KR102326119B1 - Biomarkers for predicting prognosis after immunotherapy of cancer - Google Patents

Abstract

The present invention relates to a biomarker for selecting patients who are predicted to develop hyperprogression (HPD) after treatment with an immunotherapy (PD-1/PD-L1). The composition of the present invention includes two selected markers (CCR7-CD45RA-, TIGIT+), and by predicting the HPD occurrence before using the immunotherapy for cancer patients, especially non-small cell lung cancer patients, immunity to non-small cell lung cancer patients In that it is possible to decide whether to use anticancer drugs or not, it is expected that life expectancy will be extended by predicting the patient group who will show a prognosis such as accelerated cancer progression after immunotherapy treatment and receiving customized treatment at an early stage.

Description

Biomarkers for predicting prognosis after immunotherapy of cancer

The present invention relates to a biomarker for predicting the prognosis after immunotherapy of cancer, and more particularly, to a biomarker for selecting patients who are predicted to have accelerated tumor progression (HPD) after immunotherapy of the cancer.

Although the diagnosis and treatment of cancer has made rapid progress in the past decade, the mortality rate due to cancer is still high. Looking at the research trends of anticancer drugs so far, the first generation can be divided into chemical anticancer drugs, the second generation targeted anticancer drugs, and the third generation immune anticancer drugs. First-generation chemical anticancer drugs have severe side effects because they damage not only cancer cells but also normal cells. That is, in order to kill cancer cells, it attacks even normal cells, destroys the patient's immune system, and due to its strong toxicity, various side effects such as hair loss, vomiting, loss of appetite, fatigue, and extreme physical strength occur. In contrast, second-generation targeted anticancer drugs have the advantage of identifying and attacking only cancer cells, but they can only be used for patients with genetic mutations, making it impossible to treat various cancers, and there are disadvantages in that treatment becomes impossible due to resistance. The third-generation immune anticancer drug has a new mechanism to kill cancer cells by activating immune cells in the body that have been suppressed, and has the advantage that it can be widely used in various carcinomas without specific genetic mutations. In recent cancer treatment, a generational shift is taking place from targeted anticancer therapy, which has been spotlighted, to immunotherapy, and in the last three to four years, cancer treatment of immunotherapy has evolved one step, and effective substances are being actively developed.

Most of the anticancer drugs currently used for anticancer treatment are drugs that target rapidly growing cancer cells. However, this is different from the second-generation anticancer agent that attacks cancer itself, and the third-generation anticancer agent, which is a therapeutic agent that induces immune cells to selectively attack only cancer cells by injecting artificial immune proteins into the body to stimulate the immune system. have. As such, immuno-cancer drugs have improved the side effects of the first-generation chemotherapy and the resistance problems of the second-generation targeted anti-cancer drugs, and have a durable response, long-term survival, and broad anti-tumor activity. ) and low toxicity profile. Immunocancer therapy is a treatment that activates the body's immune system to increase self-immunity, and through this, immune cells attack cancer cells.

However, there are concerns that immunotherapy, which has been spotlighted as an anticancer agent with fewer side effects compared to existing anticancer agents, may cause cancer to grow faster in some patients. Recently, it has been reported that hyperprogression (HPD) occurs in some patients after treatment with PD-1/PD-L1, a type of immunotherapy, but it is possible to select patients with a potential for HPD by predicting this in advance. There are currently no markers excavated. Therefore, despite the excellent effect of immuno-cancer drugs, there is a problem that leads to death more rapidly as a result of wrong use in some patients who are at risk of excessive cancer progression.

One object of the present invention is to provide a biomarker composition for predicting the presence or absence of accelerated tumor progression (HPD) especially in the prognosis after immunotherapy of cancer patients.

Another object of the present invention is to provide a kit capable of predicting the occurrence of accelerated tumor progression (HPD) even in the prognosis after immunotherapy of cancer patients.

Another object of the present invention is to provide a method of providing information for diagnosing the occurrence of accelerated tumor progression (HPD), particularly in the prognosis after immunotherapy of cancer patients.

However, the technical task to be achieved by the present invention is not limited to the tasks mentioned above, and other tasks not mentioned will be clearly understood by those of ordinary skill in the art from the following description.

Hereinafter, various embodiments described herein are described with reference to the drawings. In the following description, various specific details are set forth, such as specific forms, compositions and processes, and the like, for a thorough understanding of the present invention. However, certain embodiments may be practiced without one or more of these specific details, or in conjunction with other known methods and forms. In other instances, well-known processes and manufacturing techniques have not been described in specific detail in order not to unnecessarily obscure the present invention. Reference throughout this specification to “one embodiment” or “an embodiment” means that a particular feature, form, composition, or characteristic described in connection with the embodiment is included in one or more embodiments of the invention. Thus, references to "in one embodiment" or "an embodiment" in various places throughout this specification do not necessarily refer to the same embodiment of the invention. Additionally, the particular features, forms, compositions, or properties may be combined in any suitable manner in one or more embodiments.

Unless specifically defined in the present invention, all scientific and technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

In the present invention, "cancer immunotherapy" and "cancer immunotherapy" may be used interchangeably, and they correspond to anticancer agents having a new mechanism to kill cancer cells by activating suppressed immune cells in the body. The immuno-cancer agent has fewer side effects in that it treats the patient through strengthening the patient's own immunity, and has the effect of improving the quality of life of cancer patients and significantly extending the survival period. Immunocancer drugs exhibit anticancer effects by enhancing specificity, memory, and adaptiveness of the immune system. In other words, it uses the body's immune system to precisely attack cancer cells, so there are fewer side effects and because it uses the memory and adaptability of the immune system, patients who are effective against immune anticancer drugs can see continuous anticancer effects. As such, immune anticancer drugs improved the side effects of the first-generation chemotherapy and the resistance of the second-generation targeted anticancer drugs, and had a durable response, long-term survival, and broad anti-tumor activity. ) and low toxicity profile. Immunocancer drugs are largely divided into passive immunity and active immunotherapy. Passive immunotherapy includes immune checkpoint inhibitors, immune cell therapy, and therapeutic antibodies. Active immunotherapy includes cancer vaccines and immune-modulation agents.

In the present invention, "immune checkpoint inhibitor" refers to a drug that activates T cells and attacks cancer cells by blocking the activity of an immune checkpoint protein involved in T cell suppression. In other words, cancer cells recognize the attack of immune cells as themselves by expressing PD-L1, and there is an evasion mechanism to avoid the attack. Allow cancer cells to be eliminated. As such, new drugs developed to neutralize the immune system evasion response of cancer cells typically use antibodies that target CTLA-4, PD-1, and PD-L1 regions.

In the present invention, the "PD-1/PD-L1 inhibitor" binds to the PD-1 receptor of T cells and inhibits the immune evasion function of cancer cells. When a protein on the surface of cancer cells or hematopoietic cells called PD-L1, also called CD274 or B7-H1, is expressed on a specific cell surface, it binds to PD-1, a protein on the surface of T cells, and inhibits T cell function, resulting in cancerous cancer. Prevents cells from attacking. In the case of a PD-1 inhibitor, when the PD-1 monoclonal antibody is bound to PD-1, the suppressed T cells are activated to attack and kill cancer cells. Unlike conventional immunotherapeutic agents (cytokine therapeutics, anticancer vaccines), it binds to the binding site of cancer cells and T cells and blocks the immune evasion signal, thereby preventing the formation of an immunological synapse, thus preventing the formation of immune evasion T cells that do not interfere with immune evasion has a mechanism to destroy cancer cells. In other words, cancer cells have an immune evasion substance called PD-L1, which neutralizes immune cells and proliferates. It uses the principle that T cells that do not receive these signals kill cancer cells. In the case of PD-L1 inhibitors, they target the PD-1/PD-L1 pathway, a protein expressed on immune cells and cancer cells in the body, and block their interaction while helping immune cells to attack cancer cells. do.

In the present invention, the PD-1/PD-L1 inhibitor may be included as long as it is a PD-1/PD-L1 inhibitor approved by the US FDA or Korean FDA and used in clinical practice, for example, non-small cell lung cancer (non-small cell lung cancer). PD-1/PD-L1 inhibitors used in small cell lung carcinoma) may be included. In an embodiment of the present invention, the prognosis was confirmed in patients who used nivolumab, pembrolizumab, or atezolizumab.

