherapy, and / or selecting treatment regimens.

[0160] Preferably, the other markers for evaluating the efficacy of tumor immunotherapy, evaluating the prognosis of tumor immunotherapy, and / or selecting a treatment regimen can be any marker disclosed in the prior art for evaluating the efficacy of tumor immunotherapy, evaluating the prognosis of tumor immunotherapy, and / or selecting a treatment regimen, such as p62 / SQSTM1 disclosed in patent document CN112014564A, the combination (H3Cit, IL-8 and CRP) disclosed in patent document CN113358872A, the combination (VPS29, SNRPD3, PSMB2, VCP, RAB7A, PSMB4, C1or43, GAPDH, ACTB) disclosed in patent document CN110223729A, and the combination (at least 3 of VPS29, SNRPD3, PSMB2, VCP, RAB7A, PSMB4, C1or43, GAPDH, ACTB) disclosed in patent document CN112014564A. The combination disclosed in CN109576365A (SCML4, TTC16, KIR2DL3, ART4, LINC00426, C15orf53, HEATR9, KIR2DL1, TRAT1, IL27, SLFN14, KEL, CXCR2P1, KIR3DL2, GBP7, BTN1A1, IFNG, CLEC6A, LINC00494, ITIH1, CLECL1, GHRL, C4orf50, WDR64, PLA2G2D, CR1L, OR52N4, TTLL6, FCRL6, PIK3CD-AS1, UPK3A, KIF25, LILRA4, RTP5, CLEC4C, KIR2DS4, C9orf139, DBH, FCER2, CCR9, CD40LG, NCR1, CCL25, RHOXF1, GPR174, TIMD4, IL26, AICDA, KCNJ10, FAM129C, WNT1, FAM180B, KLRC3, CN R2, LINC00158, EC9A, NME8, BCORP1, NCR2, IL12B, LILRP2, KCNA2, CCKAR, CLNK, FCRL4, SLC18A1, CRYBA4, TNFRSF13B, SSTR3, CR2, LCNL1, TTC24, ITGAD, IL17REL, DAZL, ZNF80, CXorf65, BEND4, FCAMR, CD19, HSF5, FNDC9, TUSC5, ARRDC5, GTSF1L, ATP1A4, CCL1, BFSP2, FCRL1, IL5RA, AIRE, MS4A1, UNQ6494, FAM177B, NPHS1,ASCL4, FAM216B, FRMD1); the combination disclosed in patent document CN110456042A (MIP-1beta, SDF-1alpha, PD-L1, CD28, TIM-3, RANTES and any multiple of IP-10), the levels of other markers for tumor immunotherapy efficacy evaluation, tumor immunotherapy prognosis evaluation, and / or treatment regimen selection and tumor immunotherapy efficacy evaluation, tumor immunotherapy prognosis evaluation, and / or treatment regimen selection are referred to the above patent documents.

[0161] The ninth aspect of the present invention is to provide a product comprising substances for detecting the marker combination according to the seventh aspect of the present invention.

[0162] Preferably, the product is used for at least one of (1)-(3): (1) evaluation of the efficacy of tumor immunotherapy; (2) prognosis evaluation of tumor immunotherapy; (3) selection of treatment regimen.

[0163] Preferably, tumor immunotherapy efficacy evaluation refers to judging the effect of tumor immunotherapy (whether there is a response to tumor immunotherapy) based on the level of markers.

[0164] Preferably, the prognosis assessment of tumor immunotherapy refers to judging the prognosis (good prognosis or poor prognosis) based on the level of the marker.

[0165] Preferably, the treatment regimen selection refers to selecting whether to use a tumor immunotherapy enhancer (tumor immunotherapy, or tumor immunotherapy combined with a tumor immunotherapy enhancer) based on the level of the marker.

[0166] Preferably, the substance for detecting the marker combination comprises a substance for quantitatively detecting the marker combination.

[0167] Preferably, the substance for detecting the marker combination comprises a substance for detecting the marker combination at the gene level and / or protein level.

[0168] Preferably, the substance comprises a substance for one or more detection techniques or methods selected from the group consisting of immunohistochemistry, Western blotting, Northern blotting, PCR, biochip, nucleic acid sequencing, and amino acid sequencing.

[0169] Preferably, the immunohistochemistry method is selected from at least one of immunofluorescence assay, enzyme-linked immunosorbent assay (ELISA) and immunocolloidal gold assay.

[0170] Preferably, the substance for detecting the marker is selected from one or more of: a substance specific to the marker combination, a probe specific to the marker combination, a gene chip, and a PCR primer.

[0171] Preferably, the substance specific for the marker combination comprises any one of (a1) to (a3): (a1) an antibody that specifically binds to the marker combination; (a2) a ligand protein or polypeptide that specifically binds to the marker combination; (a3) ​​a non-protein compound that specifically recognizes the marker combination.

[0172] Preferably, the antibody comprises at least one of a polyclonal antibody, a monoclonal antibody, a single-chain antibody, a functional antibody fragment, an antibody Fab region, a nanobody, a chimeric antibody, and a multispecific antibody.

[0173] Preferably, the product comprises at least one of a reagent, a kit, a test paper, a chip, and a system.

[0174] Preferably, the test sample of the product is selected from at least one of body fluids, tissues, cells, and excretions of the subject to be tested.

[0175] Preferably, the body fluid comprises at least one of blood and lymph.

[0176] Preferably, the blood comprises at least one of serum, plasma, dried blood spots, and whole blood.

[0177] Preferably, the tissue comprises tumor tissue.

[0178] Preferably, the excrement comprises at least one of urine, feces, and tears.

[0179] Preferably, the subject to be tested includes mammals, such as humans, non-human primates (such as gorillas and apes), rodents (such as rats, mice, and guinea pigs), pets (such as cats and dogs), and livestock (such as horses, cows, sheep, pigs, and rabbits).

[0180] Preferably, the subject to be tested includes humans.

[0181] The tenth aspect of the present invention provides the use of a marker as a target in the development of products related to tumor immunotherapy, wherein the marker is the marker in the first aspect of the present invention.

[0182] Preferably, the product is:

[0183] Reagents, kits, test strips, chips, or systems; or

[0184] tumor immunotherapeutic agents;

[0185] The reagent, kit, test paper, chip, or system is used for at least one of (1)-(3): (1) evaluation of the efficacy of tumor immunotherapy; (2) prognosis evaluation of tumor immunotherapy; (3) selection of treatment regimen.

[0186] Preferably, tumor immunotherapy efficacy evaluation refers to judging the effect of tumor immunotherapy (whether there is a response to tumor immunotherapy) based on the level of markers.

[0187] Preferably, the prognosis assessment of tumor immunotherapy refers to judging the prognosis (good prognosis or poor prognosis) based on the level of the marker.

[0188] Preferably, the treatment regimen selection refers to selecting whether to use a tumor immunotherapy enhancer (tumor immunotherapy, or tumor immunotherapy combined with a tumor immunotherapy enhancer) based on the level of the marker.

[0189] The eleventh aspect of the present invention provides a tumor immunotherapy synergist and a tumor immunotherapy agent for use in preparing a drug for treating tumors. The tumor immunotherapy synergist is the tumor immunotherapy synergist of the third aspect of the present invention.

[0190] The twelfth aspect of the present invention provides a method for improving the efficacy of tumor immunotherapy, which increases the content and / or activity of the IFP35 protein family in an individual undergoing tumor immunotherapy, wherein the IFP35 protein family includes interferon-induced protein 35kD (IFP35) and / or N-Myc interacting protein (NMI).

[0191] Preferably, the method for increasing the content and / or activity of the IFP35 protein family in an individual undergoing tumor immunotherapy is to administer the tumor immunotherapy enhancer according to the third aspect of the present invention to the individual.

[0192] Preferably, the subject comprises a mammal, such as a human, non-human primate (e.g., gorilla, ape), rodent (e.g., rat, mouse, guinea pig), pet (e.g., cat, dog), livestock (e.g., horse, cow, sheep, pig, rabbit).

[0193] Preferably, the individual comprises a human.

[0194] The thirteenth aspect of the present invention provides a method for treating tumors, comprising administering to an individual an effective amount of the anti-tumor pharmaceutical composition of the fourth aspect of the present invention and / or the anti-tumor drug set of the fifth aspect of the present invention.

[0195] Preferably, the subject comprises a mammal, such as a human, non-human primate (e.g., gorilla, ape), rodent (e.g., rat, mouse, guinea pig), pet (e.g., cat, dog), livestock (e.g., horse, cow, sheep, pig, rabbit).

[0196] Preferably, the individual comprises a human.

[0197] The fourteenth aspect of the present invention provides a method for constructing a model with poor tumor immunotherapy efficacy, which reduces the content and / or activity of the IFP35 protein family in the model, wherein the IFP35 protein family includes interferon-induced protein 35kD (IFP35) and / or N-Myc interacting protein (NMI).

[0198] Preferably, the method for reducing the content and / or activity of the IFP35 protein family in the model is to knock out the gene of the IFP35 protein family.

[0199] Preferably, the model comprises an animal model.

[0200] Preferably, the animal comprises at least one of rats and mice.

[0201] Preferably, the poor efficacy of tumor immunotherapy includes no response to tumor immunotherapy.

[0202] The fifteenth aspect of the present invention provides a method for constructing a model with good tumor immunotherapy efficacy, which increases the content and / or activity of the IFP35 protein family in the model, wherein the IFP35 protein family includes interferon-induced protein 35kD (IFP35) and / or N-Myc interacting protein (NMI).

[0203] Preferably, the method for increasing the content and / or activity of the IFP35 protein family in the model is to administer an effective amount of the tumor immunotherapy enhancer according to the third aspect of the present invention to the model.

[0204] Preferably, the model comprises an animal model.

[0205] Preferably, the animal comprises at least one of rats and mice.

[0206] Preferably, the poor efficacy of tumor immunotherapy includes response to tumor immunotherapy.

[0207] Among the above aspects,

[0208] Preferably, the tumor comprises a solid tumor and / or a hematological tumor.

[0209] Preferably, the solid tumor comprises head and neck squamous cell carcinoma, liver cancer, colorectal cancer, bladder cancer, breast cancer, cervical cancer, prostate cancer, glioma, melanoma, pancreatic cancer, nasopharyngeal cancer, lung cancer, gastric cancer, adrenal cortical carcinoma, pararenal cortical carcinoma, anal cancer, appendix cancer, astrocytoma, atypical teratoma, rhabdoid tumor, basal cell carcinoma, bile duct cancer, Bladder cancer, bone cancer, brain tumor, bronchial tumor, Burkitt lymphoma, carcinoid tumor, heart tumor, bile duct carcinoma, chordoma, colorectal cancer, craniopharyngioma, ductal carcinoma in situ, embryonal tumor, endometrial cancer, ependymoma, esophageal cancer, olfactory neuroblastoma, intracranial germ cell tumor, extragonadal germ cell tumor, eye cancer, fallopian tube cancer, gallbladder cancer, head and neck cancer, hypopharyngeal cancer, Kaposi's sarcoma, kidney cancer, Langerhans cell histiocytosis, laryngeal cancer, lip cancer, oral cancer, Merkel cell carcinoma, malignant mesothelioma, multiple endocrine neoplasia syndrome, mycosis fungoides, nasal cavity and paranasal cancer, nerve At least one of: blastoma, non-small cell lung cancer, ovarian cancer, pancreatic neuroendocrine tumor, islet cell tumor, papillomatosis, paraganglioma, sinonasal cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pituitary tumor, pleuropulmonary blastoma, primary peritoneal cancer, retinoblastoma, salivary gland tumor, sarcoma, Sézale syndrome, skin cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, squamous cell carcinoma, testicular cancer, thymoma and thymic cancer, thyroid cancer, urethral cancer, uterine cancer, endometrial and uterine sarcoma, vaginal cancer, vascular tumor, vulvar cancer and single myeloma.

