KR20240027528A - Biomarkers for prognosis of cervical cancer and prediction of treatment response to simultaneous chemoradiation therapy and methods for providing information on prognosis of cervical cancer using the biomarkers - Google Patents

Abstract

The present invention relates to a biomarker for predicting the prognosis of cervical cancer. When using the biomarker of the present invention, it is possible to select patients into a high-risk group, intermediate-risk group, or low-risk group, so that customized treatment for each patient can be provided according to the prognosis prediction. possible.

Description

Biomarkers for prognosis of cervical cancer and prediction of treatment response to simultaneous chemoradiation therapy and methods for providing information on prognosis of cervical cancer using the biomarkers}

The present invention relates to a biomarker for prognosis of cervical cancer and prediction of treatment response to simultaneous chemotherapy and radiation therapy and a method of providing information using the same.

Cervical cancer can refer to female reproductive cancer that occurs in the cervix, the entrance to the uterus.

Meanwhile, although the diagnosis rate of early-stage cervical cancer has increased due to the development of diagnostic techniques and screening tests, it is reported that many patients are still diagnosed at a later stage with a poor prognosis. Therefore, it may be important to receive timely and accurate diagnosis and appropriate treatment to increase the survival rate of cervical cancer patients.

When diagnosing cervical cancer, standard treatment guidelines such as surgery, radiation therapy, and chemotherapy are proposed depending on the disease stage based on the NCCN (National Comprehensive Cancer Network) and FIGO (International Federation of Gynecology and Obstetrics) stages. However, even if a patient is treated according to the above guidelines, each patient may have a different prognosis due to the heterogeneity of the tumor.

In other words, as more accuracy in prognosis prediction is required to improve medical services, biomarkers that can predict the prognosis of cervical cancer with high accuracy, including prediction of treatment response to simultaneous chemotherapy and radiation therapy, and the use of these There is a need for the development of information provision methods.

The technology behind the invention has been written to facilitate easier understanding of the invention. It should not be understood as an admission that matters described in the technology underlying the invention exist as prior art.

Meanwhile, the inventors of the present invention noted that several genes closely interact with each other during the development and progression of tumors and that individual gene expression levels may differ.

In particular, the inventors of the present invention are able to predict the prognosis of cervical cancer for different individuals and predict treatment response to simultaneous chemotherapy and radiotherapy by using biomarkers that can predict prognosis by evaluating gene expression patterns, and thus. It was recognized that tailored personal medicine was possible.

As a result, the inventors of the present invention were able to establish genes as biomarkers that can predict the prognosis of cervical cancer, including treatment response to concurrent chemotherapy and radiotherapy, and determine the treatment direction accordingly, and the clinicopathological correlation of the biomarkers. was able to confirm.

Furthermore, the inventors of the present invention provide cervical cancer prognosis prediction and information related thereto, including prediction of treatment response to simultaneous chemotherapy and radiotherapy based on the biomarkers, so that setting treatment direction for each individual can be more easily performed. It was expected that this could contribute to improving prognosis.

As a result, the inventors of the present invention have developed a biomarker for predicting the prognosis of cervical cancer according to the present invention.

Accordingly, the inventors of the present invention were able to recognize that by introducing the biomarker of the present invention, the limitations of the conventional diagnosis and information provision system could be supplemented and an efficient treatment method could be proposed accordingly.

Accordingly, the problem to be solved by the present invention is to provide a biomarker for predicting the prognosis of cervical cancer through the biomarker of the present invention and a method of providing information using the same.

The problems of the present invention are not limited to the problems mentioned above, and other problems not mentioned will be clearly understood by those skilled in the art from the description below.

In order to solve the problems described above , a biomarker for predicting the prognosis of cervical cancer, including treatment response to simultaneous chemotherapy and radiation therapy, is provided according to an embodiment of the present invention.

In one embodiment of the present invention, the biomarker may include ATP5H, SCP3, cytoplasmic pERK1/2, NANOG, and PTEN proteins or genes encoding them, but is not limited thereto.

In a specific embodiment of the present invention, the biomarker may include ATP5H, SCP, and NANOG proteins or genes encoding them, but is not limited thereto.

According to another embodiment of the present invention, the present invention provides a composition for predicting cervical cancer prognosis, including an agent for measuring the expression level of ATP5H, SCP, and NANOG proteins or genes encoding them.