In the present invention, the "biomarker" is a biological indicator that can detect changes in the body using cells, blood vessels, proteins, DNA, RNA, metabolites, etc. in the body, and the National Institutes of Health (NIH) of the United States uses the biomarker as a normal biological process. , disease progression, and drug responsiveness to treatment methods were defined as indicators that can objectively measure and evaluate. That is, in the case of a specific disease or cancer, it refers to a marker that can distinguish normal or pathological conditions or predict treatment response and objectively measure it. Therefore, biomarkers should play a role that can objectively measure and evaluate a drug's responsiveness to normal biological processes, disease progression, and treatment methods. A target marker that confirms the presence of a drug target according to the utilization, a diagnostic marker that diagnoses the presence or absence of a disease, a predictive marker that can distinguish a responder from a non-responder to a specific drug, and a surrogate marker that can monitor the effect of a drug treatment There are markers, prognostic biomarkers indicating the prognosis of a disease, and the like.

In the present invention, the term "accelerated tumor progression (hyperprogression, HPD)" is based on the size of a rapidly growing tumor, and the sum of the longest diameters (SLD) of the target lesion for one month according to the RECIST 1.1 standard. ) from the perspective of tumor growth kinetics (TGK), defined as the change in the sum of rate; TGR), and there are cases defined in terms of time to treatment failure (TTF) based on the period from tumor treatment to failure. As such, since there are various definitions, in order to more accurately determine the incidence of HPD, the inventors of the present invention initiated treatment in patients determined to have progressive disease (PD) according to RECIST 1.1 criteria in the first responsiveness assessment after PD1/PD-L1 treatment. HPD was considered to have developed based on TGK or TGR when the TGK or TGR value for the period from immediately after treatment to 12 weeks after treatment increased more than twice as compared to the value from 12 weeks before treatment to immediately before the start of treatment. and HPD based on TTF was considered to be less than 2 months. More specifically, the interpretation was limited to cases that met both the definitions based on TGK and TGR based on tumor growth dynamics.

The "prognosis" of the present invention refers to an act of predicting in advance the course of a disease and the outcome of death or survival. The prognosis or prediction of prognosis may be interpreted to mean any act of predicting the course of a disease before/after treatment by considering the patient's condition comprehensively, as the course of the disease may vary depending on the physiological or environmental condition of the patient. can For the purpose of the present invention, the prognosis is interpreted as an action to determine whether to use an immunotherapy drug by predicting in advance the occurrence of accelerated tumor progression (HPD) after immunotherapy of cancer, preferably PD-1/PD-L1 treatment can be Specifically, a poor prognosis means that after PD-1/PD-L1 treatment, a patient's cancer progression rate is accelerated, resulting in a lower survival rate and a high probability of death in the near future.

As such, the present invention accurately predicts the occurrence of accelerated tumor progression (HPD) after treatment with an immune anticancer drug (PD-1/PD-L1), so that the therapeutic effect for each individual patient can be expected in advance, and the therapeutic effect cannot be expected. Some patients are selected so that they can prolong substantial survival.

In the present invention, "tumor" or "cancer" is a disease in which the cell cycle is not regulated and continues to divide, and is divided into Carcinoma and Sarcoma depending on the site of occurrence. Carcinoma refers to a malignant tumor that arises from epithelial cells such as mucous membranes and skin, and sarcoma refers to a malignant tumor that arises from non-epithelial cells such as muscle, connective tissue, bone, cartilage, and blood vessels. The cancer is preferably breast cancer, uterine cancer, esophageal cancer, stomach cancer, brain cancer, rectal cancer, colorectal cancer, lung cancer, skin cancer, ovarian cancer, cervical cancer, kidney cancer, blood cancer, pancreatic cancer, prostate cancer, testicular cancer, laryngeal cancer, oral cancer, head and neck cancer , thyroid cancer, liver cancer, bladder cancer, osteosarcoma, lymphoma, leukemia, etc., and cancer progression such as tumor differentiation and/or proliferation is not limited as long as it is a type of cancer dependent on the cancer cells described in the present invention.

In the present invention, "non-small-cell lung cancer (NSCLC)" is a type of lung cancer and refers to non-small cell lung cancer, not small cell lung cancer, depending on the tissue type. Non-small cell lung cancer is classified into types according to the location of the cancer. There are squamous cell carcinoma that occurs in the bronchi, adenocarcinoma that occurs in the relatively small bronchial epithelium, and large cell carcinoma that occurs mainly near the lung surface. Non-small cell lung cancer is a type of carcinoma that has a relatively slow growth rate compared to small cell lung cancer, spreads to surrounding tissues, and then metastasizes later.

According to one embodiment of the present invention, CC chemokine receptor type 7 (CC chemokine receptor type 7; CCR7), tyrosine protein phosphatase receptor type C isoform (Receptor type tyrosine-protein phosphatase C isoform; CD45RA) and Ig and It relates to a biomarker composition for predicting prognosis after immunotherapy of cancer patients, comprising at least one protein of T cell immunoreceptor with Ig and ITIM domains (TIGIT) or a gene encoding the same.

In the present invention, the CCR7 may consist of the amino acid sequence represented by SEQ ID NO: 1, but is not limited thereto.

In the present invention, the CD45RA may consist of the amino acid sequence represented by SEQ ID NO: 2, but is not limited thereto.

In the present invention, the TIGIT may consist of the amino acid sequence represented by SEQ ID NO: 3, but is not limited thereto.

In a preferred embodiment of the present invention, the biomarker is a CCR7 protein or a gene encoding the same; and CD45RA protein or a gene encoding the same, but is not limited thereto.

In a preferred embodiment of the present invention, the biomarker may include a TIGIT protein or a gene encoding the same, but is not limited thereto.

In the present invention, the biomarker may be one whose expression is measured in T cells, preferably CD8+ T cells or PD-1+CD8+ T cells.

As a preferred example in the present invention, the expression of CCR7 and CD45RA may be measured for CD8+ T cells, but the present invention is not limited thereto.

As a preferred example in the present invention, the TIGIT may be one whose expression is measured in PD-1+CD8+ T cells, but is not limited thereto.

In the present invention, the immunotherapy of the cancer patient may include any one or more of passive immunotherapy and active immunotherapy.

As a preferred example in the present invention, the passive immunotherapy may be using an immune checkpoint inhibitor, an immune cell therapy, or a therapeutic antibody, but is not limited thereto.

In a preferred embodiment of the present invention, the immune checkpoint inhibitor may be an expression inhibitor of CTLA-4, PD-1 or PD-L1, preferably an anti-CTLA-4, anti-PD-1 antibody or anti-PD-L1 antibody. can

In a preferred embodiment of the present invention, the active immunotherapy may be performed using cancer treatment vaccines or immune-modulation agents, but is not limited thereto.

In addition, in the present invention, the prognosis may be the presence or absence of accelerated tumor progression (HPD) occurrence after immunotherapy of a cancer patient.

In the present invention, the cancer is breast cancer, uterine cancer, esophageal cancer, stomach cancer, brain cancer, rectal cancer, colon cancer, lung cancer, skin cancer, ovarian cancer, cervical cancer, kidney cancer, blood cancer, pancreatic cancer, prostate cancer, testicular cancer, laryngeal cancer, oral cancer, head and neck cancer , thyroid cancer, liver cancer, bladder cancer, osteosarcoma, lymphoma, leukemia, and a combination thereof may be a cancer selected from the group consisting of, preferably lung cancer.

According to another embodiment of the present invention, a protein comprising at least one of CCR7, CD45RA and TIGIT; Or it relates to a composition for predicting the prognosis after immunotherapy of cancer patients, including an agent for measuring the expression level of the gene encoding the same.

In a preferred embodiment of the present invention, the composition for predicting prognosis comprises an agent for measuring the expression level of CCR7 protein or a gene encoding the same; and an agent for measuring the expression level of the CD45RA protein or a gene encoding the same, but is not limited thereto.

In a preferred embodiment of the present invention, the composition for predicting prognosis may include an agent for measuring the expression level of the TIGIT protein or a gene encoding the same, but is not limited thereto.

The CCR7, CD45RA or TIGIT protein in the composition for predicting the prognosis of the present invention; Alternatively, the expression level of the gene encoding the same may be measured in T cells, preferably CD8+ T cells or PD-1+CD8+ T cells.

As a preferred example in the present invention, the expression level of the CCR7 and CD45RA proteins or a gene encoding each of them may be measured for CD8+ T cells, but is not limited thereto.