[0210] Preferably, the hematological tumor is selected from at least one of B-cell acute lymphoid leukemia, T-cell acute lymphoid leukemia, acute lymphoid leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia, B-cell prolymphocytic leukemia, blastic plasmacytoid dendritic cell neoplasm, Burkitt's lymphoma, diffuse large B-cell lymphoma, follicular lymphoma, hairy cell leukemia, small cell or large cell-follicular lymphoma, MALT lymphoma, mantle cell lymphoma, marginal zone lymphoma, multiple myeloma, non-Hodgkin's lymphoma, plasmablastic lymphoma, plasmacytoid dendritic cell neoplasm, Waldenstrom macroglobulinemia and preleukemia.

[0211] Preferably, the immunotherapy comprises at least one of monoclonal antibody therapy, immune checkpoint inhibitor therapy, adoptive cell therapy, oncolytic virus therapy, and tumor vaccine therapy.

[0212] Preferably, the target of the monoclonal antibody is selected from one of CD20, HER2, VEGF / VEGFR, EGFR, CD19, FGL1, CD47, CD3, CD30, CD33, CD38, CD52, αVβ3, α5β1, FAP, Tenascin, CEA, EPCAM, PSMA, GAN-GD2, GAN-GD3, GM2, and IGF-IR.

[0213] Preferably, the immune checkpoint inhibitor is an inhibitor that acts on T cell negative co-stimulatory (co-inhibitory) molecules and / or their respective ligands.

[0214] Preferably, the negative co-stimulatory (co-inhibitory) molecules acting on T cells and / or their respective ligands are selected from one or more of CTLA-4, PD-1, PD-L1, PD-L2, B7-1, B7-2, B7-H3, B7-H4, B7-H6, A2aR, IDO, TIM-3, BTLA, VISTA, TIGIT, LAG-3, CD40, CD20, CD96, CD73, CD160, STING, CEA, CD47, PVRIG, LAIR1, 2B4, KIR, CEACAM1, GARP, PS, CSF1R, CD94 / NKG2A, TDO, TNFR and DcR3.

[0215] Preferably, the inhibitors acting on T cell negative co-stimulatory (co-inhibitory) molecules and / or their respective ligands comprise any one of (b1) to (b3): ​​(b1) antibodies that specifically bind (neutralize) T cell negative co-stimulatory (co-inhibitory) molecules and / or their respective ligands; (b2) ligand proteins or polypeptides that specifically bind (neutralize) T cell negative co-stimulatory (co-inhibitory) molecules and / or their respective ligands; (b3) non-protein compounds that specifically bind (neutralize) T cell negative co-stimulatory (co-inhibitory) molecules and / or their respective ligands.

[0216] Preferably, the inhibitor of the ligand of the T cell negative co-stimulatory (co-inhibitory) molecule is selected from: CTLA-4 antibodies (such as ipilimumab, ticilimumab (CP-675,206), AGEN-1884, ATOR-1015, MGD019 (PD-1 / CTLA-4 dual antibody)), PD-1 antibodies (such as nivolumab, pembrolizumab, tremelimu mab), tislelizumab (BGB-A317), spartalizumab, MEDI0680, PDR001, FAZ053, MGA012 (retifanlimab), sintilimab, toripalimab, cemiplimab, MGD019 (PD-1 / CTLA-4 dual antibody), MGD013 (tebotelimab,PD-1 / LAG-3 dual antibodies), PD-L1 antibodies (e.g., atezolizumab, camrelizumab, durvalumab, avelumab, LY3300054, CX-072 (Proclaim-CX-072), FAZ053, KN035, MDX-1105), PD-L2 antibodies, B7-1 antibodies, B7-2 antibodies, B7-H3 antibodies (e.g., enoblituzumab), mAb), MGD009, MGC018), B7-H4 antibody, B7-H6 antibody, A2aR antibody (CPI-444, PBF509), IDO antibody (e.g., GDC0919 (navoximod), epacadostat, indoximid, BMS986205), TIM-3 antibody (e.g., TSR022 (TIM-3 monoclonal antibody), MBG453 (TIM-3 monoclonal antibody)), BTLA antibody, VISTA antibody, TIGIT antibody (e.g., BMS-986207, AB 154, COM902 (CGEN-15137), OMP-313M32), LAG-3 antibodies (e.g., BMS 986016, MK-4280 (28G-10), REGN3767, GSK2831781, IMP731 (H5L7BW), BAP050, IMP-701 (LAG-5250), IMP321, TSR-033, LAG525, BI 754111, FS-118, MGD013 (tebotelimab, PD-1 / LAG-3 dual antibody)), CD40 antibodies (e.g., BMS3h-56, lucatumumab (HCD122 and CHIR-12.12), CHIR-5.9, or dacetuzumab (huS2C6, PRO 64553, RG3636, SGN 14, SGN-40)), CD20 antibodies (e.g., rituximab (RITUXAN; IDEC-102; IDEC-C2B8), ABP 798, ofatumumab or obinutuzumab), CD96 antibodies, CD73 antibodies (e.g., MEDI9447 (oleclumab)), CD160 antibodies (e.g., BY55), STING antibodies, CEA antibodies (e.g., cergutuzumab amunaleukin (RG7813,RO-6895882) or RG7802 (RO6958688)), CD47 antibodies (such as HuF9-G4, CC-90002, TTI-621, ALX148, NI-1701, NI-1801, SRF231 or Effi-DEM), PVRIG antibodies (such as COM701 (CGEN-15029)), LAIR1 antibodies, 2B4 antibodies, KIR antibodies (such as lirilumab (1-7F9, B MS-986015, IPH2101), IPH4102), CEACAM1 antibodies (such as CM-24 (MK-6018)), GARP antibodies (such as ARGX-115), PS antibodies, CSF1R antibodies (such as pexidartinib, LY3022855, FPA008, BLZ945), CD94 / NKG2A antibodies, TDO antibodies, TNFR antibodies and DcR3 antibodies.

[0217] Preferably, the adoptive cell therapy comprises at least one of: chimeric antigen receptor T cell (CAR-T) therapy, chimeric antigen receptor NK cell (CAR-NK) therapy, T cell receptor chimeric T cell (TCR-T) therapy, tumor infiltrating immune cell (TILs) therapy, cytokine-induced killer (CIK) cell therapy, lymphokine-activated killer (LAK) cell therapy, and natural killer (NK) cell therapy.

[0218] Preferably, the oncolytic virus comprises at least one of alphavirus, adenovirus, vaccinia virus, Sindbis virus, Seneca Valley virus, coxsackie virus, measles virus, reovirus, vaccinia virus, Newcastle disease virus, vesicular stomatitis virus, herpes simplex virus, poliovirus, influenza virus, mumps virus and parvovirus; further comprises at least one of alphavirus, adenovirus, vaccinia virus, measles virus, vesicular stomatitis virus, and herpes simplex virus.

[0219] Preferably, the alphavirus comprises at least one of M1 virus and Geta virus.

[0220] Preferably, the tumor vaccine comprises at least one of a dendritic cell (DCs) vaccine, a nucleic acid vaccine, and a polypeptide vaccine.

[0221] Preferably, the tumor immunotherapy agent comprises at least one of: a monoclonal antibody, an immune checkpoint inhibitor, an immune cell, an oncolytic virus, and a tumor vaccine.

[0222] Preferably, the target of the monoclonal antibody is selected from one of CD20, HER2, VEGF / VEGFR, EGFR, CD19, FGL1, CD47, CD3, CD30, CD33, CD38, CD52, αVβ3, α5β1, FAP, Tenascin, CEA, EPCAM, PSMA, GAN-GD2, GAN-GD3, GM2, and IGF-IR.

[0223] Preferably, the immune checkpoint inhibitor is an inhibitor that acts on T cell negative co-stimulatory (co-inhibitory) molecules and / or their respective ligands.

[0224] Preferably, the negative co-stimulatory (co-inhibitory) molecules acting on T cells and / or their respective ligands are selected from one or more of CTLA-4, PD-1, PD-L1, PD-L2, B7-1, B7-2, B7-H3, B7-H4, B7-H6, A2aR, IDO, TIM-3, BTLA, VISTA, TIGIT, LAG-3, CD40, CD20, CD96, CD73, CD160, STING, CEA, CD47, PVRIG, LAIR1, 2B4, KIR, CEACAM1, GARP, PS, CSF1R, CD94 / NKG2A, TDO, TNFR and DcR3.

[0225] Preferably, the inhibitors acting on T cell negative co-stimulatory (co-inhibitory) molecules and / or their respective ligands comprise any one of (b1) to (b3): ​​(b1) antibodies that specifically bind (neutralize) T cell negative co-stimulatory (co-inhibitory) molecules and / or their respective ligands; (b2) ligand proteins or polypeptides that specifically bind (neutralize) T cell negative co-stimulatory (co-inhibitory) molecules and / or their respective ligands; (b3) non-protein compounds that specifically bind (neutralize) T cell negative co-stimulatory (co-inhibitory) molecules and / or their respective ligands.

[0226] Preferably, the inhibitor of the ligand of the T cell negative co-stimulatory (co-inhibitory) molecule is selected from: CTLA-4 antibodies (such as ipilimumab, ticilimumab (CP-675,206), AGEN-1884, ATOR-1015, MGD019 (PD-1 / CTLA-4 dual antibody)), PD-1 antibodies (such as nivolumab, pembrolizumab, tremelimu mab), tislelizumab (BGB-A317), spartalizumab, MEDI0680, PDR001, FAZ053, MGA012 (retifanlimab), sintilimab, toripalimab, cemiplimab, MGD019 (PD-1 / CTLA-4 dual antibody), MGD013 (tebotelimab,PD-1 / LAG-3 dual antibodies), PD-L1 antibodies (e.g., atezolizumab, camrelizumab, durvalumab, avelumab, LY3300054, CX-072 (Proclaim-CX-072), FAZ053, KN035, MDX-1105), PD-L2 antibodies, B7-1 antibodies, B7-2 antibodies, B7-H3 antibodies (e.g., enoblituzumab), mAb), MGD009, MGC018), B7-H4 antibody, B7-H6 antibody, A2aR antibody (CPI-444, PBF509), IDO antibody (e.g., GDC0919 (navoximod), epacadostat, indoximid, BMS986205), TIM-3 antibody (e.g., TSR022 (TIM-3 monoclonal antibody), MBG453 (TIM-3 monoclonal antibody)), BTLA antibody, VISTA antibody, TIGIT antibody (e.g., BMS-986207, AB 154, COM902 (CGEN-15137), OMP-313M32), LAG-3 antibodies (e.g., BMS 986016, MK-4280 (28G-10), REGN3767, GSK2831781, IMP731 (H5L7BW), BAP050, IMP-701 (LAG-5250), IMP321, TSR-033, LAG525, BI 754111, FS-118, MGD013 (tebotelimab, PD-1 / LAG-3 dual antibody)), CD40 antibodies (e.g., BMS3h-56, lucatumumab (HCD122 and CHIR-12.12), CHIR-5.9, or dacetuzumab (huS2C6, PRO 64553, RG3636, SGN 14, SGN-40)), CD20 antibodies (e.g., rituximab (RITUXAN; IDEC-102; IDEC-C2B8), ABP 798, ofatumumab or obinutuzumab), CD96 antibodies, CD73 antibodies (e.g., MEDI9447 (oleclumab)), CD160 antibodies (e.g., BY55), STING antibodies, CEA antibodies (e.g., cergutuzumab amunaleukin (RG7813,RO-6895882) or RG7802 (RO6958688)), CD47 antibodies (e.g., HuF9-G4, CC-90002, TTI-621, ALX148, NI-1701, NI-1801, SRF231, or Effi-DEM), PVRIG antibodies (e.g., COM701 (CGEN-15029)), LAIR1 antibodies, 2B4 antibodies, KIR antibodies (e.g., lirilumab (1-7F9, BMS-986015, IPH2101), IPH4102), CEACAM1 antibodies (e.g., CM-24 (MK-6018)), GARP One or more of antibodies (such as ARGX-115), PS antibodies, CSF1R antibodies (such as pexidartinib, LY3022855, FPA008, BLZ945), CD94 / NKG2A antibodies, TDO antibodies, TNFR antibodies, and DcR3 antibodies.

