Biomarker for predicting asthma treatment responsiveness to biological agent using blood proteome and use thereof

Abstract

The present invention relates to: a biomarker for predicting asthma treatment responsiveness to a biological agent, using blood proteome; and use thereof and, in particular, to: a method for predictin

Description

Biomarkers for predicting asthma treatment responsiveness to biological agents using blood proteome and their use

[0001] The present invention relates to a biomarker for predicting asthma treatment response to a biological agent using blood protein and its use, and more specifically, to a method for predicting whether a biological agent is effective in treatment or monitoring the effect of treatment by confirming the amount of a protein biomarker contained in a blood protein.

[0002] This invention claims priority to Republic of Korea Patent Application No. 10-2023-0099044, filed July 28, 2023, the entire disclosure of which is incorporated herein by reference.

[0003] Asthma is a disease characterized by bronchial hyperresponsiveness to various stimuli, causing chronic airway inflammation and clinical symptoms such as wheezing, dyspnea, and coughing caused by widespread airway narrowing. These symptoms can improve spontaneously or with treatment. In general, asthma is recognized as a chronic inflammatory disease that occurs when inflammatory cells proliferate, differentiate, and are activated by interleukin-4, 5, and 13 produced by TH2 type immune cells, and migrate and infiltrate the airways and surrounding tissues. In this case, activated inflammatory cells such as eosinophils, mast cells, and alveolar macrophages play an important role in the process of strong bronchoconstriction by secreting various inflammatory mediators (cysteine ​​leukotrienes, prostaglandins, etc.).

[0004] Because this type of asthma is typically poorly controlled and prone to recurrent exacerbations, systemic steroids are frequently used to treat exacerbated asthma. However, due to the significant side effects of frequent steroid use, biologic agents have recently emerged as new, effective treatments for asthma. Examples of biologic agents include omalizumab, mepolizumab, reslizumab, benralizumab, and dupilumab.

[0005] However, these biologics are often not covered by insurance. Even when covered, they are often used only after the first-line treatment has been completed and the disease has progressed significantly, making them cost-prohibitive. Furthermore, because biologics target different substances depending on their type, their effectiveness varies depending on the specific patient. Therefore, when attempting to treat asthma with biologics, quickly predicting therapeutic efficacy and deciding whether to proceed with treatment is crucial for reducing both financial burden and patient suffering.

[0006] Therefore, if there is a method to predict the therapeutic effect of biological agents in asthma patients, it is expected that not only will the treatment cost and suffering of severe asthma patients be significantly reduced, but the treatment effect will also be significantly increased by selecting the biological agent that is right for the individual.

[0007] The present invention has been devised to solve the problems of the prior art as described above, and comprises the steps of: a) measuring the amount of a protein contained in a biological sample obtained from an asthma patient administered a biological agent or the expression level of a gene encoding the protein; And b) a step of comparing the amount of the measured protein or the expression level of the gene encoding the protein with the amount of the protein or the expression level of the gene encoding the protein of a control group, wherein the protein is a protein expressed from one or more genes selected from the group consisting of ACADL, APOC4, CCDC158, CDHR5, CLIP1, COL1A1, CTCFL, DNAI3, FAM98C, FGD2, GPT2, H2BK1, MDN1, MS4A1, NBL1, PIKFYVE, ROCK2, SRRM1, BPIFB6, EIF3L, FBXL19, KNG1, PRG2, TPI1, and UMOD, and the control group is a biological sample obtained from the asthma patient before the administration of the biological agent. The purpose of the present invention is to provide a method for providing information on predicting the treatment responsiveness of an asthma patient to a biological agent, comprising:

[0008] In addition, the present invention aims to provide a composition for predicting treatment response to a biological agent of an asthma patient, the composition comprising as an active ingredient an agent for measuring the amount of a protein expressed from one or more genes selected from the group consisting of ACADL, APOC4, CCDC158, CDHR5, CLIP1, COL1A1, CTCFL, DNAI3, FAM98C, FGD2, GPT2, H2BK1, MDN1, MS4A1, NBL1, PIKFYVE, ROCK2, SRRM1, BPIFB6, EIF3L, FBXL19, KNG1, PRG2, TPI1, and UMOD, or the expression amount of a gene encoding the protein.

[0009] In addition, the present invention aims to provide a kit for predicting the therapeutic response of an asthma patient to a biological agent, the kit comprising a composition for predicting the therapeutic response of an asthma patient to a biological agent, and an instruction manual.

[0010] In addition, the present invention aims to provide a diagnostic device for predicting the therapeutic response of an asthma patient to a biological agent, which comprises a composition for predicting the therapeutic response of an asthma patient to a biological agent as an active ingredient.