As a preferred example in the present invention, the expression level of the TIGIT protein or a gene encoding the same may be measured in PD-1 + CD8 + T cells, but is not limited thereto.

In the present invention, the agent for measuring the expression level of the CCR7, CD45RA or TIGIT protein is an antibody, oligopeptide, ligand, PNA (peptide nucleic acid) and aptamer ( aptamer) may include one or more selected from the group consisting of.

In the present invention, the "antibody" refers to a substance that specifically binds to an antigen and causes an antigen-antibody reaction. For the purposes of the present invention, an antibody refers to an antibody that specifically binds to each of the CCR7, CD45RA or TIGIT proteins. Antibodies of the present invention include polyclonal antibodies, monoclonal antibodies and recombinant antibodies. The antibody can be easily prepared using techniques well known in the art. For example, the polyclonal antibody can be produced by a method well known in the art, including the process of injecting an antigen of the protein into an animal and collecting blood from the animal to obtain a serum containing the antibody. Such polyclonal antibodies can be prepared from any animal such as goat, rabbit, sheep, monkey, horse, pig, cow, dog, and the like. In addition, monoclonal antibodies can be prepared using the hybridoma method well known in the art (see Kohler and Milstein (1976) European Journal of Immunology 6:511-519), or the phage antibody library technology (Clackson et al, Nature, 352). :624-628, 1991; Marks et al, J. Mol. Biol., 222:58, 1-597, 1991). The antibody prepared by the above method may be separated and purified using methods such as gel electrophoresis, dialysis, salt precipitation, ion exchange chromatography, and affinity chromatography. In addition, the antibodies of the present invention include functional fragments of antibody molecules as well as complete forms having two full-length light chains and two full-length heavy chains. A functional fragment of an antibody molecule means a fragment having at least an antigen-binding function, and includes Fab, F(ab'), F(ab')2 and Fv.

In the present invention, the "oligopeptide" is a peptide consisting of 2 to 20 amino acids and may include a dipeptide, a tripeptide, a tetrapeptide, and a pentapeptide, but is not limited thereto.

In the present invention, "PNA (Peptide Nucleic Acid)" refers to an artificially synthesized, DNA or RNA-like polymer, and was first introduced by Professors Nielsen, Egholm, Berg and Buchardt of the University of Copenhagen, Denmark in 1991. Whereas DNA has a phosphate-ribose sugar backbone, PNA has a repeated N-(2-aminoethyl)-glycine backbone linked by peptide bonds, which greatly increases binding strength and stability to DNA or RNA, resulting in molecular biology , diagnostic assays and antisense therapy. PNA is described in Nielsen PE, Egholm M, Berg RH, Buchardt O (December 1991). "Sequence-selective recognition of DNA by strand displacement with a thymine-substituted polyamide". Science 254 (5037): 1497-1500.

In the present invention, the "aptamer" is an oligonucleic acid or a peptide molecule, and the general description of the aptamer is described in Bock LC et al., Nature 355(6360):5646(1992); Hoppe-Seyler F, Butz K "Peptide aptamers: powerful new tools for molecular medicine". J Mol Med. 78(8):42630(2000); Cohen BA, Colas P, Brent R. "An artificial cell-cycle inhibitor isolated from a combinatorial library". Proc Natl Acad Sci USA. 95(24): 142727 (1998).

In the present invention, the agent for measuring the expression level of the gene encoding the CCR7, CD45RA or TIGIT protein is selected from the group consisting of a primer, a probe and an antisense nucleotide that specifically binds to the gene encoding the CCR7, CD45RA or TIGIT protein. It may include one or more types.

In the present invention, the "primer" is a fragment that recognizes a target gene sequence, and includes a pair of forward and reverse primers, but preferably, a primer pair that provides analysis results with specificity and sensitivity. High specificity can be conferred when the primer's nucleic acid sequence is a sequence that is inconsistent with the non-target sequence present in the sample, so that only the target gene sequence containing the complementary primer binding site is amplified and the primer does not cause non-specific amplification. .

In the present invention, the "probe" refers to a substance capable of specifically binding to a target substance to be detected in a sample, and refers to a substance capable of specifically confirming the presence of a target substance in a sample through the binding. The type of probe is not limited as a material commonly used in the art, but preferably PNA (peptide nucleic acid), LNA (locked nucleic acid), peptide, polypeptide, protein, RNA or DNA, and most preferably It is PNA. More specifically, the probe includes a biomaterial derived from or similar thereto or manufactured in vitro, for example, enzymes, proteins, antibodies, microorganisms, animal and plant cells and organs, neurons, DNA, and It may be RNA, DNA includes cDNA, genomic DNA, and oligonucleotides, RNA includes genomic RNA, mRNA, and oligonucleotides, and examples of proteins include antibodies, antigens, enzymes, peptides, and the like.

In the present invention, the "LNA (Locked nucleic acids)" means a nucleic acid analog including a 2'-O, 4'-C methylene bridge [J Weiler, J Hunziker and J Hall Gene Therapy (2006) 13, 496.502) ]. LNA nucleosides include common nucleic acid bases in DNA and RNA, and can form base pairs according to Watson-Crick base pairing rules. However, due to the 'locking' of the molecule due to the methylene bridge, the LNA does not form an ideal shape in the Watson-Crick bond. When LNAs are incorporated into DNA or RNA oligonucleotides, LNAs can pair with complementary nucleotide chains more rapidly, increasing the stability of the double helix. In the present invention, the "antisense" means that the antisense oligomer is hybridized with a target sequence in RNA by Watson-Crick base pairing, and typically mRNA and RNA in the target sequence: A sequence of nucleotide bases allowing the formation of an oligomeric heteroduplex. and oligomers having an inter-subunit backbone. An oligomer may have exact sequence complementarity or approximate complementarity to a target sequence.

Since the CCR7, CD45RA or TIGIT protein according to the present invention, or information on the gene encoding them, is known, those skilled in the art can easily design primers, probes or antisense nucleotides that specifically bind to the gene encoding the protein based on this information. You can do it.

In the present invention, the prognosis may be the presence or absence of accelerated tumor progression (HPD) occurrence after immunotherapy of cancer patients.

In the present invention, the cancer is breast cancer, uterine cancer, esophageal cancer, stomach cancer, brain cancer, rectal cancer, colon cancer, lung cancer, skin cancer, ovarian cancer, cervical cancer, kidney cancer, blood cancer, pancreatic cancer, prostate cancer, testicular cancer, laryngeal cancer, oral cancer, head and neck cancer , thyroid cancer, liver cancer, bladder cancer, osteosarcoma, lymphoma, leukemia, and a combination thereof may be a cancer selected from the group consisting of, preferably lung cancer.

According to another embodiment of the present invention, it relates to a kit for predicting the prognosis after immunotherapy of a cancer patient, comprising the composition for predicting the prognosis after immunotherapy of a cancer patient of the present invention.

In the present invention, "kit" refers to a tool capable of evaluating the expression level of a biomarker by labeling a probe or antibody that specifically binds to a biomarker component with a detectable label. In addition to direct labeling of a detectable substance with respect to a probe or antibody by reaction with a substrate, it includes indirect labeling in which a color-generating label is conjugated by reactivity with another directly labeled reagent. It may include a chromogenic substrate solution, a washing solution, and other solutions to undergo a color reaction with the label, and may be prepared including reagent components used. In the present invention, the kit may be a kit including essential elements necessary for performing RT-PCR, and in addition to each primer pair specific for a marker gene, a test tube, reaction buffer, deoxynucleotides (dNTPs), Taq-polymerization enzymes, reverse transcriptase, DNase, RNase inhibitors, sterile water, and the like. In addition, the kit may be a kit for detecting a gene for predicting HPD prognosis including essential elements necessary for performing a DNA chip. The DNA chip kit may include a substrate to which cDNA corresponding to a gene or fragment thereof is attached as a probe, and the substrate may include cDNA corresponding to a quantitative control gene or fragment thereof. The kit of the present invention is not limited thereto, as long as it is known in the art.

In the present invention, the kit may be an RT-PCR kit, a DNA chip kit, an ELISA kit, a protein chip kit, a rapid kit, or a multiple reaction monitoring (MRM) kit.