[0227] Preferably, the immune cells include: chimeric antigen receptor T cells (CAR-T), chimeric antigen receptor NK cells (CAR-NK), T cell receptor chimeric T cells (TCR-T), tumor infiltrating immune cells (TILs), cytokine-induced killer (CIK) cells, lymphokine-activated killer (LAK) cells, and natural killer (NK) cells.

[0228] Preferably, the oncolytic virus comprises at least one of alphavirus, adenovirus, vaccinia virus, Sindbis virus, Seneca Valley virus, coxsackie virus, measles virus, reovirus, vaccinia virus, Newcastle disease virus, vesicular stomatitis virus, herpes simplex virus, poliovirus, influenza virus, mumps virus and parvovirus; further comprises at least one of alphavirus, adenovirus, vaccinia virus, measles virus, vesicular stomatitis virus, and herpes simplex virus.

[0229] Preferably, the alphavirus comprises at least one of M1 virus and Geta virus.

[0230] Preferably, the tumor vaccine comprises at least one of a dendritic cell (DCs) vaccine, a nucleic acid vaccine, and a polypeptide vaccine.

[0231] The beneficial effects of the present invention are:

[0232] The present invention discloses for the first time that the IFP35 protein family (interferon-induced protein 35kD (IFP35) and / or N-Myc interacting protein (NMI)) can be used as markers for evaluating the efficacy of tumor immunotherapy, evaluating the prognosis of tumor immunotherapy, and / or selecting treatment regimens; the efficacy of tumor immunotherapy can be improved by using promoters of the IFP35 protein family (interferon-induced protein 35kD (IFP35) and / or N-Myc interacting protein (NMI)) and / or IFP35 protein family (interferon-induced protein 35kD (IFP35) and / or N-Myc interacting protein (NMI)). BRIEF DESCRIPTION OF THE DRAWINGS

[0233] Figure 1 is a comparison of NMI levels in 17 serum samples from HNSCC patients.

[0234] FIG2 is a graph showing the results of ROC curve analysis of the performance of NMI in determining the pathological response of HNSCC patients. DETAILED DESCRIPTION

[0235] The present invention is further described in detail below through specific examples.

[0236] Unless otherwise specified, the materials and reagents used in this example were obtained from commercial sources.

[0237] Example 1 Application of NMI and / or IFP35 in the Evaluation of Tumor Immunotherapy Efficacy

[0238] 1. Application of NMI in evaluating the efficacy of immunotherapy for head and neck squamous cell carcinoma (HNSCC)

[0239] 1. Clinical needs:

[0240] Pathological response means that the patient receives immunotherapy before surgery, and no tumor cells are found in the postoperative tissue specimen pathological examination, which means that the preoperative immunotherapy is effective and the tumor cells are killed.

[0241] Head and neck tumors are one of the common malignant tumors, and the most common pathological type is squamous cell carcinoma. In my country, the incidence of head and neck squamous cell carcinoma (HNSCC) ranks sixth and the mortality rate ranks seventh. After preoperative immunological treatment of patients with head and neck squamous cell carcinoma (HNSCC) (viral / non-viral related), some patients will have a pathological response, while some patients will not. To determine whether there is a pathological response or not, it is necessary to undergo surgery to obtain tissue for pathological testing. However, this is quite difficult in actual clinical practice (such as patients who discharge themselves from the hospital after immunotherapy or do not agree to undergo surgery). Therefore, finding a marker that can quickly determine (non-surgical tissue sampling for pathological testing) whether a patient has a pathological response after immunological treatment is a major need in the clinical diagnosis and treatment of HNSCC.

[0242] 2. Clinical samples:

[0243] Serum samples of HNSCC patients were provided by several hospitals including the Nanfang Hospital of Southern Medical University.

[0244] Patient inclusion criteria: 1) patients with pathological diagnosis of HNSCC; 2) p16 test negative; 3) 2-4 courses of immunotherapy before surgery; 4) pathological examination after surgery.

[0245] Immunotherapy options include: 1) pembrolizumab / Keytruda; 2) nivolumab / Opdivo; 3) tilelizumab / tislelizumab; 4) toripalimab / Toyi; 5) sintilimab / Tyvyt; 6) carrelizumab / Erica; 7) cetuximab.

[0246] Sample collection: Blood was collected after each course of immunization, serum was separated, and stored at -80℃ for future use.

[0247] 3. Detection of NMI in serum samples

[0248] Serum samples were tested for NMI content using a magnetic microparticle chemiluminescence immunoassay and a fully automatic chemiluminescence immunoassay instrument (Guangzhou Darui Biotechnology Co., Ltd.). The test steps are as follows:

[0249] (1) Antibody-coated magnetic beads:

[0250] 1) Activation of Magnetic Microspheres: Pipette 20 mg of carboxyl-modified magnetic microspheres with a diameter of approximately 1 μm into a 2 mL centrifuge tube. Separate the magnetic microspheres from the liquid using a magnetic stand. Wash the magnetic microspheres four times with 1 mL of 0.1 M sodium 2-(N-morpholinoline)ethanesulfonate (MES), pH 6.0. Add 1 mL of 0.1 M MES, pH 6.0, 10 μL of 20 mg / mL 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, and 50 μL of 20 mg / mL N-hydroxysuccinimide to the washed magnetic microspheres, and shake at room temperature for 1 hour.

[0251] 2) Antibody-Magnetic Bead Coupling: The activated magnetic microspheres were placed on a magnetic rack to separate the magnetic microspheres from the liquid. The magnetic microspheres were washed four times with 1 mL of 0.1 M 2-(N-morpholino)ethanesulfonic acid sodium salt (MES) at pH 6.0. 0.5 mg of the coated antibody was added, and 0.1 M 2-(N-morpholino)ethanesulfonic acid sodium salt (MES) at pH 6.0 was added to bring the volume of the magnetic microsphere suspension containing the antibody to 1 mL. The suspension was incubated at room temperature with continuous rotation for 15 hours.

[0252] 3) Blocking: 1.5% (v / v) ethanolamine was added to the antibody-conjugated magnetic microsphere suspension, and the mixture was incubated at room temperature with continuous rotation for 30 minutes. The magnetic microsphere suspension was then placed on a magnetic stand for magnetic separation, washed four times with 1 mL of 0.1 M 2-(N-morpholino)ethanesulfonic acid sodium salt (MES), pH 6.0, and incubated at room temperature with continuous rotation for 2 hours with 1 mL of 0.05 M Tris-HCl buffer (pH 7.5) containing 2% (w / v) BSA and 1% (w / v) trehalose. The mixture was then placed on a magnetic stand for magnetic separation, and washed four times with 1 mL of 0.05 M Tris-HCl buffer (pH 7.5) containing 2% (w / v) BSA and 1% (w / v) trehalose. Finally, 1 mL of 0.05 M Tris-HCl buffer (pH 7.5) containing 2% (w / v) BSA and 1% (w / v) trehalose was added to resuspend the magnetic beads to obtain magnetic microspheres coated with monoclonal antibody 98G10 (for details of monoclonal antibody 98G10, see WO2024213001A1); the above-mentioned magnetic microspheres coated with the coating antibody were diluted with 0.05 M Tris-HCl buffer (pH 7.5) containing 2% (w / v) BSA and 1% (w / v) trehalose at a ratio of 1:100, aliquoted, and stored in a medical refrigerator at 2-8°C for future use.

[0253] (2) Acridinium ester labeled antibodies:

[0254] 1) Antibody buffer exchange: 1.0 mg of labeled antibody was added to an ultrafiltration centrifuge tube with a cutoff of 50 kDa and centrifuged at 8000 rpm at 4°C for 5 minutes for concentration; the waste liquid was discarded, and PB buffer containing 0.02 M disodium hydrogen phosphate, 0.002 M potassium dihydrogen phosphate, pH 6.5 was added to the ultrafiltration centrifuge tube to make the volume in the ultrafiltration centrifuge tube approximately 400 μL; centrifuged at 8000 rpm at 4°C for 5 minutes, the waste liquid was discarded, and this operation was repeated 4 times; the ultrafiltration centrifuge tube was inverted and centrifuged at 3000 rpm at 4°C for 1 minute to collect the antibody solution; 100 μL of PB buffer was added to the ultrafiltration centrifuge tube, and after standing for 5 minutes, the ultrafiltration centrifuge tube was inverted again and centrifuged at 3000 rpm at 4°C for 1 minute to collect the solution, and the two collected antibody solutions were combined;

[0255] 2) Acridinium ester-antibody conjugation: Add 0.05 mg of acridinium salt to the above antibody and incubate at 26°C with shaking in the dark for 19 ± 1 h;

[0256] 3) Purification: Acridinium ester-labeled monoclonal antibody B-8 (for details of monoclonal antibody B-8, see WO2024213001A1) was purified using a Sephadex™ G50 gel column coupled with a high-efficiency protein purification system. The eluent was the aforementioned pH 6.5 PB buffer. The cleanliness of the gel chromatography column was checked using the UV detection function of the purification system. Purification was initiated after all baselines stabilized, and the solution of the first protein peak was collected as the target antibody solution.

[0257] 4) Storage: The purified and collected acridinium ester-labeled antibody was filtered through a 0.22 μm syringe filter, and its protein concentration was determined using a Nanodrop. Finally, 1% of the total volume of a protein stabilizer containing 10% (v / v) glycerol and 0.1% (v / v) Procline 300 was added. The acridinium salt-labeled monoclonal antibody B-8 was diluted 1:400 with 0.05 M Tris-HCl buffer containing 1% (w / v) trehalose, 0.1% (v / v) Tween-20, 1 mM disodium ethylenediaminetetraacetic acid, pH 7.2. The product was aliquoted and stored in a medical refrigerator at 2-8°C.

[0258] (3) hNMI calibrator: lyophilized powder, prepared by diluting recombinant human hNMI protein to 0.00 pg / mL, 200.00 pg / mL, and 2000.00 pg / mL in 0.05 M Tris-HCl buffer containing 5% (w / v) BSA, 1% (w / v) trehalose, 0.1% (v / v) Tween-20, and 0.1% (v / v) Proclin 300, pH 7.8, and lyophilizing in 1 mL / vial aliquots.

[0259] (4) Reaction buffer: 0.1% (v / v) Tween-20, 0.1% (v / v) Proclin 300, 0.025 M Tris-HCl buffer, pH 7.2

[0260] (5) Clinical sample testing: The NMI content in clinical samples was tested using the fully automatic chemiluminescence detector DR-CL2000. The above-prepared components and clinical samples were placed into the instrument according to the instrument operating instructions, and then the instrument was started to automatically detect the samples.

[0261] 4. Detection of IFP35 in Serum Samples

[0262] The IFP35 detection kit (human interferon-induced protein 35 (IFI35) ELISA kit) used in this application is a commercial product purchased from Shanghai Bioassay Technology Laboratory (BT-Lab) with the product number: E5498Hu.

[0263] 5. Test results and conclusions

[0264] Based on the aforementioned patient inclusion criteria, a total of 17 clinical samples were analyzed. Patient samples were divided into two groups based on pathological response (tumor cell elimination after immunotherapy): a responder group and a non-responder group. The test results are shown in Table 1.