[0011] In addition, the present invention provides a method for treating asthma, comprising the steps of: a) administering a pharmaceutically effective amount of a biological agent to an asthma patient; b) obtaining a biological sample from the asthma patient; c) measuring the amount of a protein expressed from one or more genes selected from the group consisting of BPIFB6, EIF3L, FBXL19, KNG1, PRG2, TPI1, and UMOD contained in the biological sample or the expression level of the gene; d) comparing the measured amount of the protein or the expression level of the gene with the amount of the same protein or the expression level of the gene in a control group; and e) when the measured amount of the protein or the expression level of the gene is significantly increased compared to the amount in the control group, administering the pharmaceutically effective amount of the biological agent to the patient again.

[0012] However, the technical problems to be solved by the present invention are not limited to the problems mentioned above, and other problems not mentioned can be clearly understood by a person having ordinary skill in the technical field to which the present invention belongs from the description below.

[0013] The present invention comprises the steps of: a) measuring the amount of a protein contained in a biological sample obtained from an asthma patient administered a biological agent or the expression level of a gene encoding the protein; And b) a step of comparing the amount of the measured protein or the expression level of the gene encoding the protein with the amount of the protein or the expression level of the gene encoding the protein of a control group, wherein the protein is a protein expressed from one or more genes selected from the group consisting of ACADL, APOC4, CCDC158, CDHR5, CLIP1, COL1A1, CTCFL, DNAI3, FAM98C, FGD2, GPT2, H2BK1, MDN1, MS4A1, NBL1, PIKFYVE, ROCK2, SRRM1, BPIFB6, EIF3L, FBXL19, KNG1, PRG2, TPI1, and UMOD, and the control group is a blood sample obtained from the asthma patient before the administration of the biological agent. A method for providing information on predicting the treatment responsiveness of an asthma patient to a biological agent is provided.

[0014] In one specific embodiment of the present invention, the method may further include a step of classifying the therapeutic response to the biological agent as poor when the amount of a protein expressed from one or more genes selected from the group consisting of c) ACADL, APOC4, CCDC158, CDHR5, CLIP1, COL1A1, CTCFL, DNAI3, FAM98C, FGD2, GPT2, H2BK1, MDN1, MS4A1, NBL1, PIKFYVE, ROCK2, and SRRM1 or the expression level of the genes is significantly increased compared to the control group.

[0015] In another specific embodiment of the present invention, the method may further include the step of classifying the patient as exhibiting a therapeutic effect for the biological agent when the amount of protein expressed from one or more genes selected from the group consisting of c) BPIFB6, EIF3L, FBXL19, KNG1, PRG2, TPI1, and UMOD or the expression level of the genes is significantly increased compared to the control group.

[0016] In another specific embodiment of the present invention, the biological sample is preferably blood, plasma, or serum, but is not limited thereto as long as it is a sample that can be obtained non-invasively and contains proteins or mRNA.

[0017] In another specific embodiment of the present invention, the biological agent may be at least one selected from the group consisting of omalizumab, mepolizumab, reslizumab, benralizumab, and dupilumab, and is not limited thereto as long as it is a biological agent used as a treatment for asthma.

[0018] In another specific embodiment of the present invention, the step of measuring the amount of the protein or the expression level of the gene encoding the protein may be performed by Western blotting, enzyme linked immunosorbent assay (ELISA), radioimmunoassay, radioimmunodiffusion, Ouchterlony immunodiffusion, Rocket immunoelectrophoresis, immunohistochemical staining, immunoprecipitation assay, complete fixation assay, flow cytometry (FACS), protein chip, gas chromatography, liquid chromatography, mass spectrometry, measurement using affibody, proximity extension assay, reverse transcription polymerase chain reaction (RT-PCR), competitive RT-PCR, real-time It can be measured by one or more methods selected from the group consisting of real-time RT-PCR, RNase protection assay, Northern blotting, and DNA microarray chip.

[0019] In addition, the present invention provides a composition for predicting treatment response to a biological agent of an asthma patient, comprising as an active ingredient an agent that measures the amount of a protein expressed from one or more genes selected from the group consisting of ACADL, APOC4, CCDC158, CDHR5, CLIP1, COL1A1, CTCFL, DNAI3, FAM98C, FGD2, GPT2, H2BK1, MDN1, MS4A1, NBL1, PIKFYVE, ROCK2, SRRM1, BPIFB6, EIF3L, FBXL19, KNG1, PRG2, TPI1, and UMOD, or the expression amount of a gene encoding the protein.

[0020] In one specific embodiment of the present invention, the agent for measuring the amount of the protein may be an antibody or aptamer that specifically binds to the protein.

[0021] In another specific embodiment of the present invention, the agent for measuring the expression level of the gene encoding the protein may be a primer set or probe set that specifically binds to the mRNA of the gene.

[0022] In addition, the present invention provides a kit for predicting treatment response to a biological agent in an asthma patient, comprising a composition and instructions for predicting treatment response to a biological agent in an asthma patient.