In the present invention, the agent for measuring the expression level of the ATP5H, SCP, and NANOG proteins includes antibodies, oligopeptides, ligands, PNA (peptide nucleic acid), and aptamers that specifically bind to the ATP5H, SCP, and NANOG proteins. It may include one or more types selected from the group consisting of aptamer).

In the present invention, the agent for measuring the expression level of genes encoding the ATP5H, SCP, and NANOG proteins is selected from the group consisting of primers, probes, and antisense nucleotides that specifically bind to the genes encoding the ATP5H, SCP, and NANOG proteins. It may include one or more types.

According to another embodiment of the present invention, the present invention provides a kit for predicting cervical cancer prognosis, including a composition for predicting cervical cancer prognosis, including an agent for measuring the expression level of ATP5H, SCP, and NANOG proteins or genes encoding them. to provide.

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

The kit of the present invention may further include one or more other component compositions, solutions, or devices suitable for the analysis method. For example, in the present invention, the kit may further include essential elements necessary to perform a reverse transcription polymerase reaction. The reverse transcription polymerase reaction kit contains a pair of primers specific for the gene encoding the marker protein. Primers are nucleotides 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 to the nucleic acid sequence of the control gene. Other reverse transcription polymerase reaction kits include test tubes or other suitable containers, reaction buffers (pH and magnesium concentrations vary), deoxynucleotides (dNTPs), enzymes such as Taq-polymerase and reverse transcriptase, DNase, and RNase. It may include inhibitors DEPC-water, sterilized water, etc.

Additionally, the kit for predicting prognosis of the present invention may include essential elements required to perform a DNA chip. A DNA chip kit may include a substrate to which a cDNA or oligonucleotide corresponding to a gene or a fragment thereof is attached, and reagents, agents, enzymes, etc. for producing a fluorescent label probe. The substrate may also include cDNA or oligonucleotides corresponding to control genes or fragments thereof.

Additionally, the kit for predicting prognosis of the present invention may include essential elements required to perform ELISA. ELISA kits contain antibodies specific for these proteins. Antibodies are antibodies that have high specificity and affinity for a marker protein and almost no cross-reactivity to other proteins, and may be monoclonal antibodies, polyclonal antibodies, or recombinant antibodies. Additionally, ELISA kits may include antibodies specific for control proteins. Other ELISA kits include reagents that can detect bound antibodies, such as labeled secondary antibodies, chromophores, enzymes (e.g., conjugated with antibodies) and their substrates or those that can bind to antibodies. It may contain other substances, etc.

In the kit for predicting prognosis of the present invention, the fixture for the antigen-antibody binding reaction includes a nitrocellulose membrane, a PVDF membrane, a well plate synthesized from polyvinyl resin or polystyrene resin, and a glass plate. Slide glass, etc. may be used, but are not limited thereto.

In addition, the marker for the secondary antibody in the prognosis prediction kit of the present invention is preferably a conventional coloring agent that produces a color reaction, such as HRP (horseradish peroxidase), alkaline phosphatase, colloid gold, Publication Patent No. 10-2021-0050278 - 11 - Fluorescein and dyes such as FITC (poly L-lysine-fluorecein isothiocyanate) and RITC (rhodamine-B-isothiocyanate). The label may be used, but is not limited thereto.

In addition, the chromogenic substrate for inducing color development in the prognosis prediction kit of the present invention is preferably used according to the marker that produces a chromogenic reaction, including TMB (3,3',5,5'-tetramethyl bezidine), ABTS [2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)], OPD (o-phenylenediamine), etc. can be used. At this time, it is more preferable that the chromogenic substrate is provided dissolved in a buffer solution (0.1 M NaAc, pH 5.5). A chromogenic substrate such as TMB is decomposed by HRP used as a marker for the secondary antibody conjugate to produce a chromogenic deposit, and the presence or absence of the marker proteins is detected by visually checking the degree of deposition of the chromogenic deposit.

In the kit for predicting prognosis of the present invention, the washing solution preferably contains a phosphate buffer solution, NaCl, and Tween 20, and a buffer solution consisting of 0.02 M phosphate buffer solution, 0.13 M NaCl, and 0.05% Tween 20 ( PBST) is more preferred. After the antigen-antibody binding reaction, the washing solution reacts with the secondary antibody to the antigen-antibody conjugate, then adds an appropriate amount to the fixative and washes 3 to 6 times. The reaction stopping solution may preferably be a sulfuric acid solution (H ₂ SO ₄ ).