The kit of the present invention may further include one or more other component compositions, solutions or devices suitable for the assay method. For example, in the present invention, the kit may further include essential elements necessary for performing the reverse transcription polymerase reaction. The reverse transcription polymerase reaction kit includes a pair of primers specific for a gene encoding a marker protein. The primer is a nucleotide having a sequence specific to the nucleic acid sequence of the gene, and may have a length of about 7 bp to 50 bp, more preferably, about 10 bp to 30 bp. It may also include primers specific for the nucleic acid sequence of the control gene. Other reverse transcription polymerase reaction kits include test tubes or other suitable containers, reaction buffers (with varying pH and magnesium concentrations), deoxynucleotides (dNTPs), enzymes such as Taq-polymerase and reverse transcriptase, DNase, RNase inhibitor DEPC -Water (DEPC-water), sterile water, etc. may be included.

In addition, the kit for predicting prognosis of the present invention may include essential elements necessary for performing a DNA chip. The DNA chip kit may include a substrate to which cDNA or oligonucleotide corresponding to a gene or fragment thereof is attached, and reagents, agents, enzymes, etc. for preparing a fluorescently-labeled probe. The substrate may also contain cDNA or oligonucleotides corresponding to control genes or fragments thereof.

In addition, the kit for predicting prognosis of the present invention may include essential elements necessary for performing ELISA. The ELISA kit contains an antibody specific for this protein. Antibodies are antibodies with high specificity and affinity for a marker protein and little cross-reactivity with other proteins, and are monoclonal antibodies, polyclonal antibodies, or recombinant antibodies. The ELISA kit may also include an antibody specific for a control protein. Other ELISA kits include reagents capable of detecting bound antibody, such as labeled secondary antibodies, chromophores, enzymes (eg, conjugated with an antibody) and substrates thereof or capable of binding the antibody. other materials and the like.

In the prognosis prediction kit of the present invention, as a fixture for antigen-antibody binding reaction, a nitrocellulose membrane, a PVDF membrane, a polyvinyl resin or a polystyrene resin, a well plate synthesized from a glass, A slide glass or the like may be used, but is not limited thereto.

In addition, in the kit for predicting prognosis of the present invention, the label of the secondary antibody is preferably a conventional coloring agent that develops a color reaction, HRP (horseradish peroxidase), basic dephosphorylation enzyme (alkaline phosphatase), colloidal gold (colloid gold), Labels such as fluorescein and dye such as FITC (poly L-lysine-fluorescein isothiocyanate) and RITC (rhodamine-B-isothiocyanate) may be used, but are limited thereto it is not

In addition, the chromogenic substrate for inducing color development in the kit for predicting prognosis of the present invention is preferably used according to a color-reacting marker, TMB (3,3',5,5'-tetramethyl bezidine), ABTS[2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)], OPD(o-phenylenediamine), etc. can be used. In this case, the chromogenic substrate is more preferably provided in a dissolved state in a buffer solution (0.1 M NaAc, pH 5.5). A chromogenic substrate such as TMB is decomposed by HRP used as a label of the secondary antibody conjugate to generate chromogenic deposits, and the presence or absence of the marker proteins is detected by visually confirming the degree of deposition of the chromogenic deposits.

The washing solution in the prognosis prediction kit of the present invention preferably includes a phosphate buffer solution, NaCl, and Tween 20, and a buffer solution (PBST) consisting of 0.02 M phosphate buffer solution, 0.13 M NaCl, and 0.05% Tween 20. ) is more preferable. After the antigen-antibody binding reaction, the secondary antibody is reacted with the antigen-antibody conjugate, and then an appropriate amount is added to the immobilizer and washed 3 to 6 times. As the reaction stop solution, a sulfuric acid solution (H2SO4) may be preferably used.

According to another embodiment of the present invention, a cancer patient's immunity comprising measuring the expression level of at least one of CCR7, CD45RA, and TIGIT or a gene encoding the protein in a biological sample isolated from a subject of interest It relates to a method of providing information for predicting prognosis after treatment.

In the present invention, the biological sample refers to any material, biological fluid, tissue or cell obtained from or derived from an individual, whole blood, leukocytes, peripheral blood mononuclear cells ), buffy coat, plasma, serum, sputum, tears, mucus, nasal washes, nasal aspirate, respiration (breath), urine, semen, saliva, peritoneal washings, ascites, cystic fluid, meningeal fluid, amniotic fluid , glandular fluid, pancreatic fluid, lymph fluid, pleural fluid, nipple aspirate, bronchial aspirate, synovial fluid, joint aspirate (joint aspirate), organ secretions (organ secretions), cells (cell), cell extract (cell extract) and may be at least one selected from the group consisting of cerebrospinal fluid (cerebrospinal fluid), etc., preferably peripheral blood (peripheral blood) may be, but is not limited thereto.

In the present invention, the CCR7 may consist of the amino acid sequence represented by SEQ ID NO: 1, but is not limited thereto.

In the present invention, the CD45RA may consist of the amino acid sequence represented by SEQ ID NO: 2, but is not limited thereto.

In the present invention, the TIGIT may consist of the amino acid sequence represented by SEQ ID NO: 3, but is not limited thereto.

In a preferred embodiment of the present invention, the biomarker is a CCR7 protein or a gene encoding the same; and CD45RA protein or a gene encoding the same, but is not limited thereto.

In a preferred embodiment of the present invention, the biomarker may include a TIGIT protein or a gene encoding the same, but is not limited thereto.

In the present invention, the biomarker may be one whose expression is measured in T cells, preferably CD8+ T cells or PD-1+CD8+ T cells, in the biological sample.

As a preferred example in the present invention, the expression of CCR7 and CD45RA may be measured for CD8+ T cells in the biological sample, but is not limited thereto.

As a preferred example in the present invention, the TIGIT may be one whose expression is measured in PD-1+CD8+ T cells in the biological sample, but is not limited thereto.

In the present invention, the immunotherapy of the cancer patient may include any one or more of passive immunotherapy and active immunotherapy.

As a preferred example in the present invention, the passive immunotherapy may be using an immune checkpoint inhibitor, an immune cell therapy, or a therapeutic antibody, but is not limited thereto.

In a preferred embodiment of the present invention, the immune checkpoint inhibitor may be an expression inhibitor of CTLA-4, PD-1 or PD-L1, preferably an anti-CTLA-4, anti-PD-1 antibody or anti-PD-L1 antibody. can

In a preferred embodiment of the present invention, the active immunotherapy may be performed using cancer treatment vaccines or immune-modulation agents, but is not limited thereto.

In addition, in the present invention, the prognosis may be the presence or absence of accelerated tumor progression (HPD) occurrence after immunotherapy of a cancer patient.

In the present invention, the cancer is breast cancer, uterine cancer, esophageal cancer, stomach cancer, brain cancer, rectal cancer, colon cancer, lung cancer, skin cancer, ovarian cancer, cervical cancer, kidney cancer, blood cancer, pancreatic cancer, prostate cancer, testicular cancer, laryngeal cancer, oral cancer, head and neck cancer , thyroid cancer, liver cancer, bladder cancer, osteosarcoma, lymphoma, leukemia, and a combination thereof may be a cancer selected from the group consisting of, preferably lung cancer.

In the present invention, the agent for measuring the expression level of the CCR7, CD45RA or TIGIT protein is an antibody, oligopeptide, ligand, PNA (peptide nucleic acid) and aptamer that specifically binds to the CCR7, CD45RA or TIGIT protein. It may include one or more selected from the group consisting of.

In the information providing method of the present invention, descriptions of antibodies, oligopeptides, ligands, peptide nucleic acids (PNAs), aptamers, etc. overlap with those described above, and detailed description thereof will be omitted below in order to avoid excessive complexity of the specification.

In the present invention, the measurement of the expression level of the CCR7, CD45RA or TIGIT protein is a protein chip analysis, immunoassay, ligand binding assay, MALDI-TOF (Matrix Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry) analysis, SELDI-TOF ( Sulface Enhanced Laser Desorption/Ionization Time of Flight Mass Spectrometry (Sulface Enhanced Laser Desorption/Ionization Time of Flight Mass Spectrometry) analysis, radioimmunoassay, radioimmunodiffusion method, Oukteroni immunodiffusion method, rocket immunoelectrophoresis, tissue immunostaining, complement fixation assay, 2D electrophoresis analysis, liquid chromatography It may be performed by liquid chromatography-Mass Spectrometry (LC-MS), liquid chromatography-Mass Spectrometry/Mass Spectrometry (LC-MS/MS), Western blotting, or enzyme linked immunosorbent assay (ELISA).

In the present invention, the expression level of the CCR7, CD45RA or TIGIT protein may be measured by a multiple reaction monitoring (MRM) method.