[0023] In addition, the present invention provides a diagnostic device for predicting the therapeutic response of an asthma patient to a biological agent, comprising a composition for predicting the therapeutic response of an asthma patient to a biological agent as an active ingredient.

[0024] In addition, the present invention provides a method for treating asthma, comprising the steps of: a) administering a pharmaceutically effective amount of a biological agent to an asthma patient; b) obtaining a biological sample from the asthma patient; c) measuring the amount of a protein expressed from one or more genes selected from the group consisting of BPIFB6, EIF3L, FBXL19, KNG1, PRG2, TPI1, and UMOD contained in the biological sample or the expression level of the gene; d) comparing the measured amount of the protein or the expression level of the gene with the amount of the same protein or the expression level of the gene in a control group; and e) when the measured amount of the protein or the expression level of the gene is significantly increased compared to the amount in the control group, administering the pharmaceutically effective amount of the biological agent again to the patient.

[0025] In addition, the present invention provides a method for predicting treatment response of an asthma patient to a biological agent, the method comprising, as an active ingredient, a preparation that measures the amount of a protein expressed from one or more genes selected from the group consisting of ACADL, APOC4, CCDC158, CDHR5, CLIP1, COL1A1, CTCFL, DNAI3, FAM98C, FGD2, GPT2, H2BK1, MDN1, MS4A1, NBL1, PIKFYVE, ROCK2, SRRM1, BPIFB6, EIF3L, FBXL19, KNG1, PRG2, TPI1, and UMOD, or the expression amount of a gene encoding the protein.

[0026] In addition, the present invention provides a composition comprising as an active ingredient an agent that measures the amount of a protein expressed from one or more genes selected from the group consisting of ACADL, APOC4, CCDC158, CDHR5, CLIP1, COL1A1, CTCFL, DNAI3, FAM98C, FGD2, GPT2, H2BK1, MDN1, MS4A1, NBL1, PIKFYVE, ROCK2, SRRM1, BPIFB6, EIF3L, FBXL19, KNG1, PRG2, TPI1, and UMOD, or the expression level of a gene encoding the protein, for use in predicting the therapeutic response to a biological agent in an asthma patient.

[0027] In addition, the present invention provides a use for preparing a formulation for predicting therapeutic responsiveness to a biological agent in an asthma patient, the formulation comprising as an active ingredient an agent that measures the amount of a protein expressed from one or more genes selected from the group consisting of ACADL, APOC4, CCDC158, CDHR5, CLIP1, COL1A1, CTCFL, DNAI3, FAM98C, FGD2, GPT2, H2BK1, MDN1, MS4A1, NBL1, PIKFYVE, ROCK2, SRRM1, BPIFB6, EIF3L, FBXL19, KNG1, PRG2, TPI1, and UMOD, or the expression level of a gene encoding the protein.

[0028] The method of providing information on predicting the therapeutic response of an asthma patient to a biological agent according to the present invention can identify a biological agent that exhibits a therapeutic effect suitable for each asthma patient, thereby significantly reducing the treatment cost and suffering of asthma patients, and can also significantly increase the therapeutic effect by selecting a biological agent that is suitable for the patient.

[0029] Figure 1 is a diagram showing the frequency of increased plasma proteins by variable in a group with a poor prognosis for asthma treatment according to one embodiment of the present invention.

[0030] Figure 2 is a diagram showing the frequency of increased plasma proteins by variable in a group with a good prognosis for asthma treatment according to one embodiment of the present invention.

[0031] The present invention comprises the steps of: a) measuring the amount of a protein contained in a biological sample obtained from an asthma patient administered a biological agent or the expression level of a gene encoding the protein; And b) a step of comparing the amount of the measured protein or the expression level of the gene encoding the protein with the amount of the protein or the expression level of the gene encoding the protein of a control group, wherein the protein is a protein expressed from one or more genes selected from the group consisting of ACADL, APOC4, CCDC158, CDHR5, CLIP1, COL1A1, CTCFL, DNAI3, FAM98C, FGD2, GPT2, H2BK1, MDN1, MS4A1, NBL1, PIKFYVE, ROCK2, SRRM1, BPIFB6, EIF3L, FBXL19, KNG1, PRG2, TPI1, and UMOD, and the control group is a blood sample obtained from the asthma patient before the administration of the biological agent. A method for providing information on predicting the treatment responsiveness of an asthma patient to a biological agent is provided.

[0032] In one specific embodiment of the present invention, the method may further include a step of classifying the therapeutic response to the biological agent as poor when the amount of a protein expressed from one or more genes selected from the group consisting of c) ACADL, APOC4, CCDC158, CDHR5, CLIP1, COL1A1, CTCFL, DNAI3, FAM98C, FGD2, GPT2, H2BK1, MDN1, MS4A1, NBL1, PIKFYVE, ROCK2, and SRRM1 or the expression level of the genes is significantly increased compared to the control group.