According to another embodiment of the present invention, the present invention includes the step of measuring the expression level of one or more proteins selected from the group consisting of ATP5H, SCP, and NANOG, or genes encoding the proteins, in a biological sample isolated from a subject of interest. Provides a method of providing information for predicting cervical cancer prognosis, including.

At this time, in the present invention, the method of providing information for predicting the prognosis of cervical cancer and the treatment response to simultaneous chemotherapy and radiation therapy includes the use of one or more proteins selected from the group consisting of ATP5H, SCP, and NANOG, or the gene encoding the protein. After the step of measuring the expression level, a step of classifying into any one of groups 1 to 4 according to the measured expression level may be further included.

According to the features of the present invention, the four groups classified according to the expression level of the biomarker protein are groups 1 to 4, where group 1 may be a group in which ATP5H is overexpressed (>4), and group 2 may be a group in which ATP5H is overexpressed (>4). Group 3 may be a group with low expression levels of ATP5H (≤ 4), SCP (≤ 161) and NANOG (≤ 185), and Group 4 may be a group with low expression levels of ATP5H (≤ 4), SCP (≤ 161) and NANOG (≤ 185). ) and SCP (≤ 161) proteins have low expression levels, and NANOG (>185) may be an overexpressed group.

In addition, groups 1 to 4, which are classified according to the expression level of the biomarkers, are low risk for group 1, intermediate risk for group 2 and group 3, and high risk for group 4. ) may further include a step of providing information on prognosis prediction.

In the present invention, the agent for measuring the expression level of the protein includes at least one selected from the group consisting of antibodies, oligopeptides, ligands, PNA (peptide nucleic acids), and aptamers that specifically bind to the protein. can do.

In the present invention, the expression level of the protein can be measured using protein chip analysis, immunoassay, ligand binding assay, MALDI-TOF (Matrix Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry) analysis, and SELDI-TOF (Sulface Enhanced Laser Desorption/SELDI-TOF). Ionization Time of Flight Mass Spectrometry) analysis, radioimmunoassay, radioimmunodiffusion method, Ouchteroni immunodiffusion method, rocket immunoelectrophoresis, tissue immunostaining, complement fixation assay, two-dimensional electrophoresis analysis, liquid chromatography-mass spectrometry ( It can be performed by liquid chromatography-Mass Spectrometry (LC-MS), LC-MS/MS (liquid chromatography-Mass Spectrometry/Mass Spectrometry), Western blotting, or ELISA (enzyme linked immunosorbent assay).

Additionally, in the present invention, the expression level of the protein can be measured by a multiple reaction monitoring (MRM) method.

In the present invention, in the multiple reaction monitoring method, the internal standard may be a synthetic peptide or Escherichia coli beta galactosidase in which specific amino acids constituting the target peptide are isotopically substituted.

In the present invention, the agent for measuring the expression level of the gene encoding the protein may include one or more selected from the group consisting of primers, probes, and antisense nucleotides that specifically bind to the gene encoding the protein.

In the present invention, the expression level of the gene encoding the protein can be measured using reverse transcription polymerase reaction (RT-PCR), competitive reverse transcription polymerase reaction (Competitive RT-PCR), real-time reverse transcription polymerase reaction (Real-time RTPCR), and RNase. This may be by RNase protection assay (RPA), Northern blotting, or DNA chip.

In the present invention, if the expression level of the protein or the gene encoding the protein measured in the biological sample of the subject of interest increases or decreases compared to the control group, the prognosis after treatment for cervical cancer or treatment for concurrent chemotherapy and radiation therapy It can be predicted that the reaction will be bad.

At this time, in the present invention, the method of providing information for predicting the prognosis of cervical cancer and the treatment response to simultaneous chemotherapy and radiation therapy includes the use of one or more proteins selected from the group consisting of ATP5H, SCP, and NANOG, or the gene encoding the protein. Measuring the expression level, classifying into any one of groups 1 to 4 according to the measured expression level, and then providing results for predicting cervical cancer prognosis for the individual according to the classified group. It can be included.

At this time, the step of providing results for predicting the prognosis of cervical cancer for an individual according to the classified group includes selecting one or more proteins selected from the group consisting of ATP5H, SCP, and NANOG in a biological sample isolated from the individual, or the protein. It may further include providing a cervical cancer prognosis prediction result for the individual, including the expression level of the gene encoding the individual and clinical data information of the individual. In one embodiment, the clinical data includes FIGO stage, tumor size, and lymph node metastasis. , oncological grade, and age data, but is not limited thereto.

Specific details of other embodiments are included in the detailed description and drawings.