[0265] Table 1. Test results of 17 serum samples

[0266] Note: 1. T stage (1-T1; 2-T2; 3-T3; 4-T4a; 5-T4b; 6-uncertain);

[0267] 2. N stage (0-N0; 1-N1; 2-N2a; 3-N2b; 4-N2c; 5-N3a; 6-N3b; 7-unclear; 8-N2; 9-N3);

[0268] 3. M stage (0-M0; 1-M1 (directly write the number and distant metastasis site, such as: 1 left lung>; 2-unclear);

[0269] 4. Prognostic stage grouping (1-stage I; 2-stage II; 3-stage III; 4-stage IVA; 5-stage IVB; 6-stage IVC; 7-uncertain);

[0270] 5. Pathological response of the primary lesion (0 - unresected; 1 - pCR <no residual viable cancer cells>; 2 - residual cancer cells);

[0271] 6. Pathological response of lymph nodes (0 - not cleared; 1 - pCR <all lymph nodes showed changes after treatment, with no residual viable cancer cells>; 2 - residual <lymph nodes showed changes after treatment, with some metastatic cancer>; 3 - no cancer);

[0272] 7.p16 (0-not detected; 1-positive; 2-negative);

[0273] 8. Immunotherapy drugs (0-no immunotherapy; 1-pembrolizumab / Keytruda; 2-nivolumab / Opdivo; 3-tislelizumab / tislelizumab; 4-toripalimab / Toyi; 5-sintilimab / Tyvyt; 6-camrelizumab / Erica; 7-cetuximab).

[0274] 9. Pathological response (0-non-response; 1-response).

[0275] Serum samples from HNSCC patients (HPV-negative) showed a pathological response in 5 samples and a non-response in 12 samples. The median NMI level in the 5 responding samples (from 3 patients) was 651 pg / mL (191.8, 1494), while the median NMI level in the 12 non-responding samples (from 9 patients) was almost 0 (0, 12.48). The NMI level in the responding group after immunotherapy was statistically different from that in the non-responding group (Figure 1). The results showed that after immunotherapy, if the NMI level in the serum of patients with non-HPV-related HNSCC increased, it indicated that the patient had a pathological response, that is, the immunotherapy was effective; if the NMI level remained at a low level after immunotherapy, it indicated that the patient had no pathological response, that is, the immunotherapy was ineffective.

[0276] Receiver operating characteristic (ROC) curve analysis showed that the area under the curve (AUC) for NMI in predicting pathological responses to immunotherapy in patients with non-HPV-related HNSCC reached 0.875, with a sensitivity of 100% and a specificity of 80%. This indicates that serum NMI levels are highly effective in predicting pathological responses to immunotherapy in HNSCC patients. Specifically, NMI is highly effective in evaluating the efficacy of immunotherapy for head and neck squamous cell carcinoma (HNSCC); similarly, IFP35 is also highly effective in evaluating the efficacy of immunotherapy for head and neck squamous cell carcinoma (HNSCC); and the combination of NMI and IFP35 is also highly effective in evaluating the efficacy of immunotherapy for head and neck squamous cell carcinoma (HNSCC).

[0277] 2. Application of NMI and / or IFP35 in evaluating the efficacy of different immunotherapies for different tumors

[0278] To fully validate NMI and / or IFP35 as biomarkers for evaluating the efficacy of tumor immunotherapy, the following clinical trials are further conducted:

[0279] (1) To investigate the relationship between the effects of different types of tumor immunotherapy and the levels of NMI and / or IFP35.

[0280] Experimental plan: Serum samples from patients with various tumor types were collected before, during, and after various immunotherapy treatments. The levels of NMI and IFP35 in the serum samples were detected. The relationship between the levels of NMI and IFP35 and the immunotherapy effect in patients with different tumor types before, during, and after immunotherapy was analyzed.

[0281] Among them, tumors include head and neck squamous cell carcinoma, liver cancer, colorectal cancer, bladder cancer, breast cancer, cervical cancer, prostate cancer, glioma, melanoma, pancreatic cancer, nasopharyngeal cancer, lung cancer, gastric cancer, adrenal cortical carcinoma, pararenal cortical carcinoma, anal cancer, appendix cancer, astrocytoma, atypical teratoma, rhabdoid tumor, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain tumor, bronchial tumor, Burkitt lymphoma, carcinoid tumor, heart tumor, bile duct epithelial carcinoma, chordoma, colorectal cancer, craniopharyngioma, carcinoma in situ of breast cancer, embryonal tumor, Endometrial cancer, ependymoma, esophageal cancer, olfactory neuroblastoma, intracranial germ cell tumor, extragonadal germ cell tumor, eye cancer, fallopian tube cancer, gallbladder cancer, head and neck cancer, hypopharyngeal cancer, Kaposi's sarcoma, kidney cancer, Langerhans cell histiocytosis, laryngeal cancer, lip cancer, oral cancer, Merkel cell carcinoma, malignant mesothelioma, multiple endocrine neoplasia syndrome, mycosis fungoides, sinus cancer, neuroblastoma, non-small cell lung cancer, ovarian cancer, pancreatic neuroendocrine tumor, islet cell tumor, papillomatosis, paraganglioma, nasal Sinus and nasal cavity cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pituitary tumor, pleuropulmonary blastoma, primary peritoneal cancer, retinoblastoma, salivary gland tumor, sarcoma, Sézale syndrome, skin cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, squamous cell carcinoma, testicular cancer, thymoma and thymic cancer, thyroid cancer, urethral cancer, uterine cancer, endometrial and uterine sarcoma, vaginal cancer, vascular tumor, vulvar cancer, single myeloma, B-cell acute lymphoid leukemia, T-cell acute lymphoid leukemia, acute lymphoid leukemia, chronic Myeloid leukemia, chronic lymphocytic leukemia, B-cell prolymphocytic leukemia, blastic plasmacytoid dendritic cell neoplasm, Burkitt's lymphoma, diffuse large B-cell lymphoma, follicular lymphoma, hairy cell leukemia, small cell or large cell-follicular lymphoma, MALT lymphoma, mantle cell lymphoma, marginal zone lymphoma, multiple myeloma, non-Hodgkin's lymphoma, plasmablastic lymphoma, plasmacytoid dendritic cell neoplasm, Waldenstrom's macroglobulinemia, and leukemia.

[0282] The immunotherapies covered include: monoclonal antibody therapy, immune checkpoint inhibitor therapy, adoptive cell therapy, oncolytic virus therapy, and tumor vaccine therapy.

[0283] Among them, the targets of the monoclonal antibodies are selected from CD20, HER2, VEGF / VEGFR, EGFR, CD19, FGL1, CD47, CD3, CD30, CD33, CD38, CD52, αVβ3, α5β1, FAP, Tenascin, CEA, EPCAM, PSMA, GAN-GD2, GAN-GD3, GM2, and IGF-IR.

[0284] Wherein, immune checkpoint inhibitors are inhibitors that act on T cell negative co-stimulatory (co-inhibitory) molecules and / or their respective ligands. T cell negative co-stimulatory (co-inhibitory) molecules and / or their respective ligands are selected from CTLA-4, PD-1, PD-L1, PD-L2, B7-1, B7-2, B7-H3, B7-H4, B7-H6, A2aR, IDO, TIM-3, BTLA, VISTA, TIGIT, LAG-3, CD40, CD20, CD96, CD73, CD160, STING, CEA, CD47, PVRIG, LAIR1, 2B4, KIR, CEACAM1, GARP, PS, CSF1R, CD94 / NKG2A, TDO, TNFR and DcR3.

[0285] Among them, adoptive cell therapy includes: chimeric antigen receptor T cell (CAR-T) therapy, chimeric antigen receptor NK cell (CAR-NK) therapy, T cell receptor chimeric T cell (TCR-T) therapy, tumor-infiltrating immune cell (TILs) therapy, cytokine-induced killer (CIK) cell therapy, lymphokine-activated killer (LAK) cell therapy, and natural killer (NK) cell therapy.

[0286] Among them, oncolytic viruses include alphavirus, adenovirus, vaccinia virus, Sindbis virus, Seneca Valley virus, coxsackievirus, measles virus, reovirus, cowpox virus, Newcastle disease virus, vesicular stomatitis virus, herpes simplex virus, poliovirus, influenza virus, mumps virus and parvovirus.

[0287] Among them, tumor vaccines include dendritic cell (DCs) vaccines, nucleic acid vaccines and peptide vaccines.

[0288] (2) To investigate the relationship between different immunotherapy regimens and NMI and / or IFP35 levels.

[0289] Experimental plan: Serum samples were collected from patients with various types of tumors who received different immunotherapies before, during, and after immunotherapy. The levels of NMI and / or IFP35 in the serum samples were detected, and the relationship between different immunotherapy regimens, NMI and / or IFP35 levels, and immune effects were analyzed.

[0290] 1. Clinical sample collection:

[0291] According to the above two experimental schemes, clinical samples (serum) of patients at different time points (before immunotherapy, during immunotherapy, and after immunotherapy) were collected and frozen at -80°C for future use.

[0292] Patient inclusion criteria: 1) Pathological diagnosis of cancer (see point (1) for details of tumor); 2) Immunotherapy (

[0293] For details of the immunotherapy regimen, please refer to point (1). Cancer and immunotherapy are both selected from the above-mentioned tumors and immunotherapy. For example, when verifying that NMI and / or IFP35 can be used as markers for evaluating the efficacy of tumor vaccine treatment for gastric cancer, the tumor is gastric cancer and the immunotherapy is tumor vaccine treatment; when verifying that NMI and / or IFP35 can be used as markers for evaluating the efficacy of immune checkpoint inhibitor treatment for liver cancer, the tumor is liver cancer and the immunotherapy is immune checkpoint inhibitor treatment.

[0294] 2. Detection of NMI and / or IFP35 in Serum Samples

[0295] The detection method is the same as the first point (application of NMI in evaluating the efficacy of immunotherapy for head and neck squamous cell carcinoma (HNSCC)).

[0296] 3. Results

[0297] ROC curve analysis showed that NMI and / or IFP35 can be used as markers for evaluating the efficacy of various immunotherapies for various types of tumors.

[0298] Example 2 Use of interferon-induced protein 35kD (IFP35), interferon-induced protein 35kD (IFP35) promoter, N-Myc interacting protein (NMI), and / or N-Myc interacting protein (NMI) promoter in the preparation of tumor immunotherapy enhancers

[0299] 1. Experimental purpose: This experiment intends to use an experimental animal tumor-bearing model to explore the application value of interferon-induced protein 35kD (IFP35), interferon-induced protein 35kD (IFP35) promoter, N-Myc interacting protein (NMI), and / or N-Myc interacting protein (NMI) promoter in the preparation of tumor immunotherapy enhancers.

[0300] 2. Experimental methods:

[0301] This study will be conducted for all interferon-induced protein 35kD (IFP35), interferon-induced protein 35kD (IFP35) promoters, N-Myc interacting protein (NMI), and / or N-Myc interacting protein (NMI) promoters, tumor types, and immunotherapies included below.

[0302] Among them, interferon-induced protein 35kD (IFP35) promoters include: substances that increase IFP35 levels, substances that enhance IFP35 activity, and substances that delay IFP35 metabolism.

[0303] Among them, IFP35 promoters include: natural purified substances, modified natural purified substances, semi-synthetic substances, and chemically synthesized substances.

[0304] Among them, IFP35 promoters include chemical drugs, peptide drugs, protein drugs and gene drugs.

[0305] Among them, IFP35 promoters include: biomaterials related to IFP35, exogenous IFP35, nanoparticles carrying genes (IFP35 genes), PEG-modified proteins (IFP35 proteins), protein microspheres (IFP35 proteins), liposome-encapsulated target genes (IFP35 genes) or proteins (IFP35 proteins), extracellular vesicles carrying target genes (IFP35 genes) or proteins (IFP35 proteins), and IFP35 precursor proteins or conjugates or complexes that can be converted into IFP35 in vivo.