In the present invention, in the method for monitoring multiple reactions, a synthetic peptide in which a specific amino acid constituting a target peptide is substituted with an isotope or E. coli beta galactosidase may be used as the internal standard material.

In the present invention, the target peptide of CCR7 consists of the amino acid sequence represented by SEQ ID NO: 1; The target peptide of CD45RA consists of the amino acid sequence shown in SEQ ID NO: 2; The target peptide representing the TIGIT may consist of an amino acid sequence represented by SEQ ID NO: 3.

In the present invention, the agent for measuring the expression level of the gene encoding the CCR7, CD45RA or TIGIT protein is selected from the group consisting of a primer, a probe and an antisense nucleotide that specifically binds to the gene encoding the CCR7, CD45RA or TIGIT protein. It may include one or more types.

In the information providing method of the present invention, descriptions of primers, probes, etc. are duplicated with those described above, and detailed description thereof will be omitted below in order to avoid excessive complexity of the specification.

In the present invention, the measurement of the expression level of the gene encoding the CCR7, CD45RA or TIGIT protein is a reverse transcription polymerase reaction (RT-PCR), a competitive reverse transcription polymerase reaction (Competitive RT-PCR), a real-time reverse transcription polymerase reaction (Real- time RT-PCR), RNase protection assay (RPA), Northern blotting, or DNA chip.

In the present invention, when the expression level of one or more proteins selected from the group consisting of CCR7, CD45RA and TIGIT measured with respect to the biological sample of the subject of interest or a gene encoding the same increases or decreases compared to the control group, after the cancer immunotherapy The prognosis can be predicted to be poor.

As a preferred example in the present invention, when the expression level of CCR7- cells or CD45RA- cells, preferably CCR7-CD45RA- cells measured for CD8+ T cells in the biological sample of the subject of interest, is decreased compared to the control, the It can be predicted that the prognosis after cancer immunotherapy is poor.

As a preferred example in the present invention, when the expression level of TIGIT+ cells measured for CD8+ T cells, preferably PD-1+CD8+ T cells, in the biological sample of the subject of interest increases compared to the control group, after the cancer immunotherapy The prognosis can be predicted to be poor.

In the present invention, the method for providing information may be to diagnose whether or not HPD occurs or the possibility of occurrence after immunotherapy of the subject.

In the present invention, when the expression level of one or more proteins selected from the group consisting of CCR7, CD45RA and TIGIT measured in the biological sample of the subject of interest or a gene encoding the same increases or decreases compared to the control group, after the cancer immunotherapy It can be predicted that the likelihood of developing HPD is high.

As a preferred example in the present invention, when the expression level of CCR7- cells or CD45RA- cells, preferably CCR7-CD45RA- cells measured for CD8+ T cells in the biological sample of the subject of interest, is decreased compared to the control, the It can be predicted that the likelihood of developing HPD after cancer immunotherapy is high.

As a preferred example in the present invention, when the expression level of TIGIT+ cells measured for CD8+ T cells, preferably PD-1+CD8+ T cells, in the biological sample of the subject of interest increases compared to the control group, after the cancer immunotherapy It can be predicted that the likelihood of developing HPD is high.

In the present invention, the information providing method may be to predict the responsiveness of a target individual to immunotherapy.

In the present invention, when the expression level of one or more proteins selected from the group consisting of CCR7, CD45RA and TIGIT measured with respect to the biological sample of the subject of interest or a gene encoding the same increases or decreases compared to the control group, the cancer immunotherapy It can be predicted that the responsiveness to treatment is low.

As a preferred example in the present invention, when the expression level of CCR7- cells or CD45RA- cells, preferably CCR7-CD45RA- cells measured for CD8+ T cells in the biological sample of the subject of interest, is decreased compared to the control, the It can be predicted that the therapeutic responsiveness to cancer immunotherapy is low.

As a preferred example in the present invention, when the expression level of TIGIT+ cells measured for CD8+ T cells, preferably PD-1+CD8+ T cells, in the biological sample of the subject of interest increases compared to the control group, the cancer immunotherapy It can be predicted that the responsiveness to treatment is low.

In the present invention, predicting that the therapeutic responsiveness is low may be the same as or corresponding to predicting that the likelihood of HPD occurrence after cancer immunotherapy is high.

In the present invention, the term "control group" may be a normal control group in which HPD does not occur, or a median value (average value of the corresponding patient) of a population after immunotherapy of cancer patients.

In the present invention, the cancer is breast cancer, uterine cancer, esophageal cancer, stomach cancer, brain cancer, rectal cancer, colon cancer, lung cancer, skin cancer, ovarian cancer, cervical cancer, kidney cancer, blood cancer, pancreatic cancer, prostate cancer, testicular cancer, laryngeal cancer, oral cancer, head and neck cancer , thyroid cancer, liver cancer, bladder cancer, osteosarcoma, lymphoma, leukemia, and a combination thereof may be a cancer selected from the group consisting of, preferably lung cancer.

The present invention relates to a biomarker for selecting a patient predicted to develop accelerated tumor progression (HPD) after immunotherapy for cancer. When the biomarker composition of the present invention is used, it is expected that life expectancy can be extended by predicting at an early stage a patient group that will show a prognosis such as accelerated cancer progression (HPD) after immunotherapy among cancer patients and receiving customized treatment. do.

1 is a result of TGK and TGR analysis between a reference period and an experimental period in patients who showed a PD (progressive desease) response by RECIST (solid tumor reactivity evaluation criteria) 1.1 according to an embodiment of the present invention.
2 is a result of analyzing progression-free survival (PFS) according to each response after PD-1/PD-L1 treatment according to an embodiment of the present invention.
3 is a result of analyzing overall survival (OS) according to each response after PD-1/PD-L1 treatment according to an embodiment of the present invention.
4 is a result of confirming the frequency of CCR7-/CD45RA- in CD8+ T cells, according to an embodiment of the present invention.
5 is a result confirming the frequency of TIGIT+ in PD-1+ CD8+ T cells, according to an embodiment of the present invention.
6 is a result of analyzing the expression pattern of each biomarker according to RECIST (Solid Cancer Reactivity Evaluation Criteria) 1.1 classification (PR, SD, PD, HPD) according to an embodiment of the present invention.
7 is a result of measuring PFS and OS according to the expression levels of CCR7 and CD45RA of effector/memory cells according to an embodiment of the present invention.
8 is a result of measuring PFS and OS according to the TIGIT expression level in PD-1+ CD8+ T cells according to an embodiment of the present invention.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention in more detail, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples according to the gist of the present invention. .

Example 1: Non-small-cell lung cancer (NSCLC) patient group recruitment.

1.1 Patient Recruitment

From April 2014 to November 2018, 379 patients with relapsed and/or metastatic non-small cell lung cancer (NSCLC) treated with immunotherapy (PD-1, PD-L1 inhibitors) were recruited. The PD-1/PD-L1 inhibitors used above are FDA/KFDA-approved drugs that are actually used as anticancer drugs in patients, and are three types of nivolumab, pembrolizumab, and atezolizumab. drug was used. Either pre-treatment (N=23) or post-treatment (N=17) did not receive an appropriate CT scan, or target according to Response evaluation criteria in solid tumors (RECIST) RECIST 1.1 criteria (N=49) A total of 263 patients were included in the final analysis, excluding those with no lesions or other treatments with PD-1/PD-L1 inhibitors (N=27). This study was approved as a prospective study after being reviewed by the Clinical Research Ethics Review Committee, and all patients were included in the study only if they gave their consent after hearing a sufficient explanation of the study.

Immuno-oncology (PD-1/PD-L1 inhibitor) age, gender, smoking history, Eastern Cooperative Oncology Group (ECOG) performance status, lactate dehydrogenase level, albumin level, neutrophil Various data were collected on lymphocyte-to-lymphocyte ratio, metastatic site, previous treatment history, and PD-1/PD-L1 inhibitor type. In addition, data on PD-L1 expression, EGFR driver mutation, ALK rearrangement and ROS1 rearrangement were analyzed, and treatment responses were analyzed and classified according to RECIST 1.1 criteria. The analysis results are shown in Table 1 below. Table 1 below is based on the Gustave Roussy Immune Score (GRIm-Score) prognostic score of The Royal Marsden Hospital (RMH) for better patient selection for phase 1 immunotherapy trials. It is calculated by the neutrophil to lymphocyte ratio, lactate dehydrogenase (LDH) and serum albumin concentrations.