The present invention can provide a biomarker for prognosis of cervical cancer and prediction of treatment response to simultaneous chemotherapy and radiation therapy, and a method of providing information using the same.

In particular, the present invention provides a biomarker capable of predicting the prognosis of cervical cancer, including treatment response to concurrent chemotherapy and radiation therapy, and predicts the prognosis after cervical cancer treatment, thereby providing tailored personal medicine for different individuals. can be made possible.

In other words, by utilizing a biomarker capable of predicting the prognosis of cervical cancer, the present invention can more accurately set a treatment direction for each individual and contribute to the selection of a customized treatment method.

The effects according to the present invention are not limited to the contents exemplified above, and further various effects are included in the present specification.

Figure 1 exemplarily illustrates the procedures of a method for providing information on the prognosis of cervical cancer according to an embodiment of the present invention.
Figure 2 shows the results of ranking the top 11 biomarkers related to cervical cancer recurrence using the random forest algorithm.
Figures 3 and 4 show the expression patterns of each group classified according to the expression level of the biomarkers used in various embodiments of the present invention.
Figure 5 is a diagram showing survival time (PFS) using Kaplan-Meier survival analysis.
Figure 6 shows evaluation results for prognosis prediction of biomarkers used in various embodiments of the present invention.

The advantages and features of the present invention and methods for achieving them will become clear by referring to the embodiments described in detail below along with the accompanying drawings. However, the present invention is not limited to the embodiments disclosed below and will be implemented in various different forms, but the present embodiments only serve to ensure that the disclosure of the present invention is complete and are within the scope of common knowledge in the technical field to which the present invention pertains. It is provided to fully inform those who have the scope of the invention, and the present invention is only defined by the scope of the claims. In connection with the description of the drawings, similar reference numbers may be used for similar components.

In this document, expressions such as “have,” “may have,” “includes,” or “may include” refer to the existence of the corresponding feature (e.g., a numerical value, function, operation, or component such as a part). , and does not rule out the existence of additional features.

In this document, expressions such as “A or B,” “at least one of A or/and B,” or “one or more of A or/and B” may include all possible combinations of the items listed together. . For example, “A or B,” “at least one of A and B,” or “at least one of A or B” includes (1) at least one A, (2) at least one B, Or (3) it may refer to all cases including both at least one A and at least one B.

Expressions such as “first,” “second,” “first,” or “second,” used in this document can modify various components regardless of order and/or importance, and can refer to one component. It is only used to distinguish from other components and does not limit the components. For example, a first user device and a second user device may represent different user devices regardless of order or importance. For example, the first component may be renamed as the second component without departing from the scope of rights described in this document, and similarly, the second component may also be renamed as the first component.

A component (e.g., a first component) is “(operatively or communicatively) coupled with/to” another component (e.g., a second component). When referred to as being “connected to,” it should be understood that any component may be directly connected to the other component or may be connected through another component (e.g., a third component). On the other hand, when a component (e.g., a first component) is said to be “directly connected” or “directly connected” to another component (e.g., a second component), It may be understood that no other component (e.g., a third component) exists between other components.

As used in this document, the expression “configured to” depends on the situation, for example, “suitable for,” “having the capacity to.” ," can be used interchangeably with "designed to," "adapted to," "made to," or "capable of." The term “configured (or set to)” may not necessarily mean “specifically designed to” in hardware. Instead, in some contexts, the expression “a device configured to” may mean that the device is “capable of” working with other devices or components. For example, the phrase "processor configured (or set) to perform A, B, and C" refers to a processor dedicated to performing the operations (e.g., an embedded processor), or by executing one or more software programs stored on a memory device. , may refer to a general-purpose processor (e.g., CPU or application processor) capable of performing the corresponding operations.

Terms used in this document are merely used to describe specific embodiments and may not be intended to limit the scope of other embodiments. Singular expressions may include plural expressions, unless the context clearly indicates otherwise. Terms used herein, including technical or scientific terms, may have the same meaning as generally understood by a person of ordinary skill in the technical field described in this document. Among the terms used in this document, terms defined in general dictionaries may be interpreted to have the same or similar meaning as the meaning they have in the context of related technology, and unless clearly defined in this document, have an ideal or excessively formal meaning. It is not interpreted as In some cases, even terms defined in this document cannot be interpreted to exclude embodiments of this document.