[0033] In another specific embodiment of the present invention, the method may further include the step of classifying the patient as exhibiting a therapeutic effect for the biological agent when the amount of protein expressed from one or more genes selected from the group consisting of c) BPIFB6, EIF3L, FBXL19, KNG1, PRG2, TPI1, and UMOD or the expression level of the genes is significantly increased compared to the control group.

[0034] The method of the present invention provides information on predicting treatment response of an asthma patient to a biological agent, and relates to a method for predicting or monitoring an asthma patient showing a treatment response to a biological agent by using a protein expressed from one or more genes selected from the group consisting of ACADL, APOC4, CCDC158, CDHR5, CLIP1, COL1A1, CTCFL, DNAI3, FAM98C, FGD2, GPT2, H2BK1, MDN1, MS4A1, NBL1, PIKFYVE, ROCK2, SRRM1, BPIFB6, EIF3L, FBXL19, KNG1, PRG2, TPI1, and UMOD contained in the blood protein of an asthma patient as a biomarker.

[0035] In the present specification, “a method for providing information on predicting treatment responsiveness to a biological agent in an asthma patient” may be used interchangeably with, but is not limited to, “a method for analyzing whether an asthma patient is highly sensitive to a biological agent” or “a method for characterizing a responder to a biological agent for treating asthma.”

[0036] Additionally, in the present specification, the “step of classifying a patient as having poor therapeutic responsiveness to a biological agent” may be used interchangeably with the “step of determining that an asthma patient is not susceptible to a biological agent,” and the “step of classifying a patient as having a therapeutic response to a biological agent” may be used interchangeably with the “step of determining that an asthma patient is susceptible to a biological agent,” but is not limited thereto.

[0037]

[0038] In this specification, “asthma” is a disease of the “bronchi,” the passages connecting to the lungs, which is a disease in which the bronchi are severely narrowed due to inflammation of the bronchi, causing repeated symptoms such as coughing, wheezing, shortness of breath, and chest tightness. Asthma includes, but is not limited to, allergic asthma or non-allergic asthma, severe asthma or uncontrolled asthma, adult asthma or childhood asthma. The “severe asthma” or “uncontrolled asthma” generally corresponds to, but is not limited to, severe asthma as defined in the American Thoracic Society and the European Respiratory Society (ATS / ERS) guidelines. The “uncontrolled asthma” refers to asthma in which the asthma condition is not controlled despite appropriate step-by-step asthma treatment, such as inhaled steroids appropriate for each asthma severity, and may include one or more of the following conditions, but is not limited to:

[0039] 1) Low symptom control: Partial control status according to the Korean Asthma Management Guidelines, or a score of less than 20 on the asthma control test.

[0040] 2) Frequent asthma exacerbations: Asthma exacerbations requiring systemic steroids for more than 3 days more than twice a year

[0041] 3) Severe exacerbation: When asthma worsens and hospitalization or mechanical ventilation is required.

[0042] 4) Airflow limitation: FEV1 < 80% after bronchodilator use.

[0043] In this specification, “biological agent” refers to a general term for a medical preparation made from a living organism, examples of which include serum, antigen, antibody, vitamin, hormone, cytokine, cell, plasma, etc., and preferably omalizumab, mepolizumab, reslizumab, benralizumab, dupilumab, but is not limited thereto if it is a biological preparation that can be used as a treatment for asthma. The dosage of the biological agent of the present invention may vary depending on various factors including the activity of the specific compound used, age, body weight, general health, sex, diet, administration time, administration route, excretion rate, drug combination, and the severity of the specific disease to be prevented or treated, and may be appropriately selected by those skilled in the art, although it may vary depending on the patient's condition, body weight, degree of disease, drug form, administration route, and period, and may be administered at 0.0001 to 500 mg / kg or 0.001 to 500 mg / kg per day. Administration may be administered once a day or divided into several times. The above dosage does not limit the scope of the present invention in any way. In addition, the biological agent may be obtained as a commercial product, or may be obtained by a method such as manufacturing by a known method.

[0044] In this specification, "biological sample" means any sample that can be obtained non-invasively and contains a protein or mRNA, and preferably includes, but is not limited to, blood, plasma, serum, bone marrow, tissue, cells, saliva, sputum, peritoneal fluid, hair, urine, feces, cerebrospinal fluid, various secretions, etc. The sample may be pretreated by methods such as homogenization, filtration, distillation, extraction, concentration, inactivation of interfering components, and addition of reagents before use for detection or diagnosis.