[0306] Wherein, the biological material related to IFP35 comprises at least one of d1)-d16): d1) a nucleic acid molecule encoding IFP35; d2) an expression cassette comprising the nucleic acid molecule described in d1); d3) a vector comprising the nucleic acid molecule described in d1); d4) a vector comprising the expression cassette described in d2); d5) a transgenic cell line comprising the nucleic acid molecule described in d1); d6) a transgenic cell line comprising the expression cassette described in d2); d7) a transgenic cell line comprising the vector described in d3); d8) a transgenic cell line comprising the vector described in d4); d9) a microorganism comprising the nucleic acid molecule described in d1); d10) a microorganism comprising the expression cassette described in d2); d11) a microorganism comprising the vector described in d3); d12) a microorganism comprising the vector described in d4); d13) a virus comprising the nucleic acid molecule described in d1); d14) a virus comprising the expression cassette described in d2); d15) a virus comprising the vector described in d3); d16) a virus comprising the vector described in d4).

[0307] Among them, N-Myc interacting protein (NMI) promoters include: substances that increase NMI levels, substances that enhance NMI activity, and substances that delay NMI metabolism.

[0308] Among them, NMI promoters include: natural purified substances, modified natural purified substances, semi-synthetic substances, and chemically synthesized substances.

[0309] Among them, NMI promoters include chemical drugs, peptide drugs, protein drugs and gene drugs.

[0310] Among them, NMI promoters include: biomaterials related to NMI, exogenous NMI, nanoparticles carrying genes (NMI genes), PEG-modified proteins (NMI proteins), protein microspheres (NMI proteins), liposome-encapsulated target genes (NMI genes) or proteins (NMI proteins), extracellular vesicles carrying target genes (NMI genes) or proteins (NMI proteins), NMI precursor proteins or conjugates or complexes that can be converted into NMI in vivo.

[0311] Among them, the biological materials related to NMI include at least one of f1) to f16): f1) a nucleic acid molecule encoding NMI; f2) an expression cassette containing the nucleic acid molecule described in f1); f3) a vector containing the nucleic acid molecule described in f1); f4) a vector containing the expression cassette described in f2); f5) a transgenic cell line containing the nucleic acid molecule described in f1); f6) a transgenic cell line containing the expression cassette described in f2); f7) a transgenic cell line containing the vector described in f3); f8) a transgenic cell line containing the vector described in f4); f9) a microorganism containing the nucleic acid molecule described in f1); f10) a microorganism containing the expression cassette described in f2); f11) a microorganism containing the vector described in f3); f12) a microorganism containing the vector described in f4); f13) a virus containing the nucleic acid molecule described in f1); f14) a virus containing the expression cassette described in f2); f15) a virus containing the vector described in f3); f16) a virus containing the vector described in f4).

[0312] Among them, tumors include head and neck squamous cell carcinoma, liver cancer, colorectal cancer, bladder cancer, breast cancer, cervical cancer, prostate cancer, glioma, melanoma, pancreatic cancer, nasopharyngeal cancer, lung cancer, gastric cancer, adrenal cortical carcinoma, pararenal cortical carcinoma, anal cancer, appendix cancer, astrocytoma, atypical teratoma, rhabdoid tumor, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain tumor, bronchial tumor, Burkitt lymphoma, carcinoid tumor, heart tumor, bile duct epithelial carcinoma, chordoma, colorectal cancer, cranial cancer, Pharyngioma, ductal carcinoma in situ, embryonal tumor, endometrial cancer, ependymoma, esophageal cancer, olfactory neuroblastoma, intracranial germ cell tumor, extragonadal germ cell tumor, eye cancer, fallopian tube cancer, gallbladder cancer, head and neck cancer, hypopharyngeal cancer, Kaposi's sarcoma, kidney cancer, Langerhans cell histiocytosis, laryngeal cancer, lip cancer, oral cancer, Merkel cell carcinoma, malignant mesothelioma, multiple endocrine neoplasia syndrome, mycosis fungoides, nasal sinus cancer, neuroblastoma, non-small cell lung cancer, ovarian cancer Ovarian cancer, pancreatic neuroendocrine tumors, islet cell tumors, papillomatosis, paraganglioma, sinonasal cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pituitary tumors, pleuropulmonary blastoma, primary peritoneal cancer, retinoblastoma, salivary gland tumors, sarcomas, Sézale syndrome, skin cancer, small cell lung cancer, small bowel cancer, soft tissue sarcoma, squamous cell carcinoma, testicular cancer, thymoma and thymic cancer, thyroid cancer, urethral cancer, uterine cancer, endometrial and uterine sarcoma, vaginal cancer, vascular Tumors, vulvar cancer, single myeloma, B-cell acute lymphoid leukemia, T-cell acute lymphoid leukemia, acute lymphoid leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia, B-cell prolymphocytic leukemia, blastic plasmacytoid dendritic cell neoplasm, Burkitt's lymphoma, diffuse large B-cell lymphoma, follicular lymphoma, hairy cell leukemia, small cell or large cell-follicular lymphoma, MALT lymphoma, mantle cell lymphoma, marginal zone lymphoma, multiple myeloma, non-Hodgkin's lymphoma, plasmablastic lymphoma, plasmacytoid dendritic cell neoplasm, Waldenstrom's macroglobulinemia, and leukemia.

[0313] The immunotherapies covered include: monoclonal antibody therapy, immune checkpoint inhibitor therapy, adoptive cell therapy, oncolytic virus therapy, and tumor vaccine therapy.

[0314] Among them, the targets of the monoclonal antibodies are selected from CD20, HER2, VEGF / VEGFR, EGFR, CD19, FGL1, CD47, CD3, CD30, CD33, CD38, CD52, αVβ3, α5β1, FAP, Tenascin, CEA, EPCAM, PSMA, GAN-GD2, GAN-GD3, GM2, and IGF-IR.

[0315] Wherein, immune checkpoint inhibitors are inhibitors that act on T cell negative co-stimulatory (co-inhibitory) molecules and / or their respective ligands. T cell negative co-stimulatory (co-inhibitory) molecules and / or their respective ligands are selected from CTLA-4, PD-1, PD-L1, PD-L2, B7-1, B7-2, B7-H3, B7-H4, B7-H6, A2aR, IDO, TIM-3, BTLA, VISTA, TIGIT, LAG-3, CD40, CD20, CD96, CD73, CD160, STING, CEA, CD47, PVRIG, LAIR1, 2B4, KIR, CEACAM1, GARP, PS, CSF1R, CD94 / NKG2A, TDO, TNFR and DcR3.

[0316] Among them, adoptive cell therapy includes: chimeric antigen receptor T cell (CAR-T) therapy, chimeric antigen receptor NK cell (CAR-NK) therapy, T cell receptor chimeric T cell (TCR-T) therapy, tumor-infiltrating immune cell (TILs) therapy, cytokine-induced killer (CIK) cell therapy, lymphokine-activated killer (LAK) cell therapy, and natural killer (NK) cell therapy.

[0317] Among them, oncolytic viruses include alphavirus, adenovirus, vaccinia virus, Sindbis virus, Seneca Valley virus, coxsackievirus, measles virus, reovirus, cowpox virus, Newcastle disease virus, vesicular stomatitis virus, herpes simplex virus, poliovirus, influenza virus, mumps virus and parvovirus.

[0318] Among them, tumor vaccines include dendritic cell (DCs) vaccines, nucleic acid vaccines and peptide vaccines.

[0319] Experiments were performed on all interferon-induced protein 35kD (IFP35), interferon-induced protein 35kD (IFP35) promoters, N-Myc interacting protein (NMI), and / or N-Myc interacting protein (NMI) promoters, tumor types and immunotherapies included above.

[0320] 1. Case I for NMI or IFP35 as a monoclonal antibody therapy enhancer is as follows:

[0321] 1. Experimental groups: This experiment will set up the following six groups: 1) wild-type mouse tumor-bearing placebo treatment group, 2) wild-type mouse tumor-bearing monoclonal antibody treatment group, 3) wild-type mouse tumor-bearing monoclonal antibody combined with NMI recombinant protein treatment group, 4) wild-type mouse tumor-bearing monoclonal antibody combined with IFP35 recombinant protein treatment group, 5) NMI knockout mouse tumor-bearing monoclonal antibody treatment group, 6) IFP35 knockout mouse tumor-bearing monoclonal antibody treatment group.

[0322] 2. Drug Selection and Tumor Model Construction: To investigate the enhancement of NMI / IFP35 in response to immune checkpoint inhibitors, this study used amivantamab, an FDA-approved antibody targeting EGFR / MET for the treatment of non-small cell lung cancer, to treat a Lewis lung cancer tumor model in mice. Lewis cells were cultured in vitro and passaged to an appropriate cell suspension. Under sterile conditions, the cell concentration was adjusted to 3×10 cells / mL using PBS. 6 / mL, Lewis tumor cell suspension was sterilely inoculated subcutaneously on the back of 6-8 week old female BALB / c mice with an inoculation volume of 100 μL and a tumor cell inoculation volume of 3×10 5 / Only.

[0323] 3. Tumor growth assessment: Starting from the 6th day of tumor bearing, the tumor volume was measured every other day. The longest and shortest diameters of the tumor were measured with a vernier caliper. Tumor volume = tumor long diameter × tumor short diameter 2 / 2; when the tumor volume reaches 100mm 3 When the tumor is successfully loaded.

[0324] 4. Treatment plan: When the average tumor volume reaches about 60mm 3 At the time of treatment, tumor-bearing mice were evenly divided into groups to ensure that the average tumor volume of each group of mice remained consistent before treatment. On days 0, 3, 7, 10, 14, and 17 after grouping, mice were given amivantamab antibodies by intraperitoneal injection at a dose of 10 mg / kg of mouse body weight. The placebo treatment group was intraperitoneally injected with an equal volume of PBS. The NMI / IFP35 combination treatment group was given simultaneous administration of the amivantamab antibody and the biologically active NMI / IFP35 recombinant protein was administered intratumorally at a dose of 100 μg / mouse. Tumor volume and mouse body weight were measured every other day after the start of treatment to evaluate the therapeutic efficacy. The treatment cycle was completed on the second day after the injection of the amivantamab antibody. The tumor-bearing mice were humanely sacrificed, and the tumor tissue was completely collected, weighed, and recorded.

[0325] Case II for NMI or IFP35 as a potentiator of immune checkpoint inhibitor therapy is as follows:

[0326] 1. Experimental groups: This experiment will set up the following six groups: 1) wild-type mice bearing tumors and placebo treatment group, 2) wild-type mice bearing tumors and immune checkpoint inhibitor treatment group, 3) wild-type mice bearing tumors and immune checkpoint inhibitor combined with NMI recombinant protein treatment group, 4) wild-type mice bearing tumors and immune checkpoint inhibitor combined with IFP35 recombinant protein treatment group, 5) NMI knockout mice bearing tumors and immune checkpoint inhibitor treatment group, 6) IFP35 knockout mice bearing tumors and immune checkpoint inhibitor treatment group.

[0327] 2. Drug Selection and Tumor Model Construction: To investigate the enhancement of NMI / IFP35 in response to immune checkpoint inhibitors, this study used the most popular PD-1 antibody in clinical practice to treat the CT26 mouse colon cancer tumor model, which has a known high response rate to PD-1 neutralizing antibodies. CT26 cells were cultured in vitro and passaged to an appropriate state before a cell suspension was prepared. Under sterile conditions, the cell concentration was adjusted to 3×10 cells using PBS. 6 / mL, CT26 tumor cell suspension was sterilely inoculated subcutaneously on the back of 6-8 week old female BALB / c mice, with an inoculation volume of 100 μL and a tumor cell inoculation volume of 3×10 5 / Only.

[0328] 3. Tumor growth assessment: Starting from the 6th day of tumor bearing, the tumor volume was measured every other day. The longest and shortest diameters of the tumor were measured with a vernier caliper. Tumor volume = tumor long diameter × tumor short diameter 2 / 2; when the tumor volume reaches 100mm 3 When the tumor is successfully loaded.