Each feature of the various embodiments of the present invention can be partially or fully combined or combined with each other, and as can be fully understood by those skilled in the art, various technical interconnections and operations are possible, and each embodiment may be implemented independently of each other. It may be possible to conduct them together due to a related relationship.

For clarity of interpretation of this specification, terms used in this specification will be defined below.

As used herein, the term “individual” may refer to any object for which the prognosis of cervical cancer is to be predicted. For example, the individual may be an individual with cervical cancer. At this time, the entity disclosed in this specification may be any mammal except human, but is not limited thereto.

As used herein, the term “biological sample” refers to any material, biological fluid, tissue, or cell obtained from or derived from an individual, including whole blood, leukocytes, and peripheral blood. Peripheral blood mononuclear cells, buffy coat, plasma, serum, sputum, tears, mucus, nasal washes, nasal intake Nasal aspirate, 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 ( It may be one or more types selected from the group consisting of synovial fluid, joint aspirate, organ secretions, cells, cell extract, and cerebrospinal fluid, and is preferably The biological sample may be cervical cancer tissue (cells) isolated from an individual.

As used herein, the term "biomarker" refers to a biological indicator that can detect changes in the body using cells, blood vessels, proteins, DNA, RNA, metabolites, etc., and the U.S. National Institutes of Health (NIH) refers to the biomarker as is defined as an indicator that can objectively measure and evaluate normal biological processes, disease progression, and drug responsiveness to treatment methods. In other words, in the case of a specific disease or cancer, it refers to a marker that can distinguish between normal and pathological conditions or predict treatment response and measure it objectively. Therefore, biomarkers must play a role in objectively measuring and evaluating normal biological processes, disease progression, and drug responsiveness to treatment methods. Target marker that confirms the presence of a drug target depending on utilization, diagnostic marker that diagnoses the presence or absence of a disease, predictive marker that can distinguish between responders and non-responders to a specific drug, and surrogate marker that can monitor the effect of drug treatment. There are markers and prognostic biomarkers that inform the prognosis of the disease, and through the biomarkers, prediction of treatment response to concurrent chemotherapy and radiation therapy for cervical cancer can be confirmed as in the present invention.

The term “prognosis” used in this specification refers to the act of predicting in advance the course of a disease and the outcome of death or survival. The above prognosis or prognosis prediction refers to the fact that the course of a disease may vary depending on the patient's physiological or environmental condition, and can be interpreted to mean all actions to predict the course of the disease before and after treatment by comprehensively considering the patient's condition. You can.

As used herein, the term “antibody” refers to a substance that specifically binds to an antigen and causes an antigen-antibody reaction. For the purposes of the present invention, antibody refers to an antibody that specifically binds to each of the ATP5H, SCP, and NANOG proteins. Antibodies of the present invention include polyclonal antibodies, monoclonal antibodies, and recombinant antibodies. The antibody can be easily produced using techniques well known in the art. For example, polyclonal antibodies can be produced by methods well known in the art, which include injecting the protein antigen into an animal and collecting blood from the animal to obtain serum containing the antibody. These polyclonal antibodies can be produced from any animal, such as goats, rabbits, sheep, monkeys, horses, pigs, cows, dogs, etc. In addition, monoclonal antibodies can be prepared using the hybridoma method (see Kohler and Milstein (1976) European Journal of Immunology 6:511-519), which is well known in the art, or phage antibody library technology (Clackson et al, Nature, 352 :624-628, 1991; Marks et al, J. Mol. Biol., 222:58, 1-597, 1991). Antibodies prepared by the above method can be separated and purified using methods such as gel electrophoresis, dialysis, salt precipitation, ion exchange chromatography, and affinity chromatography. Additionally, antibodies of the invention include intact forms with two full-length light chains and two full-length heavy chains, as well as functional fragments of the antibody molecule. A functional fragment of an antibody molecule refers to a fragment that possesses at least an antigen-binding function, and includes Fab, F(ab'), F(ab')2, and Fv.

As used herein, the term “oligopeptide” refers to a peptide consisting of 2 to 20 amino acids and may include, but is not limited to, dipeptides, tripeptides, tetrapeptides, and pentapeptides.

As used herein, the term "Peptide Nucleic Acid (PNA)" refers to an artificially synthesized polymer similar to DNA or RNA, and was first introduced by Professors Nielsen, Egholm, Berg and Buchardt at the University of Copenhagen, Denmark in 1991. . While DNA has a phosphate-ribose sugar backbone, PNA has a repeated N-(2-aminoethyl)-glycine backbone linked by peptide bonds, which greatly increases its binding force and stability to DNA or RNA, making it useful in molecular biology. , is used in diagnostic analysis and antisense therapy.