Total
(N = 263) Non-HPD
(N = 209) HPD
(N=54) Average age (range) 63 (26-85) 64 (26-85) 59 (28-79) gender male 191 157 34 woman 72 52 20 smoking history radish 95 71 24 you 168 138 30 ECOG Performance Status ≤1 203 163 40 >1 60 46 14 LDH level ≤ Above normal value 81 65 16 > Above normal value 71 44 27 albumin level >3.5 g/dl 231 183 48 ≤3.5 g/dl 32 26 6 Neutrophil to Lymphocyte Ratio ≤3 150 121 29 3<x≤6 68 52 16 >6 45 36 9 metastatic site ≤2 134 115 19 >2 129 94 35 liver metastases radish 227 189 38 you 36 20 16 RMH score 0 41 34 7 One 52 43 9 2 53 28 25 3 6 4 2 GRIM score 0 66 55 11 One 68 39 29 2 13 11 2 3 5 4 One LIPI score 0 53 45 8 One 68 45 23 2 31 19 12 number of previous treatments ≤2 146 121 25 >2 117 88 29 most recent treatment chemotherapy 195 157 38 Targeted cancer treatment 31 21 10 no treatment experience 37 31 6 histology Adenocarcinoma 175 136 39 Squamous cell carcinoma 82 70 12 Adenosquamous carcinoma 2 One One Pleomorphic carcinoma One 0 One Sarcomatoid carcinoma 3 2 One PD-L1 expression positivity 152 127 25 voice 69 55 14 Driver mutations involved in targeted therapy EGFR mutation 31 23 8 ALK rearrangement 3 2 One ROS1 rearrangement 5 5 0 drug type PD-1 inhibitors 246 195 51 PD-L1 inhibitors 17 14 3 IgG subclass of pharmaceuticals IgG1 17 14 3 IgG4 246 195 51

Table 1 shows a comparison between the patient group with HPD and the patient group without HPD. According to the analysis of the above table, a total of 263 patients were included in the analysis, and the average age was 63 years, the majority of which were male (191/263, 72.6%) and smokers (168/263, 63.9%). [ECOG 0 or 1: 203/263 (77.2%)]. The number of previous treatments varied from patient to patient and ranged from 0 to 8 treatments. To confirm PD-L1 expression, immune tissue was performed in most patients (221/263, 84%), and more than two-thirds of the patients showed a positive pattern (152/221, 68.8%). In addition, EGFR mutations (31/263, 11.8%), ALK rearrangements (2/263, 1.1%) and ROS1 rearrangements (5/263, 1.9%) were detected only in patients with adenocarcinoma. The majority of patients received PD-1 inhibitor therapy (246/263, 93.5%), and the treatment evaluation criteria were partial response (PR), stable disease (SD), and progressive disease (PD), respectively. 52 (19.8%), 112 (42.5%), and 99 (37.6%) patients. As a result of comparing the clinical variables of non-HPD patients (PR, SD, and PD) with those of HPD patients, unlike the results of previous studies (Clin Cancer Res 2017; 23(8): 1920-1928.), the elderly aged 65 years or older No association was found between age and HPD, and unlike previous studies (Clin Cancer Res 2017; 23(15): 4242-4250.), no association was found between oncogenic EGFR mutations and HPD. HPD patients were more likely to have more metastasis sites, liver metastases, and high levels of lactate dehydrogenase (LDH), and based on these variables, the prognostic scores (RMH, GRIM, and LIPI scores between the HPD and non-HPD groups) ), it was confirmed that the HPD group was higher than the non-HPD PD group.

1.2 New definition of accelerated tumor progression (HPD) in terms of tumor dynamics

All analyzes regarding tumor growth dynamics were evaluated based on time-series CT scans. The reference period was defined as the period from 12 weeks before receiving treatment to just before the start of treatment, and the experimental period was considered as the period from immediately after the start of treatment to 12 weeks after treatment. It was defined as tumor growth kinetics (TGK) and tumor growth rate (TGR), which was defined as the change in the sum of the longest diameters (SLDs) of the target lesions during one month according to RECIST 1.1 criteria. Similarly, TGR was defined as the log-scale-corrected change in the sum of the volumes of the target lesion during one month according to the RECIST 1.1 criteria.

In this study, the HPD based on TGK or TGR in the first response assessment after PD1/PD-L1 treatment was compared with the value of the reference period in patients determined to have progressive disease (PD) by RECIST 1.1. It was considered that the value increased more than twice, and the case of HPD based on TTF was considered to be less than 2 months as suggested in previous studies.

Comparing the incidence of HPD accordingly, 55 (20.9%), 54 (20.5%) and 98 (37.3%) patients in the cohort developed HPD as defined by TGK, TGR, and TTF, respectively. The concordance rate of each criterion was high in the results based on TGK and TGR, confirming that the definitions based on TGK and TGR tumor growth dynamics are interchangeable and can be used more universally than those based on TTF ( see Fig. 1).

1.3 Statistical analysis

The program used for statistical analysis was GraphPad Prism 6.0 version of GraphPad Software (La Jolla, CA, USA), and in order to compare the patient characteristics between the two groups, statistics were presented as t-test results. A logistic regression model was used to analyze the effects of HPD occurrence, the Kaplan-Meier method was used to represent the survival distribution, and a log rank test was used for comparison. PFS (progression-free survival) and OS (overall survival) risk ratios were analyzed through Cox regression, and p value <0.05 was considered if significant correlation was observed.

Example 2: Confirmation of the relationship between the occurrence of HPD after treatment and the survival period

In order to discover markers that can predict the occurrence of HPD, the present inventors classified and analyzed the responses after treatment with PD-1/PD-L1 into four categories. According to the RECIST version 1.1 post-treatment response evaluation criteria, partial response (PR), stable disease (SD), and progressive disease (PD) were broadly classified. Progression without tumor growth and progression with tumor growth were divided into four categories. PR is a case in which tumor progression is reduced after treatment compared to before receiving chemotherapy, which corresponds to a case where there is a therapeutic effect. HPD refers to cases of accelerated tumor progression during PD.

2.1. Analysis of progression-free survival (PFS) according to response after PD-1/PD-L1 treatment

The results of PFS analysis for patients who showed different treatment responses into the four classifications are shown in FIG. 2 (p value < 0.001). It was confirmed that the median PFS of PD patients without HPD was 48 days, whereas those of PD patients who developed HPD showed 19 days.

2.2. Analysis of overall survival (OS) according to response after PD-1/PD-L1 treatment

The results of OS analysis in the same manner as described above are shown in FIG. 3 (p value < 0.001), and it can be seen that a pattern similar to the results of PFS is shown. It was confirmed that the median OS of PD patients without HPD was 205 days, whereas those of PD patients who developed HPD showed 50 days.

PFS refers to the duration after diagnosis that the disease has not worsened, i.e., there is no progression of the disease, OS refers to the total survival time to death after diagnosis or treatment of the disease, and usually a new treatment is available for each patient. It is used to evaluate how it works. Summarizing the above results, it can be confirmed that, in the case of PD patients who develop HPD, the survival period becomes very short, and the opportunity for additional treatment is lost. Therefore, it is important to select patients who are expected to develop HPD prior to treatment to receive more appropriate treatment, and it has been confirmed that this is a task directly related to the prolongation of the survival period of cancer patients.

Example 3: Selection of biomarkers

3.1 Examination of the patient's peripheral blood

To identify potential biomarkers that could predict the development of accelerated tumor progression (HPD), we focused on CD8 T lymphocytes considering their major role in antitumor immunity, and additionally CD8 T lymphocytes from peripheral blood For classification, we analyzed the frequency of effector/memory cells using two well-known markers, CCR7 and CD45RA. Also, considering that PD-1+ CD8+ T lymphocytes from peripheral blood are tumor-reactive cells, the frequency of PD-1+ cells in CD8+ T lymphocytes was analyzed. To further elucidate the depletion state of PD-1+ CD8+ T cells based on different immune checkpoints, we calculated the frequency of TIGIT+ (severely exhausted tumor-reactive CD8+ T cells) in PD-1+ CD8+ T cells. analyzed.