[0329] 4. Treatment plan: When the average tumor volume reaches about 60mm 3 At the time of treatment, tumor-bearing mice were divided into groups to ensure that the average tumor volume of each group of mice remained consistent before treatment. On days 0, 3, 7, 10, 14, and 17 after grouping, mice were given anti-PD-1 neutralizing antibodies by intraperitoneal injection at a dose of 10 mg / kg of mouse body weight. The placebo treatment group was intraperitoneally injected with an equal volume of PBS. The NMI / IFP35 combination treatment group was given simultaneous administration of the PD-1 neutralizing antibody and the biologically active NMI / IFP35 recombinant protein at a dose of 100 μg / mouse by intratumoral administration. Tumor volume and mouse body weight were measured every other day after the start of treatment to evaluate the therapeutic efficacy. The treatment cycle was completed on the second day after the injection of the PD-1 inhibitor. The tumor-bearing mice were humanely sacrificed, and the tumor tissue was completely collected, weighed, and recorded.

[0330] Case III: NMI or IFP35 as a CAR-T cell adoptive therapy enhancer

[0331] 1. Experimental groups: This experiment will set up the following six groups: 1) wild-type mouse tumor-bearing placebo treatment group, 2) wild-type mouse tumor-bearing CAR-T cell adoptive therapy group, 3) wild-type mouse tumor-bearing CAR-T cell adoptive therapy combined with NMI recombinant protein treatment group, 4) wild-type mouse tumor-bearing CAR-T cell adoptive therapy combined with IFP35 recombinant protein treatment group, 5) NMI knockout mouse tumor-bearing CAR-T cell adoptive therapy group, and 6) IFP35 knockout mouse tumor-bearing CAR-T cell adoptive therapy group.

[0332] 2. Drug Selection and Tumor Model Construction: To investigate the enhancement of NMI / IFP35 in adoptive CAR-T cell therapy, this study used a classic CD19-targeted CAR-T cell adoptive therapy in a mouse acute B-lymphoblastic leukemia Nalm-6 tumor-bearing model. Nalm-6 cells were cultured in vitro and passaged to an appropriate state before a cell suspension was prepared. Under sterile conditions, the cell concentration was adjusted to 3×10 cells / mL using PBS. 6 / mL, Nalm-6 tumor cell suspension was intravenously infused into 6-8 week old female NOG mice, with an inoculation volume of 100 μL and a tumor cell inoculation volume of 3×10 5 / Only.

[0333] 3.CAR-T cell construction: peripheral blood of mice was collected and PBMC cells were isolated. CD3 + T cells. After transfecting the target gene into T cells using CEA-CAR lentiviral transfection, qRT-PCR was used to detect target gene expression and verify transfection efficiency.

[0334] 4. Leukemia Progress Assessment: Because leukemia is a non-solid tumor, the detection method primarily focuses on the number of cancer cells in the mouse peripheral blood. Orbital blood was collected on day 6 of tumor loading, and the number of Nalm-6 cancer cells in the peripheral blood was determined by flow cytometry. After adoptive transfer of CAR-T cells, peripheral blood was collected every 5 days for flow cytometry.

[0335] 5. Treatment plan: 7 days after the infusion of Nalm-6 cells, the tumor-bearing mice were divided into groups according to the number of Nalm-6 cells in the peripheral blood, ensuring that the average number of Nalm-6 cells in each group was consistent before treatment. After grouping, 1×10 7 The placebo group was injected with an equal volume of PBS. The NMI / IFP35 combination treatment group received 100 μg of the biologically active NMI / IFP35 recombinant protein intratumorally, simultaneously with the CAR-T cells. Mice were weighed every other day after the start of treatment, and peripheral blood was collected every five days for testing to assess therapeutic efficacy. The treatment cycle was completed on day 17 after adoptive transfer of CAR-T cells. Tumor-bearing mice were humanely sacrificed, and tumor tissue was fully harvested, weighed, and recorded.

[0336] Case IV for NMI or IFP35 as an enhancer of oncolytic virus therapy is as follows:

[0337] 1. Experimental groups: This experiment will set up the following six groups: 1) wild-type mouse tumor-bearing placebo treatment group, 2) wild-type mouse tumor-bearing oncolytic virus treatment group, 3) wild-type mouse tumor-bearing oncolytic virus combined with NMI recombinant protein treatment group, 4) wild-type mouse tumor-bearing oncolytic virus combined with IFP35 recombinant protein treatment group, 5) NMI knockout mouse tumor-bearing oncolytic virus treatment group, 6) IFP35 knockout mouse tumor-bearing oncolytic virus treatment group.

[0338] 2. Drug Selection and Tumor Model Construction: To investigate the enhancement of NMI / IFP35 in oncolytic virus therapy, this study used T-VEC, an FDA-approved and clinically used oncolytic virus for the treatment of melanoma, to treat a mouse skin melanoma B16 tumor model. B16 cells were cultured in vitro and passaged to an appropriate state before a cell suspension was prepared. Under sterile conditions, the cell concentration was adjusted to 3×10 cells / mL using PBS. 6 / mL, B16 tumor cell suspension was sterilely inoculated subcutaneously on the back of 6-8 week old female C57BL / 6J mice, with an inoculation volume of 100 μL and a tumor cell inoculation volume of 3×10 5 / Only.

[0339] 3. Tumor growth assessment: Starting from the 6th day of tumor bearing, the tumor volume was measured every other day. The longest and shortest diameters of the tumor were measured with a vernier caliper. Tumor volume = tumor long diameter × tumor short diameter 2 / 2; when the tumor volume reaches 100mm 3 When the tumor is successfully loaded.

[0340] 4. Treatment plan: When the average tumor volume reaches about 60mm 3 The tumor-bearing mice were divided into groups to ensure that the average tumor volume of each group of mice before treatment was consistent. The administration method was intratumoral injection of 1×10 7 PFU / 100μL oncolytic virus. Before administration, the virus was filtered and sterilized through a 0.22μm filter membrane and administered once a day for three consecutive days. After one day off, the drug was administered for another three consecutive days. The placebo treatment group was injected with an equal volume of PBS. The NMI / IFP35 combination treatment group was administered synchronously with the oncolytic virus, and the biologically active NMI / IFP35 recombinant protein was administered intratumorally at a dose of 100μg / mouse. The tumor volume and mouse body weight were measured every other day after the start of treatment to evaluate the therapeutic efficacy. The treatment cycle was completed on the second day after the injection of the oncolytic virus. The tumor-bearing mice were humanely killed, and the tumor tissue was completely collected, weighed and recorded.

[0341] 5. Case V for NMI or IFP35 as a tumor vaccine therapy enhancer is as follows:

[0342] 1. Experimental groups: This experiment will set up the following six groups: 1) wild-type mouse tumor-bearing placebo treatment group, 2) wild-type mouse tumor vaccine treatment group, 3) wild-type mouse tumor vaccine combined with NMI recombinant protein treatment group, 4) wild-type mouse tumor-bearing tumor vaccine combined with IFP35 recombinant protein treatment group, 5) NMI knockout mouse tumor-bearing tumor vaccine treatment group, 6) IFP35 knockout mouse tumor-bearing tumor vaccine treatment group.

[0343] 2. Drug Selection and Tumor Model Construction: To investigate the enhancement of NMI / IFP35 in tumor vaccine therapy, this study used Provenge, a PDA-approved and clinically used tumor vaccine for resistant prostate cancer, to treat the RM-1 prostate cancer mouse tumor model. RM-1 cells were cultured in vitro and passaged to an appropriate cell suspension. Under sterile conditions, the cell concentration was adjusted to 3×10 cells in PBS. 6 / mL, RM-1 tumor cell suspension was sterilely inoculated subcutaneously on the back of 6-8 week old male Balb / c mice, with an inoculation volume of 100 μL and a tumor cell inoculation volume of 3×10 5 / Only.

[0344] 3. Tumor growth assessment: Starting from the 6th day of tumor bearing, the tumor volume was measured every other day. The longest and shortest diameters of the tumor were measured with a vernier caliper. Tumor volume = tumor long diameter × tumor short diameter 2 / 2; when the tumor volume reaches 100mm 3 When the tumor is successfully loaded.

[0345] 4. Treatment plan: When the average tumor volume reaches about 60mm 3 When the tumor-bearing mice were treated, they were divided into groups evenly to ensure that the average tumor volume of each group of mice remained consistent before treatment. The vaccine was administered subcutaneously in the right flank of the mice for a total of three injections, with an interval of two days between each injection. The placebo treatment group was injected with an equal volume of PBS. The NMI / IFP35 combination treatment group was administered synchronously with the vaccine, and the biologically active NMI / IFP35 recombinant protein was administered intratumorally at a dose of 100 μg / mouse. The tumor volume and mouse body weight were measured every other day after the start of treatment to evaluate the therapeutic efficacy. The treatment cycle was completed on the 24th day after the completion of the vaccination. The tumor-bearing mice were humanely killed, and the tumor tissue was completely collected, weighed and recorded.

[0346] The experimental grouping and evaluation criteria implemented in the above cases apply to all tumor types and immunotherapies covered by this patent. The specific implementation plan will be adjusted based on the growth characteristics of different tumors and the response rates of different therapies. The principle is to select the most clinically applicable and optimal immunotherapy for that tumor type, and for each immunotherapy, select the tumor type with the highest response rate.

[0347] The results of the above case are as follows:

[0348] Compared with the use of immunotherapy alone in the wild-type mouse tumor-bearing model, the tumor treatment efficacy of the mouse group treated with the combined NMI / IFP35 biological agent was better. It can be judged that the NMI / IFP35 active biological agent has application value in the development of tumor immunotherapy enhancers.

[0349] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be considered as equivalent replacement methods and are included in the scope of protection of the present invention.

Claims

1. Use of a substance for detecting a marker in preparing a product, wherein the marker comprises the IFP35 protein family, wherein the IFP35 protein family comprises interferon-induced protein 35kD (IFP35) and / or N-Myc interacting protein (NMI); the product is used for at least one of (1)-(3): (1) evaluation of the efficacy of tumor immunotherapy; (2) prognosis evaluation of tumor immunotherapy; (3) selection of treatment regimen.

2. The use according to claim 1, characterized in that: The substance for detecting the marker comprises a substance for detecting the marker at the gene level and / or protein level; preferably, the substance comprises a substance for one or more detection techniques or methods selected from the group consisting of: immunohistochemistry, Western blotting, Northern blotting, PCR, biochip, nucleic acid sequencing, amino acid sequencing; preferably, the immunohistochemistry is selected from at least one of immunofluorescence, immunoenzyme-linked immunosorbent assay (ELISA) and immunocolloidal gold assay; preferably, the substance for detecting the marker is selected from one or more of a substance specific to the marker, a probe specific to the marker, a gene chip, and a PCR primer; preferably, the substance specific to the marker comprises any one of (a1) to (a3): (a1) an antibody that specifically binds to the marker; (a2) a ligand protein or polypeptide that specifically binds to the marker; (a3) ​​a non-protein compound that specifically recognizes the marker; preferably, the antibody comprises at least one of a polyclonal antibody, a monoclonal antibody, a single-chain antibody, a functional antibody fragment, an antibody Fab region, a nanobody, a chimeric antibody, and a multispecific antibody.

3. The use according to claim 1, characterized in that: The product comprises at least one of a reagent, a kit, a test paper, a chip, and a system; preferably, the test sample of the product is selected from at least one of the body fluids, tissues, cells, and excretions of the subject to be tested; preferably, the body fluid comprises at least one of blood and lymph; preferably, the blood comprises at least one of serum, plasma, dried blood spots, and whole blood; preferably, the tissue comprises tumor tissue; preferably, the excretion comprises at least one of urine, feces, and tears; preferably, the subject to be tested comprises mammals; preferably, the subject to be tested comprises humans.

4. The use according to any one of claims 1 to 3, characterized in that: The markers also include other markers used for evaluating the efficacy of tumor immunotherapy, evaluating the prognosis of tumor immunotherapy, and / or selecting a treatment regimen.

5. Use of (c1) and / or (c2) in the preparation of tumor immunotherapy enhancers; (c1) interferon-induced protein 35kD (IFP35) and / or interferon-induced protein 35kD (IFP35) promoter; (c2) N-Myc interacting protein (NMI) and / or N-Myc interacting protein (NMI) promoter.