As used herein, the term “aptamer” refers to an oligonucleic acid or peptide molecule.

In the present invention, the agent for measuring the expression level of genes encoding the ATP5H, SCP, and NANOG proteins is selected from the group consisting of primers, probes, and antisense nucleotides that specifically bind to the genes encoding the ATP5H, SCP, and NANOG proteins. It may include one or more types.

As used herein, the term “primer” refers to a fragment that recognizes a target gene sequence and includes forward and reverse primer pairs, but is preferably a primer pair that provides analysis results with specificity and sensitivity. High specificity can be granted when the nucleic acid sequence of the primer is a sequence that is inconsistent with the non-target sequence present in the sample, so that the primer amplifies only the target gene sequence containing the complementary primer binding site and does not cause non-specific amplification. .

As used herein, the term "probe" refers to a substance that can specifically bind to a target substance to be detected in a sample, and a substance that can specifically confirm the presence of the target substance in the sample through said binding. it means. The type of probe is not limited as it is a material commonly used in the art, but is preferably PNA (peptide nucleic acid), LNA (locked nucleic acid), peptide, polypeptide, protein, RNA or DNA, and is most preferred. It is PNA. More specifically, the probe is a biomaterial that is derived from or similar to living organisms or includes those manufactured in vitro, such as enzymes, proteins, antibodies, microorganisms, animal and plant cells and organs, nerve cells, DNA, and It may be RNA, DNA includes cDNA, genomic DNA, and oligonucleotides, RNA includes genomic RNA, mRNA, and oligonucleotides, and examples of proteins may include antibodies, antigens, enzymes, peptides, etc.

As used herein, the term “LNA (Locked nucleic acids)” refers to a nucleic acid analog containing 2'-O, 4'-C methylene bridges. LNA nucleosides contain the common nucleic acid bases of DNA and RNA and can form base pairs according to the Watson-Crick base pairing rules. However, due to the 'locking' of the molecule due to the methylene bridge, LNA does not form the ideal shape in Watson-Crick bonding. When LNA is included in a DNA or RNA oligonucleotide, the LNA can pair with the complementary nucleotide chain more quickly and increase the stability of the double helix.

As used herein, the term "antisense" refers to a nucleotide in which an antisense oligomer hybridizes with a target sequence in RNA by Watson-Crick base pairing, typically allowing the formation of an mRNA and RNA:oligomer heteroduplex within the target sequence. It refers to an oligomer having a base sequence and a backbone between subunits. Oligomers may have exact or approximate sequence complementarity to the target sequence.

Since information about the ATP5H, SCP and NANOG proteins according to the present invention and the genes encoding them are known, those skilled in the art can easily design primers, probes or antisense nucleotides that specifically bind to the genes encoding the proteins based on this information. You can.

As used herein, the term “kit” refers to a tool that can evaluate the expression level of a biomarker by labeling a probe or antibody that specifically binds to a biomarker component with a detectable label. It includes not only direct labeling of a detectable substance related to a probe or antibody by reaction with a substrate, but also indirect labeling in which a label that develops color through reactivity with another directly labeled reagent is conjugated. It may include a chromogenic substrate solution, a washing solution, and other solutions that will undergo a color reaction with the label, and may be prepared including reagent components to be used. In the present invention, the kit may be a kit containing the essential elements required to perform RT-PCR, including a test tube, reaction buffer, deoxynucleotides (dNTPs), and Taq-polymerization, in addition to each primer pair specific for the marker gene. It may contain enzymes, reverse transcriptase, DNase, RNase inhibitor, sterile water, etc. Additionally, the kit may be a kit for detecting genes for predicting HPD prognosis that includes essential elements required to perform a DNA chip. The DNA chip kit includes a substrate to which a cDNA corresponding to a gene or a fragment thereof is attached as a probe, and the substrate may include a cDNA corresponding to a quantitative control gene or a fragment thereof. The kit of the present invention is not limited thereto, as long as it is known in the art.

As used herein, the term “individual data” refers to data that can be obtained from an individual, and may include at least one of the stage of cancer, presence of lymph node metastasis, and age.

However, it is not limited to this and may include more diverse data that can be obtained from electronic medical record data.

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. .

Evaluation 1: Derivation and performance evaluation of biomarkers

First, referring to Table 1, Table 1 shows the results of classification of the patient group for deriving the biomarker of the present invention.