3.2 Screening of biomarkers associated with accelerated tumor progression (HPD)

As a result of examining the frequency of each marker in the blood sample, the frequency of the PD-1+ marker in CD8+ T cells did not show any difference between the group with and without accelerated tumor progression (HPD). On the other hand, the CCR7- and CD45RA- and TIGIT+ markers showed differences between the patients with and without HPD. Therefore, through this, it was possible to specify a marker that can select the PD patient group with accelerated tumor progression (HPD), which is different from the general disease progression (PR/SD) patient group and the PD patient group without HPD. . Through this, it was confirmed that the combination of the biomarkers can be used to predict the prognosis of accelerated tumor progression (HPD).

Example 4: Analysis of expression patterns in patients with HPD after PD-1/PD-L1 treatment

The sample of non-small cell lung cancer (NSCLC) patients in this study to analyze the expression pattern of biomarkers is the second-largest sample among similar studies and corresponds to the first study to record the incidence of HPD in Asian non-small cell lung cancer patients. As a biomarker selected in Example 3, the results of analyzing CCR7-/CD45RA- expression in CD8+ T cells are shown in FIG. 4 . In CD8+ T cells, it was confirmed that the frequency of the effector/memory subtype (CCR7- CD45RA-) was higher in patients without HPD than in patients with HPD (see FIG. 4 ). As a biomarker selected in Example 3, the results of analyzing TIGIT+ expression in PD-1+/CD8+ T cells are shown in FIG. 5 . The high TIGIT+ frequency in heavily depleted tumor-reactive CD8+ T cells means that HPD can be predicted to develop (see FIG. 5 ).

The results of analyzing the expression patterns of each biomarker according to the four classifications (PR, SD, PD, HPD) are shown in FIG. 6 . It can be seen that the expression patterns of PD without progression of HPD, PR, and SD were identical, and that only PD with progression of HPD showed different expression patterns. The survival period (PFS/OS) according to the expression pattern of each marker was analyzed to compare. As a result, when the frequency of the effector/memory subtype (CCR7-CD45RA-) was low, both PFS and OS showed a short duration, which could be seen as the occurrence of accelerated tumor progression (HPD) (see Fig. 7). there was. On the other hand, the higher the TIGIT+ frequency, the shorter the duration of both PFS and OS, which can be seen as the occurrence of HPD (see FIG. 8). As a result of checking the expression frequency of each marker, it can be seen that these two biomarkers independently predict the prognosis in terms of PFS and OS (see FIGS. 7 and 8 ).

Based on the above results, the accelerated expression of the marker through a lower frequency of effector/memory subtypes (CCR7- CD45RA-) in CD8+ T cells and a higher frequency of marker expression (TIGIT+) in the depleted PD1+ CD8+ T cells of tumor reactivity. It is expected to contribute to increasing the life expectancy of patients by predicting patients with a high probability of developing tumor progression (HPD) in advance and deciding whether to treat them with immunotherapy (PD-1/PD-L1).

As described above in detail a specific part of the present invention, for those of ordinary skill in the art, this specific description is only a preferred embodiment, and it is clear that the scope of the present invention is not limited thereto. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

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Val Ser Ile Ser His Asn Ser 180 185 190 Cys Thr Ala Pro Asp Lys Thr Leu Ile Leu Asp Val Pro Gly Val 195 200 205 Glu Lys Phe Gln Leu His Asp Cys Thr Gln Val Glu Lys Ala Asp Thr 210 215 220 Thr Ile Cys Leu Lys Trp Lys Asn Ile Glu Thr Phe Thr Cys Asp Thr 225 230 235 240 Gln Asn Ile Thr Tyr Arg Phe Gln Cys Gly Asn Met Ile Phe Asp Asn 245 250 255 Lys Glu Ile Lys Leu Glu Asn Leu Glu Pro Glu His Glu Tyr Lys Cys 260 265 270 Asp Ser Glu Ile Leu Tyr Asn Asn His Lys Phe Thr Asn Ala Ser Lys 275 280 285 Ile Ile Lys Thr Asp Phe Gly Ser Pro Gly Glu Pro Gln Ile Ile Phe 290 295 300 Cys Arg Ser Glu Ala Ala His Gin Gly Val Ile Thr Trp Asn Pro Pro 305 310 315 320 Gln Arg Ser Phe His Asn Phe Thr Leu Cys Tyr Ile Lys Glu Thr Glu 325 330 335 Lys Asp Cys Leu Asn Leu Asp Lys Asn Leu Ile Lys Tyr Asp Leu Gln 340 345 350 Asn Leu Lys Pro Tyr Thr Lys Tyr Val Leu Ser Leu His Ala Tyr Ile 355 360 365 Ile Ala Lys Val Gln Arg Asn Gly Ser Ala Ala Met Cys His Phe Thr 370 375 380 Thr Lys Ser Ala Pro Ser Gln Val Trp Asn Met Thr Val Ser Met 385 390 395 400 Thr Ser Asp Asn Ser Met His Val Lys Cys Arg Pro Pro Arg Asp Arg 405 410 415 Asn Gly Pro His Glu Arg Tyr His Leu Glu Val Glu Ala Gly Asn Thr 420 425 430 Leu Val Arg Asn Glu Ser His Lys Asn Cys Asp Phe Arg Val Lys Asp 435 440 445 Leu Gln Tyr Ser Thr Asp Tyr Thr Phe Lys Ala Tyr Phe His Asn Gly 450 455 460 Asp Tyr Pro Gly Glu Pro Phe Ile Leu His His Ser Thr Ser Tyr Asn 465 470 475 480 Ser Lys Ala Leu Ile Ala Phe Leu Ala Phe Leu Ile Ile Val Thr Ser 485 490 495 Ile Ala Leu Leu Val Val Leu Tyr Lys Ile Tyr Asp Leu His Lys Lys 500 505 510 Arg Ser Cys Asn Leu Asp Glu Gln Gln Glu Leu Val Glu Arg Asp Asp 515 520 525 Glu Lys Gln Leu Met Asn Val Glu Pro Ile His Ala Asp Ile Leu Leu 530 535 540 Glu Thr Tyr Lys Arg Lys Ile Ala Asp Glu Gly Arg Leu Phe Leu Ala 545 550 555 560 Glu Phe Gln Ser Ile Pro Arg Val Phe Ser Lys Phe Pro Ile Lys Glu 565 570 575 Ala Arg Lys Pro Phe Asn Gln Asn Lys Asn Arg Tyr Val Asp Ile Leu 580 585 590 Pro Tyr Asp Tyr Asn Arg Val Glu Leu Ser Glu Ile Asn Gly Asp Ala 595 600 605 Gly Ser Asn Tyr Ile Asn Ala Ser Tyr Ile Asp Gly Phe Lys Glu Pro 610 615 620 Arg Lys Tyr Ile Ala Ala Gln Gly Pro Arg Asp Glu Thr Val Asp Asp 625 630 635 640 Phe Trp Arg Met Ile Trp Glu Gln Lys Ala Thr Val Ile Val Met Val 645 650 655 Thr Arg Cys Glu Glu Gly Asn Arg Asn Lys Cys Ala Glu Tyr Trp Pro 660 665 670 Ser Met Glu Glu Gly Thr Arg Ala Phe Gly Asp Val Val Val Lys Ile 675 680 685 Asn Gln His Lys Arg Cys Pro Asp Tyr Ile Ile Gln Lys Leu Asn Ile 690 695 700 Val Asn Lys Lys Glu Lys Ala Thr Gly Arg Glu Val Thr His Ile Gln 705 710 715 720 Phe Thr Ser Trp Pro Asp His Gly Val Pro Glu Asp Pro His Leu Leu 725 730 735 Leu Lys Leu Arg Arg Arg Val Asn Ala Phe Ser Asn Phe Phe Ser Gly 740 745 750 Pro Ile Val Val His Cys Ser Ala Gly Val Gly Arg Thr Gly Thr Tyr 755 760 765 Ile Gly Ile Asp Ala Met Leu Glu Gly Leu Glu Ala Glu Asn Lys Val 770 775 780 Asp Val Tyr Gly Tyr Val Val Lys Leu Arg Arg Gln Arg Cys Leu Met 785 790 795 800 Val Gln Val Glu Ala Gln Tyr Ile Leu Ile His Gln Ala Leu Val Glu 805 810 815 Tyr Asn Gln Phe Gly Glu Thr Glu Val Asn Leu Ser Glu Leu His Pro 820 825 830 Tyr Leu His Asn Met Lys Lys Arg Asp Pro Pro Ser Glu Pro Ser Pro 835 840 845 Leu Glu Ala Glu Phe Gln Arg Leu Pro Ser Tyr Arg Ser Trp Arg Thr 850 855 860 Gln His Ile Gly Asn Gln Glu Glu Asn Lys Ser Lys Asn Arg Asn Ser 865 870 875 880 Asn Val Ile Pro Tyr Asp Tyr Asn Arg Val Pro Leu Lys His Glu Leu 885 890 895 Glu Met Ser Lys Glu Ser Glu His Asp Ser Asp Glu Ser Ser Asp Asp 900 905 910 Asp Ser Asp Ser Glu Glu Pro Ser Lys Tyr Ile Asn Ala Ser Phe Ile 915 920 925 Met Ser Tyr Trp Lys Pro Glu Val Met Ile Ala Ala Gln Gly Pro Leu 930 935 940 Lys Glu Thr Ile Gly Asp Phe Trp Gln Met Ile Phe Gln Arg Lys Val 945 950 955 960 Lys Val Ile Val Met Leu Thr Glu Leu Lys His Gly Asp Gln Glu Ile 965 970 975 Cys Ala Gln Tyr Trp Gly Glu Gly Lys Gln Thr Tyr Gly Asp Ile Glu 980 985 990 Val Asp Leu Lys Asp Thr Asp Lys Ser Ser Thr Tyr Thr Leu Arg Val 995 1000 1005 Phe Glu Leu Arg His Ser Lys Arg Lys Asp Ser Arg Thr Val Tyr Gln 1010 1015 1020 Tyr Gln Tyr Thr Asn Trp Ser Val Glu Gln Leu Pro Ala Glu Pro Lys 1025 1030 1035 1040 Glu Leu Ile Ser Met Ile Gln Val Val Lys Gln Lys Leu Pro Gln Lys 1045 1050 1055 Asn Ser Ser Glu Gly Asn Lys His His Lys Ser Thr Pro Leu Leu Ile 1060 1065 1070 His Cys Arg Asp Gly Ser Gln Gln Thr Gly Ile Phe Cys Ala Leu Leu 1075 1080 1085 Asn Leu Leu Glu Ser Ala Glu Thr Glu Glu Val Val Asp Ile Phe Gln 1090 1095 1100 Val Val Lys Ala Leu Arg Lys Ala Arg Pro Gly Met Val Ser Thr Phe 1105 1110 1115 1120 Glu Gln Tyr Gln Phe Leu Tyr Asp Val Ile Ala Ser Thr Tyr Pro Ala 1125 1130 1135 Gln Asn Gly Gln Val Lys Lys Asn Asn His Gln Glu Asp Lys Ile Glu 1140 1145 1150 Phe Asp Asn Glu Val Asp Lys Val Lys Gln Asp Ala Asn Cys Val Asn 1155 1160 1165 Pro Leu Gly Ala Pro Glu Lys Leu Pro Glu Ala Lys Glu Gln Ala Glu 1170 1175 1180 Gly Ser Glu Pro Thr Ser Gly Thr Glu Gly Pro Glu His Ser Val Asn 1185 1190 1195 1200 Gly Pro Ala Ser Pro Ala Leu Asn Gln Gly Ser 1205 1210 <210> 3 <211> 244 <212> PRT <213> Homo sapiens <400> 3 Met Arg Trp Cys Leu Leu Leu Ile Trp Ala Gln Gly Leu Arg Gln Ala 1 5 10 15 Pro Leu Ala Ser Gly Met Met Thr Gly Thr Ile Glu Thr Thr Gly Asn 20 25 30 Ile Ser Ala Glu Lys Gly Gly Ser Ile Ile Leu Gln Cys His Leu Ser 35 40 45 Ser Thr Thr Ala Gln Val Thr Gln Val Asn Trp Glu Gln Gln Asp Gln 50 55 60 Leu Leu Ala Ile Cys Asn Ala Asp Leu Gly Trp His Ile Ser Pro Ser 65 70 75 80 Phe Lys Asp Arg Val Ala Pro Gly Pro Gly Leu Gly Leu Thr Leu Gln 85 90 95 Ser Leu Thr Val Asn Asp Thr Gly Glu Tyr Phe Cys Ile Tyr His Thr 100 105 110 Tyr Pro Asp Gly Thr Tyr Thr Gly Arg Ile Phe Leu Glu Val Leu Glu 115 120 125 Ser Ser Val Ala Glu His Gly Ala Arg Phe Gln Ile Pro Leu Leu Gly 130 135 140 Ala Met Ala Ala Thr Leu Val Val Ile Cys Thr Ala Val Ile Val Val 145 150 155 160 Val Ala Leu Thr Arg Lys Lys Lys Ala Leu Arg Ile His Ser Val Glu 165 170 175 Gly Asp Leu Arg Arg Lys Ser Ala Gly Gln Glu Glu Trp Ser Pro Ser 180 185 190 Ala Pro Ser Pro Pro Gly Ser Cys Val Gln Ala Glu Ala Ala Pro Ala 195 200 205 Gly Leu Cys Gly Glu Gln Arg Gly Glu Asp Cys Ala Glu Leu His Asp 210 215 220 Tyr Phe Asn Val Leu Ser Tyr Arg Ser Leu Gly Asn Cys Ser Phe Phe 225 230 235 240 Thr Glu Thr Gly