[0045] In this specification, “response” can be defined as “responsive” to a biological agent if asthma symptoms are prevented, improved, or treated as a result of biological agent treatment in an asthma patient. Prevention, improvement, or treatment of asthma can be measured in various known ways. The “prevention” refers to a broad concept of blocking the occurrence of asthma, and preferably includes both primary prevention, which prevents the occurrence before it occurs, and secondary prevention, which detects the occurrence early and treats it in a timely manner, but is not limited to the process and / or activity for dealing with the occurrence of asthma. The “treatment” refers to a broad concept of dealing with the occurrence of asthma, and is not limited to the process and / or activity for treating, curing, alleviating, or reducing asthma. The “improvement” refers to an act of at least reducing a parameter related to the condition being treated, for example, the severity of symptoms.

[0046] In this specification, the “method for providing information”, that is, the “method for providing information”, refers to a method for providing information on the asthma treatment effect of a biological agent, and means a method for obtaining or predicting information on the possibility that asthma symptoms will be prevented, improved, or treated when the biological agent is administered to an asthma patient. More specifically, it is a concept that includes all of selecting a biological agent suitable for an asthma patient, determining the susceptibility of an asthma patient to a biological agent, determining the prognosis of an asthma patient, or therametrics (e.g., monitoring the condition of an asthma patient to provide information on the treatment efficacy).

[0047] In addition, the present invention provides a kit for predicting treatment responsiveness to a biological agent in an asthma patient, comprising a composition and instructions for predicting treatment responsiveness to a biological agent in an asthma patient.

[0048] In the present specification, the “kit” of the present invention may be at least one selected from the group consisting of a microarray, an aptamer chip kit, an ELISA (Enzyme Linked Immunosorbent Assay) kit, a blotting kit, an immunoprecipitation kit, an immunofluorescence test kit, a protein chip kit, a reverse transcription polymerase chain reaction (RT-PCR) kit, and a real-time polymerase chain reaction (qRT-PCR) kit, but is not limited thereto.

[0049] In the present specification, the term "kit" means a diagnostic device capable of predicting the therapeutic responsiveness of an asthma patient to a biological agent by measuring the amount of a protein expressed from one or more genes selected from the group consisting of ACADL, APOC4, CCDC158, CDHR5, CLIP1, COL1A1, CTCFL, DNAI3, FAM98C, FGD2, GPT2, H2BK1, MDN1, MS4A1, NBL1, PIKFYVE, ROCK2, SRRM1, BPIFB6, EIF3L, FBXL19, KNG1, PRG2, TPI1, and UMOD contained in a biological sample, preferably blood, or the expression amount of a gene encoding the protein, and there is no limitation as long as it is in a form capable of measuring the amount of the protein from a blood sample isolated from an asthma patient.

[0050] In another specific embodiment of the present invention, the step of measuring the amount of the protein or the expression level of the gene encoding the protein may be performed by Western blotting, enzyme linked immunosorbent assay (ELISA), radioimmunoassay, radioimmunodiffusion, Ouchterlony immunodiffusion, Rocket immunoelectrophoresis, immunohistochemical staining, immunoprecipitation assay, complete fixation assay, flow cytometry (FACS), protein chip, gas chromatography, liquid chromatography, mass spectrometry, measurement using affibody, proximity extension assay, reverse transcription polymerase chain reaction (RT-PCR), competitive RT-PCR, real-time It can be measured by one or more methods selected from the group consisting of, but not limited to, real-time RT-PCR, RNase protection assay, Northern blotting, and DNA microarray chip.

[0051] In the present specification, the “instruction” of the present invention may be, but is not limited to, classifying a treatment response to a biological agent as poor when the amount of a protein expressed from one or more genes selected from the group consisting of ACADL, APOC4, CCDC158, CDHR5, CLIP1, COL1A1, CTCFL, DNAI3, FAM98C, FGD2, GPT2, H2BK1, MDN1, MS4A1, NBL1, PIKFYVE, ROCK2, and SRRM1 or the expression level of the genes is significantly increased compared to the control group.

[0052] In the present specification, the “instruction” of the present invention may be, but is not limited to, classifying a patient as exhibiting a therapeutic effect for a biological agent when the amount of protein expressed from one or more genes selected from the group consisting of BPIFB6, EIF3L, FBXL19, KNG1, PRG2, TPI1, and UMOD or the expression amount of the genes is significantly increased compared to a control group.

[0053] In this specification, the term "diagnosis instrument" means a device capable of measuring the therapeutic response to a biological agent in vitro based on a biological sample generated from a human body, such as sputum, blood, plasma, saliva, urine, etc., preferably blood, and preferably may include an inlet for adding a biological sample, a body capable of measuring the amount of a protein expressed from one or more genes selected from the group consisting of ACADL, APOC4, CCDC158, CDHR5, CLIP1, COL1A1, CTCFL, DNAI3, FAM98C, FGD2, GPT2, H2BK1, MDN1, MS4A1, NBL1, PIKFYVE, ROCK2, SRRM1, BPIFB6, EIF3L, FBXL19, KNG1, PRG2, TPI1, and UMOD or the expression amount of a gene encoding the protein, an output unit for showing the measured result, etc., but there are no limitations as long as the device is of a type capable of measuring the amount of the protein contained in a biological sample. does not exist.