To derive biomarkers for predicting cervical cancer prognosis, the patient group was divided into a learning group (70%) and a validation group (30%) as shown in Table 1, and then, as shown in Figure 2, among the biomarkers related to cervical cancer recurrence The top 11 were ranked using the random forest algorithm. Afterwards, considering the significance of the biomarkers, five major biomarkers consisting of ATP5H, SCP3, cytoplasmic pERK1/2, NANOG, and PTEN were selected.

At this time, referring to Figures 3 and 4 together, the results of dividing the risk of recurrence of cervical cancer into four groups based on the expression levels of biomarkers ATP5H, SCP3, cytoplasmic pERK1/2, NANOG, and PTEN are shown.

Group 1 was the low risk group, with ATP5H expressed above a predetermined threshold (cutoff value) (ATP5H >4). Groups 2 and 3 are intermediate risk groups. First, in group 2, it was confirmed that both ATP5H and SCP3 were expressed lower than the boundary value (ATP5H ≤ 4, SCP ≤ 161), and in group 3, ATP5H, SCP3, and NANOG were all expressed lower than the boundary value. It was confirmed that the expression level was high (ATP5H ≤ 4, SCP ≤ 161, NANOG ≤ 185), and group 4 was a high-risk group where ATP5H and SCP3 were expressed lower than the boundary value, but the expression of NANOG was higher than the boundary value (ATP5H ≤ 4, SCP ≤ 161, NANOG > 185) was confirmed.

Referring to Figure 5 together, Figure 5 is a diagram showing the survival time (PFS) using Kaplan-Meier survival analysis, in Group 1, which is a low-risk group, Groups 2 and 3, which are intermediate-risk groups, and Group 4, which is a high-risk group. It was confirmed that there was a clear difference between low-, medium-, and high-risk groups in both the learning and validation groups, indicating that the biomarker of the present invention had excellent performance in predicting cervical cancer recurrence.

Based on the above results, when using the biomarker of the present invention, it is possible to predict the prognosis for cervical cancer with high accuracy, so it is expected to contribute to increasing the life expectancy of patients by providing customized treatment for each patient.

Evaluation 2: Evaluation of biomarkers

Below, with reference to FIG. 6 and Table 2, the evaluation results of biomarkers for predicting cervical cancer prognosis according to various embodiments of the present invention will be described. Table 3 shows the results of analyzing clinicopathological characteristics by group. Figure 6 shows evaluation results for prognosis prediction of biomarkers used in various embodiments of the present invention.

Referring to Table 2, it was confirmed that the FIGO stage significantly increased from group 1 to group 4.

In group 1, a low-risk group, the proportion of subjects classified as FIGO stage IIB-IV was only 7.14%, but in group 4, a high-risk group, it was confirmed to increase to 52.24%. Tumor size also significantly increased from the low-risk group, Group 1, to the high-risk group, Group 4, and the size increased from 21.93 ± 20.31 mm to 40.25 ± 17.21 mm.

Additionally, in group 4, lymph node metastasis was confirmed at 61.54%, and the proportion of subjects with poor response to treatment was 59.62%, which was higher than that of groups 1 to 3.

In the case of LVSI, it was confirmed that the expression rate was high in groups 3 and 4 compared to other groups. Specifically, it was 80% in group 3 and 60% in group 4.

Referring further to Table 3, Table 3 is a table comparing groups 3 and 4.

As shown in Table 3, compared to group 2, group 3 individuals were younger at diagnosis, were diagnosed at a more advanced FIGO stage, had larger tumors, and had a higher LVSI expression rate.

As described above, according to the classification model of the present invention, it is possible to classify entities according to the risk of cervical cancer, develop a customized treatment plan appropriate for the patient according to tumor size, lymph node metastasis, FIGO stage, and LVSI for each group, and determine the patient's prognosis. Since it is possible to predict more accurately, it has been confirmed that it can be usefully used to improve the prognosis of cervical cancer patients.

Below, with reference to FIG. 6, evaluation results of classification models according to various embodiments of the present invention will be described.

Figure 6 shows evaluation results of classification models used in various embodiments of the present invention. Specifically, clinical information such as FIGO stage, tumor size, lymph node metastasis, oncological grade, and age were integrated into the classification model of the present invention to enable classification by combining molecular information such as expression levels of biomarkers. The C-index was compared by comparing the case considering only information and the clinical-molecular classification using the classification model of the present invention.