6. The use according to claim 5, characterized in that: The interferon-induced protein 35kD (IFP35) promoter comprises: at least one of a substance that increases the level of IFP35, a substance that enhances the activity of IFP35, and a substance that delays the metabolism of IFP35; and / or the N-Myc interacting protein (NMI) promoter comprises: at least one of a substance that increases the level of NMI, a substance that enhances the activity of NMI, and a substance that delays the metabolism of NMI.

7. The use according to claim 6, characterized in that: The IFP35 promoter comprises: at least one of natural purified substances, modified natural purified substances, semi-synthetic substances, and chemically synthesized substances; preferably, the IFP35 promoter comprises at least one of chemical drugs, polypeptide drugs, protein drugs, and gene drugs; preferably, the IFP35 promoter comprises: at least one of biological materials related to IFP35, exogenous IFP35, nanoparticles carrying genes, PEG-modified proteins, protein microspheres, liposomes encapsulating target genes or proteins, extracellular vesicles carrying target genes or proteins, and IFP35 precursor proteins or conjugates or complexes that can be converted into IFP35 in vivo; Preferably, the biological material related to IFP35 comprises at least one of d1)-d16): d1) a nucleic acid molecule encoding IFP35; d2) an expression cassette comprising the nucleic acid molecule of d1); d3) a vector comprising the nucleic acid molecule of d1); d4) a vector comprising the nucleic acid molecule of d2). d5) a transgenic cell line comprising the nucleic acid molecule described in d1); d6) a transgenic cell line comprising the expression cassette described in d2); d7) a transgenic cell line comprising the vector described in d3); d8) a transgenic cell line comprising the vector described in d4); d9) a microorganism comprising the nucleic acid molecule described in d1); d10) a microorganism comprising the expression cassette described in d2); d11) a microorganism comprising the vector described in d3); d12) a microorganism comprising the vector described in d4); d13) a virus comprising the nucleic acid molecule described in d1); d14) a virus comprising the expression cassette described in d2); d15) a virus comprising the vector described in d3); d16) a virus comprising the vector described in d4); Preferably, the IFP35 is human IFP35; Preferably, the NMI promoter comprises: at least one of natural purified substances, modified natural purified substances, semi-synthetic substances, and chemically synthesized substances; preferably, the NMI promoter comprises at least one of chemical drugs, polypeptide drugs, protein drugs, and gene drugs; preferably, the NMI promoter comprises: at least one of biological materials related to NMI, exogenous NMI, nanoparticles carrying genes, PEG-modified proteins, protein microspheres, liposomes encapsulating target genes or proteins, extracellular vesicles carrying target genes or proteins, and NMI precursor proteins or conjugates or complexes that can be converted into NMI in vivo; Preferably, the biological material associated with NMI comprises at least one of f1)-f16): f1) a nucleic acid molecule encoding NMI; f2) an expression cassette comprising the nucleic acid molecule of f1); f3) a vector comprising the nucleic acid molecule of f1); f4) a vector comprising the expression cassette of f2); f5) a transgenic cell line comprising the nucleic acid molecule of f1); f6) a transgenic cell line comprising the expression cassette of f2); f7) a transgenic cell line comprising the vector of f3); f8) a transgenic cell line comprising the vector of f4); f9) a microorganism comprising the nucleic acid molecule of f1); f10) a microorganism comprising the expression cassette of f2); f11) a microorganism comprising the vector of f3); f12) a microorganism comprising the vector of f4); f13) a virus comprising the nucleic acid molecule of f1); f14) a virus comprising the expression cassette of f2); f15) a virus comprising the vector of f3); f16) a virus comprising the vector of f4); Preferably, the NMI is a human NMI.

8. The use according to any one of claims 5 to 7, characterized in that: The tumor immunotherapy enhancer also contains a pharmaceutically acceptable carrier or excipient; preferably, the form of the tumor immunotherapy enhancer makes it suitable for administration in a manner selected from the following groups: oral administration, injection (such as direct naked DNA or protein injection, liposome-encapsulated DNA, RNA or protein injection), gold-coated gene gun bombardment, reproduction-defective bacteria carrying plasmid DNA, replication-defective adenovirus carrying target DNA or target gene encoded protein, electroporation, intravenous, pulmonary, mucosal, nasal, intraperitoneal, intracranial, intratumoral, sublingual, buccal, transdermal administration; preferably, the dosage form of the tumor immunotherapy enhancer contains at least one of capsules, tablets, microcapsule preparations, injections, suppositories, sprays, powders, soft capsules, dripping pills, honey pills, pills, granules, honey-refined pastes, sustained-release preparations, oral liquid preparations, chewable tablets, buccal tablets, transdermal patches, and effervescent tablets; preferably, the tumor immunotherapy enhancer also contains other substances that improve the efficacy of tumor immunotherapy.

9. A tumor immunotherapy enhancer, comprising: (c1) at least one of interferon-induced protein 35kD (IFP35) and interferon-induced protein 35kD (IFP35) promoter; and / or (c2) at least one of N-Myc interacting protein (NMI) and N-Myc interacting protein (NMI promoter).

10. The tumor immunotherapy enhancer according to claim 9, characterized in that: The tumor immunotherapy enhancer comprises (c1) and (c2); preferably, the interferon-induced protein 35kD (IFP35) promoter is the interferon-induced protein 35kD (IFP35) promoter described in any one of claims 6-8; preferably, the N-Myc interacting protein (NMI) promoter is the N-Myc interacting protein (NMI) promoter described in any one of claims 6-8.

11. The tumor immunotherapy enhancer according to claim 9 or 10, characterized in that: The tumor immunotherapy enhancer also contains a pharmaceutically acceptable carrier or excipient; preferably, the form of the tumor immunotherapy enhancer makes it suitable for administration in a manner selected from the following groups: oral administration, injection (such as direct naked DNA or protein injection, liposome-encapsulated DNA, RNA or protein injection), gold-coated gene gun bombardment, reproduction-defective bacteria carrying plasmid DNA, replication-defective adenovirus carrying target DNA or target gene encoded protein, electroporation, intravenous, pulmonary, mucosal, nasal, intraperitoneal, intracranial, intratumoral, sublingual, buccal, transdermal administration; preferably, the dosage form of the tumor immunotherapy enhancer contains at least one of capsules, tablets, microcapsule preparations, injections, suppositories, sprays, powders, soft capsules, dripping pills, honey pills, pills, granules, honey-refined pastes, sustained-release preparations, oral liquid preparations, chewable tablets, buccal tablets, transdermal patches, and effervescent tablets; preferably, the tumor immunotherapy enhancer also contains other substances that improve the efficacy of tumor immunotherapy.

12. An anti-tumor pharmaceutical composition comprising: (e1) a tumor immunotherapy enhancer; and (e2) a tumor immunotherapy agent; the tumor immunotherapy enhancer is the tumor immunotherapy enhancer according to any one of claims 9 to 11.

13. The anti-tumor pharmaceutical composition according to claim 12, characterized in that: The pharmaceutical composition further comprises a pharmaceutically acceptable carrier or excipient; preferably, the form of the pharmaceutical composition makes it suitable for administration in a manner selected from the group consisting of: oral administration, injection (e.g., direct naked DNA or protein injection, liposome-encapsulated DNA, RNA or protein injection), gold-coated gene gun bombardment, reproduction-defective bacteria carrying plasmid DNA, replication-defective adenovirus carrying target DNA or target gene-encoded protein, electroporation, intravenous, pulmonary, mucosal, nasal, intraperitoneal, intracranial, intratumoral, sublingual, buccal, transdermal administration; preferably, the dosage form of the pharmaceutical composition comprises at least one of capsules, tablets, microcapsule preparations, injections, suppositories, sprays, powders, soft capsules, dripping pills, honey pills, pills, granules, honey-refined pastes, sustained-release preparations, oral liquid preparations, chewable tablets, buccal tablets, transdermal patches, and effervescent tablets; preferably, the pharmaceutical composition further comprises instructions; preferably, the instructions record the number of administrations, dosage, and administration method.

14. An anti-tumor drug kit, comprising: (e1) a tumor immunotherapy enhancer; and (e2) a tumor immunotherapy agent; the tumor immunotherapy enhancer is the tumor immunotherapy enhancer according to any one of claims 9 to 11.

15. The anti-tumor drug set according to claim 14, characterized in that: The tumor immunotherapy enhancer and the tumor immunotherapy agent exist independently; preferably, the tumor immunotherapy agent further comprises a pharmaceutically acceptable carrier or excipient; preferably, the tumor immunotherapy agent is in a form suitable for administration selected from the following groups: oral administration, injection (e.g., direct naked DNA or protein injection, liposome-encapsulated DNA, RNA or protein injection), gold-coated gene gun bombardment, replication-defective bacteria carrying plasmid DNA, replication-defective adenovirus carrying target DNA or target gene-encoded protein, electroporation, intravenous administration, pulmonary administration, mucosal administration, nasal cavity, intraperitoneal administration, intracranial administration , intratumoral, sublingual, buccal, and transdermal administration; preferably, the dosage form of the tumor immunotherapy agent includes at least one of capsules, tablets, microcapsule preparations, injections, suppositories, sprays, powders, soft capsules, dripping pills, honey pills, pills, granules, honey-refined pastes, sustained-release preparations, oral liquid preparations, chewable tablets, buccal tablets, transdermal patches, and effervescent tablets; preferably, the tumor immunotherapy enhancer and the tumor immunotherapy agent are administered simultaneously or in any order; preferably, the drug set also includes instructions; preferably, the instructions record the number of administrations, dosage, and administration method.

16. Use of a tumor immunotherapy enhancer and a tumor immunotherapy agent in the preparation of a drug for treating tumors, wherein the tumor immunotherapy enhancer is the tumor immunotherapy enhancer according to any one of claims 9 to 11.

17. The anti-tumor pharmaceutical composition, anti-tumor pharmaceutical set, and use according to any one of claims 12 to 16, characterized in that: the tumor immunotherapy agent comprises: at least one of a monoclonal antibody, an immune checkpoint inhibitor, an immune cell, an oncolytic virus, and a tumor vaccine; Preferably, the target of the monoclonal antibody is selected from one of CD20, HER2, VEGF / VEGFR, EGFR, CD19, FGL1, CD47, CD3, CD30, CD33, CD38, CD52, αVβ3, α5β1, FAP, Tenascin, CEA, EPCAM, PSMA, GAN-GD2, GAN-GD3, GM2, and IGF-IR; Preferably, the immune checkpoint inhibitor is an inhibitor that acts on T cell negative co-stimulatory (co-inhibitory) molecules and / or their respective ligands; Preferably, the negative co-stimulatory (co-inhibitory) molecules acting on T cells and / or their respective ligands are selected from one or more of CTLA-4, PD-1, PD-L1, PD-L2, B7-1, B7-2, B7-H3, B7-H4, B7-H6, A2aR, IDO, TIM-3, BTLA, VISTA, TIGIT, LAG-3, CD40, CD20, CD96, CD73, CD160, STING, CEA, CD47, PVRIG, LAIR1, 2B4, KIR, CEACAM1, GARP, PS, CSF1R, CD94 / NKG2A, TDO, TNFR and DcR3; Preferably, the inhibitor acting on T cell negative co-stimulatory (co-inhibitory) molecules and / or their respective ligands comprises any one of (b1) to (b3): ​​(b1) antibodies that specifically bind (neutralize) T cell negative co-stimulatory (co-inhibitory) molecules and / or their respective ligands; (b2) ligand proteins or polypeptides that specifically bind (neutralize) T cell negative co-stimulatory (co-inhibitory) molecules and / or their respective ligands; (b3) non-protein compounds that specifically bind (neutralize) T cell negative co-stimulatory (co-inhibitory) molecules and / or their respective ligands; Preferably, the immune cells include: chimeric antigen receptor T cells (CAR-T), chimeric antigen receptor NK cells (CAR-NK), T cell receptor chimeric T cells (TCR-T), tumor infiltrating immune cells (TILs), cytokine-induced killer (CIK) cells, lymphokine-activated killer (LAK) cells, and natural killer (NK) cells. Preferably, the oncolytic virus comprises at least one of alphavirus, adenovirus, vaccinia virus, Sindbis virus, Seneca Valley virus, coxsackie virus, measles virus, reovirus, vaccinia virus, Newcastle disease virus, vesicular stomatitis virus, herpes simplex virus, polio virus, influenza virus, mumps virus and parvovirus; further comprises at least one of alphavirus, adenovirus, vaccinia virus, measles virus, vesicular stomatitis virus, and herpes simplex virus; Preferably, the alphavirus comprises at least one of an M1 virus and a Geta virus; Preferably, the tumor vaccine comprises at least one of a dendritic cell (DCs) vaccine, a nucleic acid vaccine and a polypeptide vaccine.