Claims (24)

delete delete delete delete delete delete delete at least one protein selected from the group consisting of CCR7, CD45RA and TIGIT; Or as a composition for predicting the prognosis after lung cancer immunotherapy, comprising an agent for measuring the expression level of the gene encoding it,
the CCR7 protein or a gene encoding the same; and CD45RA protein or a gene encoding the same is to measure the expression level in CD8+ T cells,
The TIGIT protein or the gene encoding it is to measure the expression level in PD-1 + CD8 + T cells,
The immunotherapy is using a PD-1 inhibitor or a PD-L1 inhibitor,
The prognosis prediction is accelerated tumor progression (hyperprogression; HPD) to predict the presence or absence or survival period, the composition. delete delete delete delete delete 9. The method of claim 8,
The agent for measuring the expression level of one or more proteins selected from the group consisting of CCR7, CD45RA and TIGIT is an antibody, oligopeptide, and ligand that specifically binds to one or more proteins selected from the group consisting of CCR7, CD45RA and TIGIT. , A composition comprising at least one selected from the group consisting of peptide nucleic acid (PNA) and aptamer. 9. The method of claim 8,
The agent for measuring the expression level of a gene encoding one or more proteins selected from the group consisting of CCR7, CD45RA and TIGIT is specific to a gene encoding one or more proteins selected from the group consisting of CCR7, CD45RA and TIGIT. A composition comprising at least one selected from the group consisting of a binding primer, a probe, and an antisense nucleotide. delete A kit for predicting prognosis after immunotherapy for lung cancer, comprising the composition for predicting prognosis after immunotherapy for lung cancer according to any one of claims 8, 14 and 15,
The immunotherapy is using a PD-1 inhibitor or a PD-L1 inhibitor,
The prognostic prediction is for predicting the presence or absence or survival period of accelerated tumor progression (hyperprogression; HPD), the kit. Information for predicting prognosis after immunotherapy for lung cancer, comprising measuring the expression level of one or more proteins selected from the group consisting of CCR7, CD45RA and TIGIT or a gene encoding the protein in a biological sample isolated from a subject of interest A method of providing, comprising:
the CCR7 protein or a gene encoding the same; and CD45RA protein or a gene encoding the same is to measure the expression level in CD8+ T cells,
The TIGIT protein or the gene encoding it is to measure the expression level in PD-1 + CD8 + T cells,
The immunotherapy is using a PD-1 inhibitor or a PD-L1 inhibitor,
The prognostic prediction is for predicting the presence or absence or survival period of accelerated tumor progression (hyperprogression; HPD). 19. The method of claim 18,
The step of measuring the expression level is CCR7 protein or a gene encoding it; and measuring the expression level of the CD45RA protein or a gene encoding the same. delete 19. The method of claim 18,
The step of measuring the expression level is to measure the expression level of the TIGIT protein or a gene encoding it, the method. delete 19. The method of claim 18,
the expression level of CCR7- cells, CD45RA- cells or CCR7-CD45RA- cells measured for CD8+ T cells in the biological sample of the subject of interest is decreased compared to the control;
When the expression level of TIGIT+ cells measured for PD-1+CD8+ T cells in the biological sample of the subject of interest is increased compared to the control group;
A method of predicting a high likelihood of developing accelerated tumor progression following treatment with a PD-1 inhibitor or a PD-L1 inhibitor. delete

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