As a result, as shown in Figure 6, it was confirmed that cervical cancer prognosis can be predicted with higher accuracy when using a model that considers both molecular classification and clinical information compared to the case where molecular classification and clinical information are considered separately.

Although embodiments of the present invention have been described in more detail with reference to the accompanying drawings, the present invention is not necessarily limited to these embodiments, and may be modified and implemented in various ways without departing from the technical spirit of the present invention. Accordingly, the embodiments disclosed in the present invention are not intended to limit the technical idea of the present invention, but rather to explain it, and the scope of the technical idea of the present invention is not limited by these embodiments. Therefore, the embodiments described above should be understood in all respects as illustrative and not restrictive. The scope of protection of the present invention should be interpreted in accordance with the claims below, and all technical ideas within the equivalent scope should be construed as being included in the scope of rights of the present invention.

Claims (18)

Containing at least one protein or a gene encoding the same among ATP5H, NANOG, PTEN, SCP and pERK,
Biomarkers for predicting cervical cancer prognosis. According to paragraph 1,
The biomarker is,
ATP5H, SCP and NANOG Phosphorus;
Biomarkers for predicting cervical cancer prognosis. Containing an agent for measuring the expression level of ATP5H, SCP and NANOG proteins or genes encoding them,
Composition for predicting cervical cancer prognosis. According to paragraph 3,
The agent for measuring the expression level of ATP5H, SCP, and NANOG proteins consists of antibodies, oligopeptides, ligands, PNA (peptide nucleic acid), and aptamers that specifically bind to the ATP5H, SCP, and NANOG proteins. Containing one or more species selected from the group,
Composition for predicting cervical cancer prognosis. According to paragraph 3,
The agent for measuring the expression level of genes encoding proteins of ATP5H, SCP, and NANOG is one selected from the group consisting of primers, probes, and antisense nucleotides that specifically bind to genes encoding proteins of ATP5H, SCP3, and NANOG. Including the above,
Composition for predicting cervical cancer prognosis. Comprising the step of measuring the expression level of one or more proteins selected from the group consisting of ATP5H, SCP, and NANOG or genes encoding the proteins in a biological sample isolated from an individual,
Information provision method for predicting cervical cancer prognosis. According to clause 6,
Further comprising classifying into any one of groups 1 to 4 according to the measured expression level,
Information provision method for predicting cervical cancer prognosis. In clause 7,
Further comprising providing results for predicting cervical cancer prognosis for an individual according to the classified group,
Information provision method for predicting cervical cancer prognosis. According to clause 8,
In the step of providing results for predicting cervical cancer prognosis for an individual according to the classified group,
The group 1 is a group in which ATP5H is overexpressed compared to the normal control group,
Information provision method for predicting cervical cancer prognosis. According to clause 8,
In the step of providing results for predicting cervical cancer prognosis for an individual according to the classified group,
Group 2, a group with low expression of ATP5H compared to the normal control group,
Information provision method for predicting cervical cancer prognosis. According to clause 8,
In the step of providing results for predicting cervical cancer prognosis for an individual according to the classified group,
Group 3 is a group in which ATP5H, SCP, and NANOG are underexpressed,
Information provision method for predicting cervical cancer prognosis. According to clause 8,
In the step of providing results for predicting cervical cancer prognosis for an individual according to the classified group,
Group 4 is a group in which ATP5H and SCP are underexpressed and NANOG is overexpressed,
Information provision method for predicting cervical cancer prognosis. According to clause 8,
The group 1 further includes the step of classifying it as a low risk group,
Information provision method for predicting cervical cancer prognosis. According to clause 8,
Groups 2 and 3 further include the step of classifying them as intermediate risk,
Information provision method for predicting cervical cancer prognosis. According to clause 8,
Group 4 further includes the step of classifying as a high risk group,
Information provision method for predicting cervical cancer prognosis. According to clause 6,
The step of providing results for predicting cervical cancer prognosis for an individual according to the classified group,
Cervical cancer prognosis prediction results for the individual, including the expression level of one or more proteins selected from the group consisting of ATP5H, SCP, and NANOG, or the expression level of the gene encoding the protein in the biological sample isolated from the individual, and clinical data information of the individual Further comprising providing,
Information provision method for predicting cervical cancer prognosis. According to clause 16,
The clinical data are FIGO stage, tumor size, lymph node metastasis, oncological grade and age data,
Information provision method for predicting cervical cancer prognosis. A kit for predicting cervical cancer prognosis comprising the biomarker of claim 1.

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