18. A marker combination comprising the IFP35 protein family, wherein the IFP35 protein family comprises interferon-induced protein 35kD (IFP35) and N-Myc interacting protein (NMI).

19. The marker combination according to claim 18, characterized in that: The marker combination is used for at least one of (1)-(3): (1) evaluation of the efficacy of tumor immunotherapy; (2) evaluation of the prognosis of tumor immunotherapy; (3) selection of treatment regimen; Preferably, the marker combination is from at least one of the body fluids, tissues, cells, and excretions of the subject to be tested; preferably, the body fluid comprises at least one of blood and lymph; preferably, the blood comprises at least one of serum, plasma, dried blood spots, and whole blood; preferably, the tissue comprises tumor tissue; preferably, the excretion comprises at least one of urine, feces, and tears; preferably, the subject to be tested comprises mammals; preferably, the subject to be tested comprises humans; preferably, the markers also comprise other markers for evaluating the efficacy of tumor immunotherapy, evaluating the prognosis of tumor immunotherapy, and / or selecting a treatment regimen.

20. A product comprising a substance for detecting the marker combination according to claim 18 or 19.

21. The product according to claim 20, characterized in that: The product is used for at least one of (1)-(3): (1) evaluation of the efficacy of tumor immunotherapy; (2) prognosis evaluation of tumor immunotherapy; (3) selection of treatment regimen; Preferably, the substance for detecting the marker combination comprises a substance for detecting the marker combination at the gene level and / or protein level; preferably, the substance comprises a substance for one or more detection techniques or methods selected from the group consisting of: immunohistochemistry, Western blotting, Northern blotting, PCR, biochip, nucleic acid sequencing, amino acid sequencing; preferably, the immunohistochemistry is selected from at least one of: immunofluorescence, ELISA and immunocolloidal gold; preferably, the substance for detecting the marker is selected from: a substance specific to the marker combination, a substance specific to the marker combination, one or more of a probe, a gene chip, and a PCR primer; preferably, the substance specific to the marker combination comprises any one of (a1) to (a3): (a1) an antibody that specifically binds to the marker combination; (a2) a ligand protein or polypeptide that specifically binds to the marker combination; (a3) ​​a non-protein compound that specifically recognizes the marker combination; preferably, the antibody comprises at least one of a polyclonal antibody, a monoclonal antibody, a single-chain antibody, a functional antibody fragment, an antibody Fab region, a nanobody, a chimeric antibody, and a multispecific antibody; preferably, the product comprises at least one of a reagent, a kit, a test strip, a chip, and a system.

22. A method for constructing a model with poor efficacy of tumor immunotherapy, which reduces the content and / or activity of the IFP35 protein family in the model, wherein the IFP35 protein family includes interferon-induced protein 35kD (IFP35) and / or N-Myc interacting protein (NMI).

23. The construction method according to claim 22, characterized in that: The method for reducing the content and / or activity of the IFP35 protein family in the model is to knock out the gene of the IFP35 protein family.

24. A method for constructing a model with good tumor immunotherapy efficacy, which increases the content and / or activity of the IFP35 protein family in the model, wherein the IFP35 protein family includes interferon-induced protein 35kD (IFP35) and / or N-Myc interacting protein (NMI).

25. The construction method according to claim 24, characterized in that: The method for increasing the content and / or activity of the IFP35 protein family in the model is to administer the tumor immunotherapy enhancer according to any one of claims 9 to 11 to the model.

26. The construction method according to any one of claims 22 to 25, characterized in that: The model comprises an animal model; preferably, the animal comprises at least one of rats and mice.

27. A system, comprising: a detection device for the content and / or expression of a marker in a test sample: used to detect the content and / or expression of a marker in a test sample; an evaluation or selection device: evaluating the efficacy of tumor immunotherapy, evaluating the prognosis of tumor immunotherapy, and / or selecting a treatment plan according to the content and / or expression of the marker; a result output device: outputting the evaluation results of the efficacy of tumor immunotherapy, the prognosis of tumor immunotherapy, and / or the selection results of a treatment plan; The markers include the IFP35 protein family, and the IFP35 protein family includes interferon-induced protein 35kD (IFP35) and / or N-Myc interacting protein (NMI).

28. The system according to claim 27, characterized in that: The detection device for the content and / or expression of the marker in the test sample comprises a device for detecting the content and / or expression of the marker at the gene level and / or protein level; preferably, the detection device for the content and / or expression of the marker in the test sample comprises a device for one or more detection techniques or methods selected from the group consisting of immunohistochemistry, Western blotting, Northern blotting, PCR, biochip, nucleic acid sequencing, and amino acid sequencing; preferably, the immunohistochemistry is selected from at least one of immunofluorescence, ELISA, and immunocolloidal gold. species; preferably, the test sample of the product is selected from at least one of the body fluids, tissues, cells and excreta of the subject to be tested; preferably, the body fluid comprises at least one of blood and lymph; preferably, the blood comprises at least one of serum, plasma, dried blood spots and whole blood; preferably, the tissue comprises tumor tissue; preferably, the excreta comprises at least one of urine, feces and tears; preferably, the subject to be tested comprises mammals; preferably, the subject to be tested comprises humans; preferably, the marker further comprises other markers for evaluating the efficacy of tumor immunotherapy, evaluating the prognosis of tumor immunotherapy, and / or selecting a treatment regimen.

29. The use, tumor immunotherapy synergist, anti-tumor drug composition, anti-tumor drug kit, marker combination, product, construction method, system according to any one of claims 1-17, 19-28, characterized in that: The tumor comprises a solid tumor and / or a blood tumor; Preferably, the solid tumor comprises head and neck squamous cell carcinoma, liver cancer, colorectal cancer, bladder cancer, breast cancer, cervical cancer, prostate cancer, glioma, melanoma, pancreatic cancer, nasopharyngeal cancer, lung cancer, gastric cancer, adrenal cortical carcinoma, pararenal cortical carcinoma, anal cancer, appendix cancer, astrocytoma, atypical teratoma, rhabdoid tumor, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain tumor, bronchial tumor, Burkitt's lymphoma, carcinoid tumor, heart tumor, bile duct epithelial carcinoma, chordoma, colorectal cancer, craniopharyngioma, ductal carcinoma in situ, embryonic tumor, endometrial cancer, ependymoma, esophageal cancer, olfactory neuroblastoma, intracranial germ cell tumor, extragonadal germ cell tumor, eye cancer, fallopian tube cancer, gallbladder cancer, head and neck cancer, hypopharyngeal cancer, Kaposi's sarcoma, kidney cancer, Langerhans At least one of: cell histiocytosis, laryngeal cancer, lip cancer, oral cancer, Merkel cell carcinoma, malignant mesothelioma, multiple endocrine neoplasia syndrome, mycosis fungoides, sinonasal cancer, neuroblastoma, non-small cell lung cancer, ovarian cancer, pancreatic neuroendocrine tumors, islet cell tumors, papillomatosis, paraganglioma, sinonasal cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pituitary tumor, pleuropulmonary blastoma, primary peritoneal cancer, retinoblastoma, salivary gland tumor, sarcoma, Sezary syndrome, skin cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, squamous cell carcinoma, testicular cancer, thymoma and thymic cancer, thyroid cancer, urethral cancer, uterine cancer, endometrial and uterine sarcoma, vaginal cancer, vascular tumor, vulvar cancer and single myeloma; Preferably, the blood tumor is selected from at least one of B-cell acute lymphoid leukemia, T-cell acute lymphoid leukemia, acute lymphoid leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia, B-cell prolymphocytic leukemia, blastic plasmacytoid dendritic cell tumor, Burkitt's lymphoma, diffuse large B-cell lymphoma, follicular lymphoma, hairy cell leukemia, small cell or large cell-follicular lymphoma, MALT lymphoma, mantle cell lymphoma, marginal zone lymphoma, multiple myeloma, non-Hodgkin's lymphoma, plasmablastic lymphoma, plasmacytoid dendritic cell tumor, Waldenstrom macroglobulinemia and preleukemia; Preferably, the immunotherapy comprises at least one of monoclonal antibody therapy, immune checkpoint inhibitor therapy, adoptive cell therapy, oncolytic virus therapy, and tumor vaccine therapy; Preferably, the target of the monoclonal antibody is selected from one of CD20, HER2, VEGF / VEGFR, EGFR, CD19, FGL1, CD47, CD3, CD30, CD33, CD38, CD52, αVβ3, α5β1, FAP, Tenascin, CEA, EPCAM, PSMA, GAN-GD2, GAN-GD3, GM2, and IGF-IR; Preferably, the immune checkpoint inhibitor is an inhibitor that acts on T cell negative co-stimulatory (co-inhibitory) molecules and / or their respective ligands; Preferably, the negative co-stimulatory (co-inhibitory) molecules acting on T cells and / or their respective ligands are selected from CTLA-4, PD-1, PD-L1, PD-L2, B7-1, B7-2, B7-H3, B7-H4, B7-H6, A2aR, IDO, TIM-3, BTLA, VISTA, TIGIT, LAG-3, CD40, CD20, CD96, CD73, CD160, STING, CEA, CD47, PVRIG, one or more of LAIR1, 2B4, KIR, CEACAM1, GARP, PS, CSF1R, CD94 / NKG2A, TDO, TNFR, and DcR3; Preferably, the inhibitor acting on T cell negative co-stimulatory (co-inhibitory) molecules and / or their respective ligands comprises any one of (b1) to (b3): ​​(b1) antibodies that specifically bind (neutralize) T cell negative co-stimulatory (co-inhibitory) molecules and / or their respective ligands; (b2) ligand proteins or polypeptides that specifically bind (neutralize) T cell negative co-stimulatory (co-inhibitory) molecules and / or their respective ligands; (b3) non-protein compounds that specifically bind (neutralize) T cell negative co-stimulatory (co-inhibitory) molecules and / or their respective ligands; Preferably, the adoptive cell therapy comprises at least one of chimeric antigen receptor T cell (CAR-T) therapy, chimeric antigen receptor NK cell (CAR-NK) therapy, T cell receptor chimeric T cell (TCR-T) therapy, tumor infiltrating immune cell (TILs) therapy, cytokine-induced killer (CIK) cell therapy, lymphokine-activated killer (LAK) cell therapy, and natural killer (NK) cell therapy; Preferably, the oncolytic virus comprises at least one of alphavirus, adenovirus, vaccinia virus, Sindbis virus, Seneca Valley virus, coxsackie virus, measles virus, reovirus, vaccinia virus, Newcastle disease virus, vesicular stomatitis virus, herpes simplex virus, polio virus, influenza virus, mumps virus and parvovirus; further comprises at least one of alphavirus, adenovirus, vaccinia virus, measles virus, vesicular stomatitis virus, and herpes simplex virus; Preferably, the alphavirus comprises at least one of an M1 virus and a Geta virus; Preferably, the tumor vaccine comprises at least one of a dendritic cell (DCs) vaccine, a nucleic acid vaccine and a polypeptide vaccine.

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