[0054] In addition, the present invention provides a method for treating asthma, comprising the steps of: a) administering a pharmaceutically effective amount of a biological agent to an asthma patient;

[0055] b) obtaining a biological sample from the asthma patient;

[0056] c) a step of measuring the amount of protein expressed from one or more genes selected from the group consisting of BPIFB6, EIF3L, FBXL19, KNG1, PRG2, TPI1, and UMOD contained in the biological sample or the expression level of the gene;

[0057] d) a step of measuring the amount of protein expressed from one or more genes selected from the group consisting of BPIFB6, EIF3L, FBXL19, KNG1, PRG2, TPI1, and UMOD contained in a biological sample obtained from a control group, or the expression level of the gene, and comparing the amount of protein or the expression level of the gene measured in step c); and

[0058] e) Provided is a method for treating asthma, comprising a step of re-administering a pharmaceutically effective amount of a biological agent to the patient when the amount of the measured protein or the expression amount of the gene is significantly increased compared to the amount of the control group.

[0059] In addition, the present invention provides a method for predicting treatment response of an asthma patient to a biological agent, the method comprising, as an active ingredient, a preparation that measures the amount of a protein expressed from one or more genes selected from the group consisting of ACADL, APOC4, CCDC158, CDHR5, CLIP1, COL1A1, CTCFL, DNAI3, FAM98C, FGD2, GPT2, H2BK1, MDN1, MS4A1, NBL1, PIKFYVE, ROCK2, SRRM1, BPIFB6, EIF3L, FBXL19, KNG1, PRG2, TPI1, and UMOD, or the expression amount of a gene encoding the protein.

[0060] In addition, the present invention provides a composition comprising as an active ingredient an agent that measures the amount of a protein expressed from one or more genes selected from the group consisting of ACADL, APOC4, CCDC158, CDHR5, CLIP1, COL1A1, CTCFL, DNAI3, FAM98C, FGD2, GPT2, H2BK1, MDN1, MS4A1, NBL1, PIKFYVE, ROCK2, SRRM1, BPIFB6, EIF3L, FBXL19, KNG1, PRG2, TPI1, and UMOD, or the expression level of a gene encoding the protein, for use in predicting the therapeutic response to a biological agent in an asthma patient.

[0061] In addition, the present invention provides a use for preparing a formulation for predicting therapeutic responsiveness to a biological agent in an asthma patient, the formulation comprising as an active ingredient an agent that measures the amount of a protein expressed from one or more genes selected from the group consisting of ACADL, APOC4, CCDC158, CDHR5, CLIP1, COL1A1, CTCFL, DNAI3, FAM98C, FGD2, GPT2, H2BK1, MDN1, MS4A1, NBL1, PIKFYVE, ROCK2, SRRM1, BPIFB6, EIF3L, FBXL19, KNG1, PRG2, TPI1, and UMOD, or the expression level of a gene encoding the protein.

[0062] Hereinafter, preferred examples are presented to aid in understanding the present invention. However, the following examples are provided solely to facilitate a better understanding of the present invention, and the scope of the present invention is not limited by the following examples.

[0063]

[0064] [Example]

[0065] Example 1: Blood proteomic analysis

[0066] To identify blood biomarkers for predicting the asthma treatment effect of biological agents, blood samples were obtained before treatment and at 1 or 6 months after a single treatment from 83 patients (PRISM study cohort hosted by the Ministry of Science and Technology) who visited for asthma treatment. Asthma was treated with biological agents such as mepolizumab, dupilumab, reslizumab, or omalizumab, and the patients were classified into responder, excellent responder (excel_resp), super-responder (super_res), ex-50per, ACT-3more, or FEV1ml_more10p100ml groups according to asthma treatment prognostic variables. responder is a response prognostic variable that indicates a 50% or greater decrease in annual exacerbation rate or a 50% or greater decrease in the mean daily oral corticosteroid dose (mOCS), super_res is a response prognostic variable that indicates zero annual exacerbation rate or mOCS, excel_resp is a response prognostic variable that indicates zero clinical asthma exacerbations (CAEs) for 6 months, ex-50per is a response prognostic variable that indicates a 50% or greater decrease in CAEs, ACT-3more is a response prognostic variable that indicates a 3-point or greater increase in the Asthma Control Test (ACT) score, and FEV1ml_more10p100ml is a response prognostic variable that indicates 100 mL or greater or a 10% or greater increase in prebronchodilator FEV1. If the criteria were met, it was classified as 1=Yes, indicating treatment responsiveness. If the criteria were not met, it was classified as 0=No, indicating treatment responsiveness. Furthermore, blood proteomes before and after treatment were compared and analyzed according to each response prognostic variable.More specifically, to first obtain the proteome, the obtained blood sample was centrifuged at 1,000xg for 10 minutes to isolate only the supernatant, i.e., plasma, and the amount of protein contained in the supernatant was quantified using Pierce™BCA Protein Assay Kit (Thermo Fisher SCIENTIFIC). Then, 300μg of the obtained protein was used for S-Trap™ (ProtiFi, LLC)-based peptide digestion under 5% SDS conditions, and quantitative proteome analysis was performed by LC-MS using Thermo Q Exactive HFx equipped with NanoLC. As a result, a total of 288 blood proteins were identified, and in order to identify proteins that appeared in the patient groups divided according to the treatment effect, proteins whose protein amount was increased by 1.5 times or more in each group and whose p-value was less than 0.05 were identified.

[0067] In the responder group, 8 proteins were identified that were increased by more than 1.5 times and had a p-value of less than 0.05, and are shown in Table 1. As shown in Table 1, PIKFYVE, CCDC158, CTCFL, GPT2, CDHR5, FAM98C, NBL1, and APOC4 were confirmed to be increased in the group with poor treatment prognosis.

[0068]

[0069]

[0070] In the super_res group, five proteins were identified as having a p-value less than 0.05 and an increase of more than 1.5 times, and are shown in Table 2. As shown in Table 2, SRRM1, MS4A1, CCDC158, and DNAI3 were found to be increased in the group with a poor prognosis, while KNG1 was found to be increased in the group with a good prognosis.

[0071]

[0072]

[0073] In the excel_resp group, 11 proteins were identified that were increased by more than 1.5 times and had a p-value of less than 0.05, and are shown in Table 3. As shown in Table 3, PIKFYVE, SRRM1, MS4A1, CCDC158, ROCK2, DNAI3, CLIP1, H2BK1, ACADL, APOC4, and CTCFL were all confirmed to be increased in the group with poor treatment prognosis.

[0074]

[0075]

[0076] In the ex-50per group, 10 proteins were identified that were increased by more than 1.5 times and had a p-value of less than 0.05, and are shown in Table 4. As shown in Table 4, SRRM1, ROCK2, ACADL, H2BK1, CLIP1, NBL1, COL1A1, CDHR5, and APOC4 were increased in the group with poor treatment prognosis, and TPI1 was confirmed to be increased in the group with good prognosis.

[0077]

[0078]

[0079] In the ACT-3more group, five proteins were identified as having a p-value less than 0.05 and an increase of more than 1.5 times, and are shown in Table 5. As shown in Table 5, FGD2 and MDN1 were found to be increased in the group with a poor prognosis, while EIF3L, PRG2, and FBXL19 were found to be increased in the group with a good prognosis.

[0080]

[0081]

[0082] In the FEV1ml_more10p100ml group, three proteins were identified that increased by more than 1.5 times and had a p-value of less than 0.05, and these are shown in Table 6. As shown in Table 6, PRG2, BPIFB6, and UMOD were confirmed to be increased in the group with a good prognosis.

[0083]

[0084]

[0085] The results of analyzing the frequency of increased plasma proteins by variable in the group with poor asthma treatment prognosis are shown in Figure 1.

[0086] As shown in Fig. 1, the proteins that were commonly increased in two or more variables were APOC4, SRRM1, CCDC158, ROCK2, CLIP1, H2BK1, ACADL, PIKFYVE, CTCFL, MS4A1, DNAI3, NBL1, and CDHR5, a total of 13 proteins. Through this, it was confirmed that the above proteins can be used as biomarkers to predict or monitor patients who show a negative prognosis to treatment with biological agents.

[0087] And the results of analyzing the frequency of increased proteins by variable in the group with good asthma treatment prognosis are shown in Figure 2.

[0088] As shown in Fig. 2, the proteins that were commonly increased in two or more variables were PRG2, a total of one protein, and through this, it was confirmed that the above proteins can be used as biomarkers to predict or monitor patients showing a positive prognosis in response to treatment with biological agents.

[0089]

[0090] The foregoing description of the present invention is provided for illustrative purposes only. Those skilled in the art will readily appreciate that the present invention can be readily modified into other specific forms without altering the technical spirit or essential characteristics of the present invention. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive.

[0091] The method of providing information on predicting the therapeutic response of an asthma patient to a biological agent according to the present invention can identify a biological agent that exhibits a therapeutic effect suitable for each asthma patient, thereby significantly reducing the treatment cost and suffering of asthma patients, and can also significantly increase the therapeutic effect by selecting a biological agent that is suitable for the patient, and thus has industrial applicability.

Claims

1. a) a step of measuring the amount of a protein contained in a biological sample obtained from an asthma patient administered a biological agent or the expression level of a gene encoding the protein; and b) A method for providing information on predicting treatment responsiveness to a biological agent of an asthma patient, comprising a step of comparing the amount of the measured protein or the expression level of the gene encoding the protein with the amount of the protein or the expression level of the gene encoding the protein of a control group, The above protein is a protein expressed from one or more genes selected from the group consisting of ACADL, APOC4, CCDC158, CDHR5, CLIP1, COL1A1, CTCFL, DNAI3, FAM98C, FGD2, GPT2, H2BK1, MDN1, MS4A1, NBL1, PIKFYVE, ROCK2, SRRM1, BPIFB6, EIF3L, FBXL19, KNG1, PRG2, TPI1, and UMOD. The above control group is a biological sample obtained from an asthma patient before administration of the biological agent.

2. In paragraph 1, The method is characterized in that it further comprises a step of classifying the treatment response to the biological agent as poor when the amount of a protein expressed from one or more genes selected from the group consisting of c) ACADL, APOC4, CCDC158, CDHR5, CLIP1, COL1A1, CTCFL, DNAI3, FAM98C, FGD2, GPT2, H2BK1, MDN1, MS4A1, NBL1, PIKFYVE, ROCK2, and SRRM1 or the expression amount of a gene encoding the protein is significantly increased compared to the control group.

3. In either paragraph 1 or paragraph 2, The method is characterized in that it further comprises a step of classifying the patient as exhibiting a therapeutic effect for a biological agent when the amount of a protein expressed from one or more genes selected from the group consisting of c) BPIFB6, EIF3L, FBXL19, KNG1, PRG2, TPI1, and UMOD or the expression amount of a gene encoding the protein is significantly increased compared to a control group.

4. In any one of paragraphs 1 to 3, A method, characterized in that the biological sample is blood, plasma or serum.

5. In any one of paragraphs 1 to 4, A method, characterized in that the biological agent is at least one selected from the group consisting of omalizumab, mepolizumab, reslizumab, benralizumab, and dupilumab.

6. In any one of paragraphs 1 to 5, The steps for measuring the amount of the protein or the expression level of the gene encoding the protein include western blotting, enzyme linked immunosorbent assay (ELISA), radioimmunoassay, radioimmunodiffusion, Ouchterlony immunodiffusion, Rocket immunoelectrophoresis, immunohistochemical staining, immunoprecipitation assay, complete fixation assay, flow cytometry (FACS), protein chip, gas chromatography, liquid chromatography, mass spectrometry, measurement using affibody, proximity extension assay, reverse transcription polymerase chain reaction (RT-PCR), competitive RT-PCR, and real-time reverse transcription. A method characterized in that the measurement is performed by at least one method selected from the group consisting of polymerase chain reaction (Real-time RT-PCR), RNase protection assay, Northern blotting, and DNA microarray chip.

7. A composition for predicting the therapeutic response of an asthma patient to a biological agent, comprising as an active ingredient an agent that measures the amount of a protein expressed from one or more genes selected from the group consisting of ACADL, APOC4, CCDC158, CDHR5, CLIP1, COL1A1, CTCFL, DNAI3, FAM98C, FGD2, GPT2, H2BK1, MDN1, MS4A1, NBL1, PIKFYVE, ROCK2, SRRM1, BPIFB6, EIF3L, FBXL19, KNG1, PRG2, TPI1, and UMOD, or the expression amount of a gene encoding the protein.

8. In paragraph 7, A composition characterized in that the agent for measuring the amount of the protein is an antibody or aptamer that specifically binds to the protein.

9. In either of paragraphs 7 or 8, A composition characterized in that the preparation for measuring the expression level of the gene encoding the above protein is a primer set or probe set that specifically binds to the mRNA of the above gene.

10. A kit for predicting treatment response to a biological agent in an asthma patient, comprising a composition for predicting treatment response to a biological agent in any one of claims 7 to 9, and an instruction manual.

11. A diagnostic device for predicting the therapeutic response of an asthma patient to a biological agent, comprising as an active ingredient a composition for predicting the therapeutic response of an asthma patient to a biological agent according to any one of claims 7 to 9. 12.a) Administering a pharmaceutically effective amount of a biological agent to an asthma patient; b) obtaining a biological sample from the asthma patient; c) a step of measuring the amount of protein expressed from one or more genes selected from the group consisting of BPIFB6, EIF3L, FBXL19, KNG1, PRG2, TPI1, and UMOD contained in the biological sample or the expression level of the gene; d) a step of comparing the amount of the measured protein or the expression level of the gene with the amount of the same protein or the expression level of the gene of the control group; and e) A method for treating asthma, comprising a step of re-administering a pharmaceutically effective amount of a biological agent to the patient when the amount of the measured protein or the expression amount of the gene is significantly increased compared to the amount of the control group.

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